• Korean journal of applied entomology
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• v.16 no.1 s.30
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• pp.47-53
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• 1977

Bacterial diseases of soybean has been recognized as a limiting factor of soybean production in Korea as it was estimated to cause around 10 percent of yield losses annually. The purpose of the study is to obtain information on the diseases through proving the kinds of pathogens and epidemiology, The wire brush method and multineedle appeared to be the best way of inoculation under all circumstances. Wire brush method, especially, was effective in shortening the incubation period and manifesting the lesion development by introducing more inoculum per unit of area. In case of spray inoculation it was necessary to apply a small amount(1 : 1,000) of wetting agent, twin-20, otherwise it was unabled to produce the diseases under field conditions. Two kinds of bacterial diseases caused by Pseudomonas glycinea and Xanthomonas phaseoli var. sljense were found from surveyed areas in Kore. Wild fire disease on soybean caused by Pseudomonas tabaci had not detectable during the experiment although there were several reports on the disease from other countries. when the pathogens were introduced into sterile soil, bacterial leaf blight pathogen could exsisted until 30 days while bacterial pustule pathogen survived only 4 days under the natural conditions of later June. Both bacteria, however, could produce the disease after more than 10 months period of storage in refrigerator when they were exsisted in infected plant tissues. In warehouse, non-temperature controlled, the bacteria lose their infectability within 6 months period from October to April even though they exsisted in infected tissues. Surface infested seeds with the pathogens could not produced the diseases on seedling stages of soybean plants when the seeds were planted in sterile soil after inoculation by dipping the seeds into bacterial suspensions, although germination was depressed by the pathogens when the seeds were planted on agar media.

• Korean journal of applied entomology
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• v.16 no.1 s.30
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• pp.1-6
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• 1977

The 'Kresek' disease in Korea caused by Xanthomonas oryzae was first found from eight counties in Jeon-nam and Kyungnam province in 1976. The study has been carried mainly on the grouping of pathogens and reproduction of the symptoms on Milyang #23, the variety had shown severe damages at the fields, by using the isolates from Hwa-sun and Kwang-san counties where the first epidemics took place. 1. The 'Kresek' disease was found mainly on Milyang #23, a new variety, at Hwa-sun, Mu-an, Kang-jin. Yung-am, Gog-sung counties in Jeon-nam province and Jin-yang county in Kyung-nam province. 2. The groups of 'Kresek' causing pathogens were the same of those producing bacterial leaf blight symptoms such as group I , II, IN and V of Xanthomonas oryzae. 3. Seventeen out of 21 isolates from Hwa-sun county where the first and severe damage found belonged to group IV, 2 to group II and 2 to group V All of 5 isolates from Kwang-san county belonged to the group IN. 4. The 'Kresek' type symptom could reproduced within 5 days after inoculation to seedlings by using root tut, spray and needle inoculation methods. j. The most of effective method for the inoculation was root cut, and then were spray and needle method. respectively. The higher concentration of inoculum produced the higher disease incidence.

• The Korean Journal of Pesticide Science
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• v.20 no.1
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• pp.35-40
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• 2016

Recently, bacterial angular leaf spot disease, caused by Xanthomonas fragariae, causes severe damage in strawberry production and its' export to other countries, since the pathogen has been classified as an A2 quarantine pathogen. Typical the Angular Leaf Spot (ALS) disease represent that water-soaked angular spots symptoms, bacteria ooze exudate under relatively high humidity condition and later the spot become reddish brown on the leaf surface. The pathogen disseminated by irrigation water, infected mother plant and farmer's hand. In this study, we reported that rubbing inoculation method showed more effective in the pathogen dissemination than infiltration with needles, regardless the strawberry cultivars. Additionally, Dichloroisocyanurate (NaDCC) treatment in commercial strawberry fields provided reliable efficiency to reduce the bacterial angular leaf spot disease incidency and severity.

• Journal of Acupuncture Research
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• v.18 no.1
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• pp.186-201
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• 2001

Purpose ; In order to study the effect of Houttuyniae herbal acupuncture on growth of melanoma B 16 and immunity in mice, the Control group with normal saline acupunchure after subcutaneous inoculation of B16BL6 cells, the Sample I with Houttuyniae herbal acupuncture manufactured by distilled water extraction method after subcutaneous inoculation of B16BL6 cells and the Sample II with Houttuyniae herbal acupuncture manufactured by alcohol extraction method after subcutaneous inoculation of B16BL6 cells were divided. Methods ; To evauluate the effect of Houttuynias herbal acupuncture on growth of melanoma B16 in mice, the weight of mouse melanoma, body weight, spleen index( $\sqrt(Weight of spleen/Body weight){\times}100$), lymphocytes numbers in mouse peripheral blood and spleen tissue, the percentage of CD4+ T cells, CD8+ and the ratio of CD4+/CD8+ in mouse peripheral blood and spleen tissue are measured. Results ; In study of the weight of mouse melanoma, Houttuyniae herbal acupuncture groups(Sample I, Sample II) showed statistically significant inhibitory effect. And in study of effect of reduction of change in body weight of mouse, Sample I showed statistically significant inhibitory effect, too. In study of reduction of spleen index, lymphocytes numbers in mouse spleen tissue, and the percentage of CD4+ and the ratio of CD4+/CD8+ T cells in mouse peripheral blood, Houttuyniae herbal acupuncture groups(Sample I, Sample II ) showed with statistically significant inhibitory effect. Also, Sample I showed inhibitory effect of reduction of the percentage of CD8+ T cells in mouse peripheral blood and spleen tissue and Sample II showed inhibitory effect of reduction of the percentage of CD4+ and the ratio of CD4+/CD8+ T cells in mouse spleen tissue with statistical significance. Conclusions ; The inhibitory effect of reduction of lymphocytes numbers in mouse peripheral blood and IL-2 productivity, Houttuyniae herbal acupuncture groups(Sample I, Sample II) didn't show significant effect.

