• Journal of Food Hygiene and Safety
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• v.16 no.3
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• pp.232-240
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• 2001

One hundred twenty five bacterial isolates were obtained from the brown blotch-diseased oyster mushrooms collected from markets. Among them, 45 were determined as pathogenic bacteria and white line forming organisms(WLFO) were 6 strains and white line reaction organisms (WLRO) were 6 strains. All of the white line forming isolates were identified as Pseudomonas tolaasii which is a known pathogen of brown blotch disease of oyster mushroom by GC-MIS(Gas chromatography-microbial identification system). Six of the white line reacting organisms were identified as P. chlomraphis, P. fluorescens biotype A and type C. The rest of them were P gingeri, P. agarici, P. fluorescens biotype B, P. chloroyaphis, non-pathogenic P. tolaasii, P. putida biotype A and B etc. For spectrum of activity of tolaasin, culture filtrates from pathogenic isolates were examined by browning of mushroom tissue and pitting of mushroom caps. The weak pathogenic bacteria didn't induce browning or pitting of mushroom tissue. On the other hand, strong pathogenic isolates showed browning and pitting reaction on mushroom. An extracellular toxin produced by P. tolaasii, was investigated. The hemolysis activity test of 6 strains identified as P. tolaasii were 0.8∼0.9 at 600 nm and 3 strains of WLRO were 0.9∼1.0 and Pseudomonas app. were 1.0∼1.2. Observation of fresh mushroom tissue using confocal laser scanning microscopy was carried out for images of optical sectioning and vertical sectioning. Also images of brown blotch diseased oyster mushroom tissue after contamination P. tolaasii was obtained by CLSM.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1087-1093
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• 2005

The population diversity and seasonal changes of bacterial communities in rice soils were monitored using both culture-dependent approaches and molecular methods. The rice field plot consisted of twelve subplots planted with two genetically-modified (GM) rice and two non-GM rice plants in three replicates. The DGGE analysis revealed that the bacterial community structures of the twelve subplot soils were quite similar to each other in a given month, indicating that there were no significant differences in the structure of the soil microbial populations between GM rice and non-GM rice during the experiment. However, the DGGE profiles of June soil after a sudden flooding were quite different from those of the other months. The June profiles exhibited a few intense DNA bands, compared with the others, indicating that flooding of rice field stimulated selective growth of some indigenous microorganisms. Phylogenetic analysis of l6S rDNA sequences from cultivated isolates showed that, while the isolates obtained from April soil before flooding were relatively evenly distributed among diverse genera such as Arthrobacter, Streptomyces, Terrabacter, and Bacillus/Paenibacillus, those from June soil after flooding mostly belonged to the Arthrobacter species. Phylogenetic analysis of 16S rDNA sequences obtained from the soil by cloning showed that April, August, and October had more diverse microorganisms than June. The results of this study indicated that flooding of rice fields gave a significant impact on the indigenous microbial community structure; however, the initial structure was gradually recovered over time after a sudden flooding.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.1
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• pp.66-74
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• 2012

The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and ${\beta}$-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 ${\mu}g$/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 ${\mu}g$/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 ${\mu}g$/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of $^{15}N$ into particle-associated microbial N ($^{15}N$-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) $^{15}N$-PAMN at 4 h only, but EFE on AS increased (p<0.001) $^{15}N$-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) $^{15}N$-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.7
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• pp.1021-1028
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• 2012

