본 연구는 인체 및 온혈 동물의 기회감염 병원세균으로 알려진 Pseudomonas oryzihabitans에 의한 양파 구에 발생한 중심 흑화병의 첫 보고이다. 본 연구의 목적은 양파 구 흑화병을 일으키는 원인 병원체를 분리 및 동정하고 미생물학적 특성을 조사하는데 있다. 2021년 국내 양파 구에서 소비자 혐오감을 유발할 수 있는 세균 흑화병이 관찰되었다. 흑갈색 색소가 침착된 조직과 흑색 색소 과육 조직에서 그람음성, 비포자형성, 막대형 pseudomonad들이 분리되었다. 병원성 검정을 통해 형성된 인공 감염후 발생한 증상은 자연 감염과 같았고 멸균 증류수를 처리한 음성 대조구에서는 흑화병이 발견되지 않았다. 병원성 검정, API (analytical profile index) 시험, MALDI-TOF MS (Whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry), 16S rRNA 유전자 영역 염기서열분석 및 유전적 유연관계분석을 통해 원인 병원체를 Pseudomonas oryzihabitans로 동정하였다. 대규모 포장 또는 수확후병으로 인한 양파 피해사례가 보고된 적은 없지만, 본 연구에서 얻은 결과는 P. oryzihabitans가 잠재적으로 양파의 글로벌 생산 및 운송과 식품 안전을 위협할 수 있음을 시사한다. 또한 인체 기회감염 병원체이기 때문에 양파로부터 언제든지 인체로 감염될 수 있는 위험성을 갖고 있다고 볼 수 있다.

The bacterial soft-rot disease is one of the most critical diseases in vegetables such as Chinese cabbage. The researchers isolated two bacteria (Pseudomonas kribbensis and Pantoea vagans) from diseased tissue samples of Chinese cabbages and confirmed them as being the strains that cause soft-rot disease. Lactic-acid bacteria (LAB), were screened and used to control soft-rot disease bacteria. The researchers tested the treatments with hypochlorous acid water (HAW) and LAB supernatant to control soft-rot disease bacteria. The tests confirmed that treatments with the HAW (over 120 ppm) or LAB (Lactobacillus plantarum PL203) culture supernatants (0.5 mL) completely controlled both P. kribbensis and P. vagans.

A gram-positive bacterium was isolated from the spent substrate of Agaricus bisporus that showed a marked antagonistic activity against Pseudomonas agarici. It was identified as Bacillus safensis HC42 based on its cultural, biochemical, and physiological characteristics, and 16S rRNA sequence. B. safensis HC42 was saprophytic, but not parasitic or pathogenic, in cultivated mushrooms and showed strong inhibition of P. agarici in vitro. Moreover, it showed a control efficacy of 66 % against browning disease caused by P. agarici in Agaricus bisporus. Therefore, B. safensis HC42 may be useful in the future for the development of a biocontrol system.

The aim of this study was to isolate and identify marine bacterium with anti-methicillinresistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were 25℃ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a C18 ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of 250 μg mL-1 concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.

We experimented to see if the tolerance of Agaricus bisporus to Pseudomonas sp. causing the bacterial brown blotch disease which is causing great damage in the button mushroom cultivation was inherited. There was no correlation between mycelial growth rate and mushroom tolerance to pathogens at each temperature. In the mycelial stage, the strains tolerance to Pseudomonas sp. were not as strong as those of their parents, but they were generally stronger or more tolerance than those of weaker strain (ASI1321). In fruiting body, tolerance was decreased compared to mycelial. In fruiting body, later generations of two strains had similar tolerance, unlike strong or weak tolerance of parental generation. Therefore, browning of fruit body is thought to be caused not only by tolerance but also by various other factors. Especially, the future generation of the strains which were tolerance to Pseudomonas sp. were very weak in the mycelial stage, indicating that the tolerance of the parental generation was inherited to the later generations. The damage of each pathogen was different in mycelium and fruiting body. P. tolaasii caused higher browning than P. agarici in fruit body. P. reactans did not have a significant effect on the mycelium but affected the browning of the fruit bodies. P. agarici had higher mycelial growth inhibitory ability than fruiting body.