• Korean Journal of Environmental Agriculture
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• v.33 no.3
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• pp.220-230
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• 2014

BACKGROUND: Excessive metals in the soil have become one of the most significant environmental problems. Phytoremediation has received considerable attention as a method for restoring the contaminated soils. The microbes having remarkable metal tolerance and plant growth-promoting abilities could also play a significant role in remediation of metal-contaminated soils, because bioaugmentation with such microbes could promote phytoextraction of metals. Therefore, the present study was focused on evaluating the phytoextraction of heavy metals (Co, Pb and Zn) in Helianthus annuus (sunflower) induced by bioaugmentation of a phosphate solubilizing bacterium. METHODS AND RESULTS: A phosphate solubilizing bacterium was isolated from metal-contaminated soils based on the greater halo size (>3 mm) with solid NBRIP agar medium containing 10 g glucose, 5 g $Ca_3(PO_4)_2$, 5 g $MgCl_2{\cdot}6H_2O$, 0.25 g $MgSO_4.7H_2O$, 0.2 g KCl, 0.1 g $(NH_4)_2SO_4$ in 1 L distilled water. Isolated bacterial strain was assessed for their resistance to heavy metals; $CoCl_2.6H_2O$, $2PbCO_3.Pb(OH)_2$, and $ZnCl_2$ at various concentrations ranging from $100-400{\mu}g/mL$ (Co, Pb and Zn) using the agar dilution method. A pot experiment was conducted with aqueous solutions of different heavy metals (Co, Pb and Zn) to assess the effect of bacterial strain on growth and metal uptake by Helianthus annuus (sunflower). The impact of bacterial inoculation on the mobility of metals in soil was investigated under laboratory conditions with 50 mL scaled polypropylene centrifuge tubes. The metal contents in the filtrate of plant extracts were determined using an atomic absorption spectrophotometer (Perkinelmer, Aanalyst 800, USA). CONCLUSION: Inoculation with Enterobacter ludwigii PSB 28 resulted in increased shoot and root biomass and enhanced accumulation of Co, Pb and Zn in Helianthus annuus plants. The strain was found to be capable of promoting metal translocation from the roots to the shoots of H. annuus. Therefore, Enterobacter ludwigii PSB 28 could be identified as an effective promoter of phytoextraction of Co, Pb and Zn from metal-contaminated soils.

• Applied Biological Chemistry
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• v.42 no.2
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• pp.111-115
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• 1999

This study was attempted to develop technique of masking the pungent odor of chonggugjang using two-stage fermentation method with mixed pure cultures. Cooked soybeans were fermented with Lactobacilli or Aspergillus oryzae at $38^{\circ}C$ for 36 hrs, and then re-fermented with Bacillus subtilis for 12 hrs. The volatile compounds of chonggugjang were obtained with a SDE(Simultaneous steam Distillation and solvent Extraction) system and the extracts were identified by GC and GC-MS. The experimental results revealed the presence of 35 volatile compounds in control chonggugjang(only Bacillus subtilis inoculation). Among them, the major volatile compounds were 2,5-dimethyl pyrazine, 2,3,5-trimethyl pyrazine and 2-methyl pyrazine. Twenty-nine kinds of volatile compounds were in chonggugjang prepared by two-stage fermentation method with Lactobacilli and Bacillus subtilis(II), and major volatile compounds were identified to be 2,3,5-trimethyl pyrazine and 2,5-dimethyl Pyrazine. In chonggugjang(Asp. oryzae and then Bacillus subtilis inoculation(III)), the contents of hexadecanoic acid and 2-methyl pyrazine were patricularly high and the main components of chonggugjang fermented with rice straw(IV) were 2,5-dimethyl pyrazine, 2,3,5-trimethyl pyrazine etc. In conclusion, the flavor compounds such as 2, 5-dimethyl pyrazine, 2,3,5-trimethyl pyrazine and 2-methyl pyrazine were increased by the inoculation of Lactobacilli or Asp. oryzae, where as unpleasant odor components-butyric acid and valeric acid were decreased. Compared with volatile compounds of chonggugjang made from different soybean cultivars, the flavor compounds (2-methyl pyrazine 2,5-dimethyl pyrazine 2,6-dimethyl pyrazine 2,3-dimethyl pyrazine 2-acetyl pyrazine 2,3,5-trimethyl pyrazine 2-ethyl-3,5-dimethyl pyrazine) of chonggugiang prepared with Sinpaldalkong were high.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.845-858
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• 1996