Peripheral blood mononuclear cells (PBMCs) discriminate microbial pathogens and induce T-cell responses of appropriate effector phenotype accordingly. Toll-like receptors (TLRs), in part, mediate this microbial recognition and differentiation while the development of T-cell effector functions critically depends on the release of Th1- or Th2- type cytokines. In the present study, buffalo PBMCs were stimulated under in vitro culture conditions by Bacillus subtilis cell wall petidoglycan, a TLR2 ligand, in a dose- and time- dependent manner. The expression of TLR2 as well as the subsequent differential induction of the Th1 and Th2 type cytokines was measured. Stimulation was analyzed across five doses of peptidoglycan ($10{\mu}g/ml$, $20{\mu}g/ml$, $30{\mu}g/ml$, $40{\mu}g/ml$ and $50{\mu}g/ml$) for 3 h, 12 h, 24 h and 36 h incubation periods. We observed the induction of TLR2 expression in a dose- and time-dependent manner and the peptidoglycan induced tolerance beyond $30{\mu}g/ml$ dose at all incubation periods. The correlation between peptidoglycan stimulation and TLR2 induction was found positive at all doses and for all incubation periods. Increased production of all the cytokines was observed at low doses for 3 h incubation, but the expression of IL-4 was relatively higher than IL-12 at the higher antigen doses, indicating tailoring towards Th2 response. At 12 h incubation, there was a pronounced decrease in IL-4 and IL-10 expression relative to IL-12 in a dose- dependent manner, indicating skewing to Th1 polarization. The expression of IL-12 was highest for all doses across all the incubation intervals at 24 h incubation, indicating Th1 polarization. The relative expression of TNF-${\alpha}$ and IFN-${\gamma}$ was also higher while that of IL-4 and IL-10 showed a decrease. For 36 h incubation, at low doses, relative increase in the expression of IL-4 and IL-10 was observed which decreased at higher doses, as did the expression of all other cytokines. The exhaustion of cytokine production at 36 h indicated that PBMCs became refractory to further stimulation. It can be concluded from this study that the cytokine response to sPGN initially was of Th2 type which skews, more pronouncedly, to Th1 type with time till the cells become refractory to further stimulation.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.185-199
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• 2017

Methane, which is emitted from natural and anthropogenic sources, is a representative greenhouse gas for global warming. Methanotrophs are widespread in the environment and play an important role in the biological oxidation of methane via methane monooxygenases (MMOs), key enzymes for methane oxidation with broad substrate specificity. Methanotrophs have attracted attention as multifunctional bacteria with promising applications in biological methane mitigation technology and environmental bioremediation. In this review, we have summarized current knowledge regarding the biodiversity of methanotrophs, catalytic properties of MMOs, and high-cell density cultivation technology. In addition, we have reviewed the recent advances in biological methane mitigation technologies using methanotrophs in field-scale systems as well as in lab-scale bioreactors. We have also surveyed information on the dynamics of the methanotrophic community in biological systems and discussed the various challenges pertaining to methanotroph-related biotechnological innovation, such as identification of suitable methanotrophic strains with better and/or novel metabolic activity, development of high-cell density mass cultivation technology, and the microbial consortium (methanotrophs and non-methanotrophs consortium) design and control technology.

• Korean Journal of Organic Agriculture
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• v.8 no.3
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• pp.131-146
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• 2000

This experiment was focused on finding out the potential problems in organic farming system, which is recently emphasized on the produce of horticultural crops due to the serious environmental deterioration. Thus, in this study, the effect of the application of organic farming materials in soil on plant growth and components of chinese cabbage and the soil Physicochemistry property were compared with conventional culture. The results were summarized as follows : 1. The effect of soil chemical properties after application of organic farming materials, The amounts of pH and O.,M. in N, P, K treated plot were few of change, To the contray, fermentation compost and microorganisms fermentation compost treated plot were some increase, Amount of P$_2$O$_{5}$ , Ca and increase in comparison with the N, P, K treated plot, Specially chicken manure+ microorganisms fermentation compost treated plot was the highest. 2. Change of soil microbial after application of organic farming materials, The number of bacteria and actinomycetes in N, P, K treated plot were appeared definite direction, to the fermentation compost and microorganisms fermentation compost treated plot showed the increased tendency, specially, chicken manure+ microorganisms fermentation compost treated plot was the highest, 3. The growth characteristic of chinese cabbage after application of organic farming materials was superior in order of chemical fertilizers plot> microorganisms fermentation compost plot> fermentation compost plot. It shows the NO$_3$$^{[-10]}$ accumulation in chinese cabbage was higher in the outer leaves than in the inner leavess. It was some lower in the microorganisms fermentation compost plot, it was the highest at chemical fertilizers plot , all inner and outer leaves.