곤충병원균인 Pseudomonas entomophila를 Riptortus pedestris의 체내에 주사하였을 때, 곤충을 사멸시키는 효과를 나타내는 물질을 찾기 위하여, P. entomophila를 배양한 배양액을 이온교환 크로마토그래피 및 크기배제 크로마토그래 피를 통해 살충 효과를 나타내는 물질을 정제 하였다. 그 결과 R. pedestris에 대해 살충효과를 보이는 독성 물질은 metalloproteinase의 일종인 AprA 단백질인 것으로 나타났다. 정제 된 AprA를 R. pedestris에 주사하였을 때, 곤충이 매우 빠른 속도로 사멸하는 양상을 보였다. AprA 단백질이 어떠한 작용 기전으로 R. pedestris에 독성 효과를 보이는지 알기 위하여, 정제 된 AprA를 R. pedestris의 체내로 주사한 후 hemocyte (혈구)의 숫자를 조사한 결과, AprA를 주사 한 직후에는 혈구의 숫자가 증가하였으나, 1시간 이후부터는 혈구의 수가 급격히 감소하기 시작하였고, 결국은 모든 곤충이 사멸하였다. 본 연구진은 AprA가 R. pedestris의 면역반응을 어떻게 억제하는지 알기 위해, AprA가 R. pedestris의 혈구를 제거하는 기작을 보고하고자 한다.

This study was conducted to investigate optimum conditions for mass production of ntagonistic microbes Alcaligenes sp. HC12. Alcaligenes sp. HC12 had a potent biological control agent to control browning disease caused by Pseudomonas agarici. Alcaligenes sp. HC12 markedly showed the antagonistic activity against Pseudomonas agarici, the most destructive pathogen of cultivated mushrooms. To define the optimum conditions for the mass production of the Alcaligenes sp. HC12, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 9.0 and 30o, respectively. The optimal concentration of medium elements for the growth of pathogen inhibitor bacterium(Alcaligenes sp. HC12) was determined as follows: 0.5% dextrine, 1.5% yest extract, 1.0% NaNO3, 0.5% KH2PO4, and 1.5% asparagine.

버섯 세균갈성색무늬병원균인 Pseudomonas tolaasii에 대한 길항미생물로 보고된 Pseudomonas azotoformans HC5 균주의 배양적 특성과 대량 배양을 위한 최적배양조건을 설정하였다. HC5 균주의 생육온도는 10~20oC, pH는 6.0~9.0의 범위에서 왕성한 생육을 보였다. 대량배양을 위한 효율적인 영양원 선발을 위하여 기본배지에 탄소원 fructose 등 18종, 무기질소원 NH4Cl 등 6종, 유기질소원 peptone 등 6종 그리고 아미노산 asparagine 등 11종을 각각 1%씩 첨가하였고, 무기염류 13종을 1 mM 농도로 첨가하여 각각에 대한 생육에 미치는 영향을 조사하였다. 또한 선발된 각각의 영양성분들에 대한 최적 농도를 조사하기 위하여 각각의 성분을 최소 0.1%에서 최대 4.0%까지 배지에 첨가하여 배양 후 생육정도를 조사하였다. 그 결과, 대량배양을 위한 생육최적조건은 온도 15oC, pH 6, 탄소원 0.6% adonitol, 유기질소원 1.5% yeast extract, 무기질소원 0.8% NH4H2PO4,아미노산 0.2% asparagine 그리고 무기염류는 5 mM MgSO4에서 왕성한 생육을 보였다.

Sesame is a major cooking oil crop in Korea. One of the primary problems in sesame cultivation is low healthy stand establishment due to the occurrence of seedling rot and damping-off resulting from a complex of soil-borne pathogens in the field. To address the problem, the bioformulation of Pseudomonas fluorescens M45 was prepared in powder form using clay and vermiculite, and was evaluated for its effect on biological control of soil-borne pathogens in sesame cultivation. In the petri dish trial, the emergence rate was overall good (> 92%) regardless of seeds being pelleted and/or M45-treated. In both pot and field trials containing disease-conducive soils, seed-pelleting substantially reduced emergence rate, whereas seed-pelleting with M45 significantly improved the emergence rate (> 26%). The emergence rate of sesame seeds treated with the strain M45 was greater than 30% regardless of seed pelletization. We also found that M45r colonized in the roots at the density of 1.6×105 cfu/g. With aid of the bioformulation, however, root colonization of the strain was significantly increased to 4.0×106 cfu/g. The powder formulation with strain M45 enhanced the rate of healthy stand establishment in disease-conducive soil. Therefore, bioformulation with strain M45 is a promising method to overcome problems associated with the successive cultivation of sesame.