The purpose of this study was to establish early diagnostic methods for the detection of Aujeszky's disease viral antigens and nucleic acid in nasal cells, and buffy coats from experimentally infected living pigs by a combination of immunocytochemistry, in situ hybridization with digoxigenin(DIG)-labled probe and electron microscopy. Forty days old piglets were inoculated intranasally with $10^{7.0}TCID_{50}$ of Aujeszky's disease virus (ADV, NYJ-1-87 strain). The viral antigens and nucleic acid of ADV were detected in nasal cells, and buffy coat for 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopical method. The results were compared with conventional methods such as a porcine Aujeszky's disease serodiagnostic(PAD) kit, neutralization test(NT) and virus isolation. 1. The viral antigens, nucleic acids and capsids of ADV were detected in nasal cells, buffy coats from 3 days to 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopy, respectively. 2. When viral antigens were detected by the immunocytochemical technique, a diffuse brown deposit was observed in the nucleus and cytoplasm of nasal cells, buffy coats and PK-15 cells under a microscope. 3. DIG-labeled DNA probe was prepared by amplification of conserved sequence of recombinant ADV-gp50 clone with polymerase chain reacction. When ADV-DNA was detected by ISH with DIG-labeled probe, purplish blue pigmentation were observed in the nuclei and cytoplasms of ADV-infected cells under a microscope. Positive signals were observed in nasal cells and in the buffy coat and PK-15 cells at the first day after inoculation. 4. Where ADV-capsids were detected by transmission electron microscopical method, aggregation of capsids was observed in the nuclei and cytoplasms of nasal cells, buffy coats and PK-15 cells. The results suggested that these methods were considered as the highly sensitive and reliable tools for rapid and confirmative diagnosis of Aujeszky's disease in living pigs.

• Parasites, Hosts and Diseases
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• v.54 no.1
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• pp.15-20
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• 2016

Toxoplasmosis is an important zoonotic disease that can cause abortion in humans and animals. The aim of this study was isolation and subsequent genotyping of Toxoplasma gondii isolates in ovine aborted fetuses. During 2012-2013, 39 ovine aborted fetuses were collected from sheep flocks in Khorasan Razavi Province, Iran. The brain samples were screened for detection of the parasite DNA by nested PCR. The positive brain samples were bioassayed in Webster Swiss mice. The serum samples of mice were examined for T. gondii antibodies by IFAT at 6 weeks post inoculation, and T. gondii cysts were searched in brain tissue samples of seropositive mice. The positive samples were genotyped by using a PCR-RLFP method. Subsequently, GRA6 sequences of isolates were analyzed using a phylogenetic method. The results revealed that T. gondii DNA was detected in 54% (20/37, 95% CI 38.4-69.0%) brain samples of ovine aborted fetuses. In bioassay of mice, only 2 samples were virulent and the mice were killed at 30 days post inoculation, while the others were non-virulent to mice. The size of cysts ranged $7-22{\mu}m$. Complete genotyping data for GRA6 locus were observed in 5 of the 20 samples. PCR-RLFP results and phylogenetic analysis revealed that all of the isolated samples were closely related to type I. For the first time, we could genotype and report T. gondii isolates from ovine aborted fetuses in Khorasan Razavi Province, Iran. The results indicate that the T. gondii isolates are genetically related to type I, although most of them were non-virulent for mice.

• Korean journal of applied entomology
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• v.14 no.1 s.22
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• pp.1-5
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• 1975

The study has been carried out for the development of an inoculation method in screening resistant varieties and/or lines to bacterial leaf blight of rice with special consideration on plant ages and inoculum concentrations. A higher incidence of the disease was found on younger plants than on the older ones when the plants were inoculated with the same concentration of inoculum by clipping method under the same circumstances. Applications of extremly high concentration of inoculum resulted the death of younger seedlings from all varieties within short period after inoculation, and the inoculum that was lower than optimum concentration failed in distinguishing varietal characteristics in regard of resistance. The proper inoculum size for the screening of resistance among the varieties and/or lines depended on the age of plants. The optimum concentration of inoculum for the plant age of 14, 37, 48 and 58days was $10^6,\;10^7,\;10^7-10^{-9}\;and\; 10^9 cells/ml.,$ respectively.

• The Journal of the Korean Society for Microbiology
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• v.7 no.1
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• pp.21-27
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• 1972

A simple and effective procedure is described for semi-purification of viable Myco. leprae from biopsied lepromatous nodules by trypsinization and high-speed centrifugation. An unique characteristic of this method is a complete omission of conventional grinding or homogenization of minced lepromatous tissues prior to purification. Inoculation of trypsin-purified preparation of Myco. leprae into foot pads and ear lobes of Korean chipmunk(Tamias sibiricus asiaticus, Gmelin) resulted in apparent increases in total number of acid-fast bacilli per inoculated tissue 8 and 12 months after inoculation.