• Korean Journal of Environmental Agriculture
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• v.29 no.3
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• pp.287-292
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• 2010

We aimed to investigate the responses of soil organisms to cover plants and to provide information for the selection of proper plant species. We studied the effects of 7 cover plants, including rye, oat, rattail fescue, Chinese milk vetch, red clover, crimson clover, and hairy vetch, on soil organisms in an apple orchard. An increase in the microbial phospholipid fatty acids (PLFA) and in the number of nematodes and microarthropods in the soil under the cover plants reflects elevated activities of soil organisms. A decrease in the level of some marker PLFA, which is an indicator of environmental stress, suggests that cover plants provide favorable environments for soil organisms. The population of fungi and animals that feed on fungi increased in the soil surface under red clover. The population density of nematodes and mites increased in the soil surface under rattail fescue, and that of mites and omnivorous nematodes increased in the soil surface under Chinese milk vetch. The level of microbial PLFA in the soil surfaces under the tested cover plants was higher than that under clean culture system. These results suggest that proper selection of the cover plants can facilitate the creation of favorable environments for soil organisms.

• KSBB Journal
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• v.21 no.4
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• pp.306-311
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• 2006

Cell preservation is indispensable in animal cell culture process and should be established according to the cell characteristics. In this study, we experimented hypothermic preservation of CHO cells that is widely used in pharmaceutical industry to produce therapeutic proteins and established a stable method of preservation. The highest viability of CHO cells was obtained when the cells were preserved using rolling tube, which means the cells should be suspended to avoid the cell lumping during the preservation. Also, we obtained superior preservation result under the anaerobic condition. To evaluate the serum-free media as a preservation solution, we investigated cell growth after hypothermic preservation using serum-free media. High cell viability and normal cell growth was observed during 10 days using serum-free media. Moreover, we found that more effective preservation when ${\alpha}$-tocopherol and retinoic acid is added to media as an additive. In the case of 1 liter large scale hypothermic preservation using established protocol, cell viability and growth rate was obtained as good as small scale one. This study is considered to be helpful for hypothermic preservation of CHO cells and large scale hypothermic preservation may be available through the further studies.

• Korean Journal of Food Science and Technology
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• v.48 no.6
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• pp.548-554
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• 2016

Since some strains of kimchi lactobacilli can curdle milk, they can be used for making yogurt. However, the best Lactobacillus plantarum strains for curdling milk for yogurt are still unknown. In this study, we determined the best L. plantarum strains for curdling milk, and the physicochemical properties of yogurts made using different L. plantarum strains were examined. Three strains of L. plantarum useful for curdling milk were identified (YD2, YD9, YD12). The number of lactobacilli was lower in yogurts made with L. plantarum than in those made with control, and among the L. plantarum strains tested, YD12 had the highest bacterial counts. However, the microbial count reached $6.3{\times}10^8CFU/mL$ after 24-h fermentation in all yogurts. The pH of the yogurts decreased after 12-h fermentation, while the acidity increased. The low pH and high acidity decreased the viscosity in all the three types of yogurts, because the acids disturbed gel formation due to protein denaturation. Sensory evaluation revealed that the YD12 group showed a high percentage of completion similar to the control group. YD2 and YD9 showed a high sourness value and low sweetness value, whereas YD12 yielded optimal values for all the organoleptic characteristics. Therefore, YD12 would be a high quality bacterial strain for use as a yogurt starter culture.

• Korean Journal of Microbiology
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• v.48 no.3
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• pp.192-199
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• 2012

Microalgal biotechnology has gained prominence because of the ability of microalgae to produce value-added products including biodiesel through photosynthesis. However, carbon and nutrient source is often a limiting factor for microalgal growth leading to higher input costs for sufficient biomass production. Use of municipal wastewater as a low cost alternative to grow microalgae as well as to treat the same has been demonstrated in this study using mini raceway open ponds. Municipal wastewater was collected after primary treatment and microalgae indigenous in the wastewater were encouraged to grow in open raceways under optimum conditions. The mean removal efficiencies of TN, TP, COD-$_{Mn}$, $NH_3$-N after 6 days of retention time was 80.18%, 63.56%, 76.34%, and 96.74% respectively. The 18S rRNA gene analysis of the community revealed the presence of Chlorella vulgaris and Scenedesmus obliquus as the dominant microalgae. In addition, 16S rRNA gene analysis demonstrated that Rhodobacter, Luteimonas, Porphyrobacter, Agrobacterium, and Thauera were present along with the microalgae. From these results, it is concluded that microalgae could be used to effectively treat municipal wastewater without aerobic treatment, which incurs additional energy costs. In addition, municipal wastewater shall also serve as an excellent carbon and nitrogen source for microalgal growth. Moreover, the microalgal biomass shall be utilized for commercial purposes.