본 연구는 P. aeruginosa와 E. hirae을 대상으로 orthophenylphenol 20%를 함유한 훈증소독제, Fumagari OPP®의 살균효과를 평가하기 위해 수행되었다. 예비 시험에서, P. aeruginosa와 E. hirae의 현탁액 균수는 각각 2.8 × 108 와 3.6 × 108CFU/mL이었으며, 모든 훈증소독제에 노출시킨 담체의 균수는 모두 평판배지법과 여과법으로 배양한 시험균주 현탁액의 균수의 50%보다 많았다. 또한, 대조 담체로부터 회복된 P. aeruginosa와 E. hirae 균수는 각각 2.9 × 106와 2.7 × 106CFU/mL이었다. 훈증소독제의 살균효과 시험에서는, 훈증소독제를 처리한 담체의 P. aeruginosa 와 E. hirae의 감소 균수는 각각 6.46와 5.19 logCFU/mL 로 나타났다. 이상의 결과로부터, 훈증소독제, Fumagari OPP®는 P. aeruginosa와 E. hirae에 대해 효과적인 살균력 을 갖는 것으로 확인되었으며, 병원성 세균에 오염된 식품재료 및 주방용품의 소독에 적용할 수 있을 것으로 사료된다.

The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 10,996 M/T in 2012. Several bacteria are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Pseudomonas agarici is the causal agent of browning disease of commercial mushrooms. Colonization of mushroom caps by the bacterium results in development of browning lesions on pileus. These lesions are superficial brown spots and can be round or spreading. But P. agarici never caused sunken lesions and rotting of the mushroom tissues. A Gram-positive bacterium was isolated from mushroom media that markedly showed the antagonistic activity against Pseudomonas agarici, the most destructive pathogen of cultivated mushrooms. The HC42 strain was selected as antagonistic bacterium by inhibition zone method and it was identified as Bacillus safensis. by the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA.. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for P. agarici cell, was sufficient for inhibition in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Agaricus bisporus. Control efficacy of browning disease of strain HC42 treatment was 66% on Agaricus bisporus. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 1.5% D-galactose, 1.5% yest extract, 1% NH4Cl, 1.5% KCl, and 1.0% L-asparagin at pH 6.0 at 25℃. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by P. agarici.

This test was performed to evaluate the bactericidal efficacy of a fumigation disinfectant containing 20% ortho-phenylphenol against Pseudomonas aeruginosa (P. aeruginosa) and Enterococcus hirae (E. hirae). In preliminary tests, P. aeruginosa and E. hirae working culture suspension number (N value) were 2.8 × 108 and 4.0 × 108CFU/mL, respectively. And all the colony numbers on the carriers exposed to the fumigant (n1, n2, n3) were higher than 0.5N1 (the number of bacterial test suspentions by pour plate method), 0.5N2 (the number of bacterial test suspentions by filter membrane method) and 0.5N1, respectively. In addition, the mean number of P. aeruginosa and E. hirae recovered on the control-carriers (T value) was 2.8 × 108 and 3.4 × 106 CFU/mL, respectively. In the bactericidal effect of the fumigant, the reduction number of 2.8 × 108 (d value) was 6.46 and 5.19 logCFU/mL, respectively. According to the French standard for the fumigant, the d value for the effective bactericidal fumigant should be over than 5 logCFU/mL. With the results from this study, the fumigation disinfectant containing 20% ortho-phenylphenol has an effective bactericidal activity, then the fumigant can be applied to disinfect food materials and kitchen appliances contaminated with the pathogenic bacteria.

버섯 세균갈성색무늬병원균인 Pseudomonas tolaasii에대한 독소저해균으로 보고된 Pseudomonas sp. HC1 균주의 배양적 특성과 최적 배양을 위한 대량배지를 선발하였다. CH1균주의 생육온도는 20~40oC, pH는 5.0~11.0까지넓은 범위에서 왕성한 생육을 보였다. 대량배양을 위한효율적인 영양원 선발을 위하여 기본배지에 탄소원fructose 등 18종, 무기질소원 NH4Cl 등 6종, 유기질소원peptone 등 6종 그리고 아미노산 asparagine 등 11종을 각각 1%씩 첨가하였고, 무기염류 13종을 1mM 농도로 첨가하여 각각에 대한 생육에 미치는 영향을 조사하였다.또한 선발된 각각의 영양성분들에 대한 최적 농도를 조사하기 위하여 각각의 성분을 0.1%에서 4.0%까지 배지에첨가하여 배양후 생육정도를 조사하였다. 그 결과, 대량배양을 위한 생육최적조건은 온도 20oC, pH5, 탄소원 0.9%dextrine, 유기질소원 1.5% yeast extract, 무기질소원0.5% (NH4)2HPO4, 아미노산 3.0% cysteine, 무기염류4mM FeCl3에서 왕성한 생육을 보였다.

목적: 황색포도상구균(Staphylococcus aureus)과 녹농균(Pseudomonas aeruginosa)으로 오염시킨 RGP 렌즈를 3가지 종류의 다목적용액에 침윤시킨 후 각 시간대별 두 균주의 항균효과를 비교하여 렌즈 소독에 필요한 침윤시간을 제시하고자 한다. 대상과 방법: 황색포도상구균과 녹농균의 배양액(농도 109 Colony Forming Unit/ml) 0.1ml와 LB broth 0.9ml 혼합액에 RGP 렌즈를 넣은 후 24시간 배양 후 실험을 시행하였다. 3가지 종류의 다목적용액을 1시간, 2시간, 3시간, 4시간, 5시간, 6시간, 7시간 동안 침윤시켜 항균 효과를 비교하였다. 결과: 황색포도상구균과 녹농균에 대한 다목적용액 간에 항균효과 차이는, 시간대별로 1시간 침윤으로 전체 균의 cfu가 약 43~47%, 55~63% 감소하였고, 2시간 침윤으로는 약 77~80%, 77~86% 감소하였다. 3시간 침윤으로는 약 97~98%, 85~97% cfu가 감소하였고, 4시간 침윤으로는 100%, 95~100% cfu가 감소하였다. 그리고 5시간, 6시간, 7시간 침윤으로는 다목적용액 모두 황색포도상구균(Staphylococcus aureus)과 녹농균(Pseudomonas aeruginosa)이 검출되지 않았다. 결론: 황색포도상구균은 다목적용액 4시간 이상의 침윤시간에서 검출되지 않았고 녹농균은 다목적용액 5시간 이상의 침윤시간에서 세균이 검출되지 않았다. 따라서 본 연구에 사용된 다목적용액을 RGP 콘택트렌즈 소독에 사용하는 경우 5시간 이상 침윤시켜야 할 것으로 사료된다.

곤충의 장내세균은 곤충 내에서 곤충의 소화, 발육, 번식, 생존 등 여러가지 역할 을 하는 것으로 밝혀져 있다. 특히, 여러 물질을 분해하는 물질을 분비하거나 항생물 질을 분비하기도 한다. 콩풍뎅이(Popillia mutans)에서 Pseudomonas aeruginosa외 에 4종의 균주가 확인되었으며 이 균주들을 Alternaria solani, Botrytis cinera, Colletotrichum gloeosporioides, Fusarium oxysporum, Phytophthora capsici, Rhizoctonia solani 6종의 병원균과 함께 PDA배지에서 대치배양하여 항균활성을 본 결과, Ps. aeruginosa균주가 A. solani, B. cinera, C. gloeosporioides, F. oxysporum, P. capsici 5종의 병원균 모두에 항균활성을 갖는 것으로 나타났다. 특히, B. cinera 와 P. capsici에 대해 비교적 높은 항균활성을 갖는 것을 볼 수 있었다.

Pseudomonas fluorescens B16 has been shown to be a promising biological control agent for the promotion of plant growth, root colonization, and suppression of bacterial wilt. This study investigated the dispersion of B16 from seed to radicle, and to the whole root system, and illustrated population changes within cucumber seeds and radicles using scanning electron microscopy (SEM), fluorescence microscopy (FM) and immunofluorescence microscopy (IFM). When cucumber seeds were soaked in a bacterial suspension of B16, bacterial cells entered the seeds through the pore at the base, colonized the inner plane, and proliferated germination advanced. Population densities of B16 were gradually increased, and increased initial populations within newly growing roots. The root surface was covered by cylindrical rod-shaped bacterial cells, and junctions between epidermal cells were covered preferentially. Previous studies exploring root colonization of rhizobacteria have not focused on colonized populations inside seeds. Our microscopic observations confirmed the importance of bacterial cells colonized inside seed coats. Population densities of B16 were then naturally sustained up to harvesting time. The results obtained in this study provide about a novel insight into the commercial application of biological control agents within seeds.