• Title/Summary/Keyword: inoculation method

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Studies on the Tuberculin - 2) Serological observation of hemagglutinating antibody in guinea pigs sensitized with heat killed tubercle bacilli (Tuberculin에 관한 연구(硏究) - (2) 항원감작적혈구응집반응(抗原感作赤血球凝集反應)에 의한 결핵사균면역(結核死菌免疫) Guinea pig 혈제(血濟)의 적혈구응집(赤血球凝集) 항체가(抗體價) 소장(消長))

  • Kim, Jung-Kyu
    • The Journal of the Korean Society for Microbiology
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    • v.5 no.1
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    • pp.41-48
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    • 1970
  • In order to assay one lot of the potency of tuberculin for intradermic use, four guinea pigs sensitized with heat killed M. tuberculosis were used by the method originally of the Agriculture Research Service Center(A. R. S.) and Bureau of Animal. Industry(B. A. I.) in the United. States of America in which guinea pigs were used on 21st day after sensitization. There is doubt whether the guinea pigs are sensitized or not. Therefore, this study has taken the method of Middle Brook-Dubos hemagglutination test, and observed the humoral hemagglutinating antibody in the blood. The results obtained are as follows. 1. The maximum titer of hemagglutinating antibody was demonstrated 30days after sensitizing the guinea pigs with killed tubercle bacilli. The maximum titer was continued for a period of 45 days and thereafter declined gradually. 2. The swelling and redness of the region of the sensitized guinea pigs was shown to be reached to the maximum size within 24 hours after intradermal inoculation with HCSM tuberculin and gradually reduced 96 hours after inoculation. The swelling and redness could not be detected by macro-observation after 14 days.

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Effects of Microstructure Morphology on Fluid Flow Characteristics of A356 Commercial Alloy in Semi-Solid Slurry (반고상 A356 합금 슬러리의 미세조직에 따른 유동특성에 관한 연구)

  • Kim, Jae-Min;Lee, Seung-Hoon;Hong, C.P.
    • Journal of Korea Foundry Society
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    • v.25 no.6
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    • pp.240-248
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    • 2005
  • The rheocasting characteristics are strongly influenced by the microstructural morphology such as particle size, form factor and contiguity. In this study, the effect of structural morphology on fluid flow characteristics of A356 semi-solid alloy was investigated with a vacuum suction fluidity test. Semi-solid metal slurry was made by the mechanical stirring, the liquidus casting, and H-NCM to be analysed. H-NCM could obtain uniform and fine globular microstructures of 0.9 form factor and 55 ${\mu}m$ particle size. Inoculation was found to be effective for reducing particle size, however, for H-NCM it should be avoided due to the cause of increasing contiguity. The fluidity test indicated that the non-stirring method had higher fluidity and smaller liquid segregation in the same solid faction of 0.4 than the stirring method, for smaller particle size and higher form factor. It was observed that liquid segregation decreased as the particle size is smaller and form factor is higher. The results of die-casting experiment were a good agreement with those of fluidity test.

Production of Pleurotus ostreatus and Flammulina velutipes Using Liquid Spawn Inoculation System (느타리 팽나무버섯 재배에서 액체종균 배양 및 접종시스템 적용방법의 구명)

  • Hong, Sung-Jun;Lee, Won-Ho;Shin, Beom-Soo;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.31 no.1
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    • pp.22-27
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    • 2003
  • This research aimed at developing the efficient method and device applicable to the inoculation of mushroom using (spawn was intended to utilize) liquid spawn for the stable production of mushroom. For the mass production of liquid culture, optimal inoculum volume and cultural period were $5{\sim}6%$ and 4 days for Flammulina velutipes and 4% and 5 days for Pleurotus ostreatus. Fruit body weight in 850 ml polythene bottle was highest at 6% or 15 ml liquid inoculum for P. ostreatus and $4{\sim}6%$ or $10{\sim}15ml$ for F. velutipes. Weight of fruit body by the application of liquid spawn inoculation system increased up to 33.7% for P. ostreatus and 32.8% for F. velutipes, respetively, compared to conventional spawn making system. The system of liquid spawn inoculation was successfully operated without malfunction in opening or closing the lid, and it took 26 min to inoculate 1200 bottles.

Effect of the Mixed Culture of Bacillus subtilis and Lactobacillus plantarum on the Quality of Cheonggukjang (Bacillus subtilis와 Lactobacillus plantarum의 혼합배양이 청국장의 품질에 미치는 영향)

  • Ju, Kyung-Eun;Oh, Nam-Soon
    • Korean Journal of Food Science and Technology
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    • v.41 no.4
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    • pp.399-404
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    • 2009
  • The goal of this study was to improve the quality of cheonggukjang by the optimization of the inoculation methods of the Bacillus subtilis (B. subtilis) and Lactobacillus plantarum (L. plantarum) strains. In order to optimize the mixed cultivation of B. subtilis and L. plantarum, the B. subtilis strain was inoculated into steamed soybeans after cultivation of L. plantarum. Inoculation size of B. subtilis was changed to the simultaneous inoculation method in order to stimulate the growth of the L. plantarum in cheonggukjang. The viable cell count of L. plantarum increased from $2{\times}10^7$ CFU/g to $2-6{\times}10^8$ CFU/g and B. subtilis grew to $9{\times}10^8$ CFU/g. These results showed that 2 strains were successfully able to grow in the steamed soybean for good quality of cheonggukjang by optimization of the inoculation methods. The sensory evaluation indicated that a favorable aroma and overall acceptance of cheonggukjang by the optimized mixed cultivation of B. subtilis and L. plantarum, which was relatively higher than those of cheonggukjang by single strain inoculation of B. subtilis.

Development of Effective Screening Method and Evaluation of Radish Cultivars for Resistance to Fusarium oxysporum f. sp. raphani (효율적인 무 시들음병 저항성 검정법 개발 및 무 품종들의 병 저항성 평가)

  • Baik, Song-Yi;Kim, Jin-Cheol;Jang, Kyoung-Soo;Choi, Yong-Ho;Choi, Gyung-Ja
    • Research in Plant Disease
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    • v.16 no.2
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    • pp.148-152
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    • 2010
  • To establish the efficient screening method for resistance of radish to F. oxysporum f. sp. raphani, we investigated the development of Fusarium wilt of two radish cultivars, 'Songbaek' (susceptible) and 'Tokwang' (moderately resistant), according to several conditions such as inoculation methods, inoculum concentrations, and dipping periods of radish roots in spore suspension. By infected soil and soil-drenching inoculation methods, Fusarium wilt did not occur on the seedlings of both cultivars. In root dipping inoculation method using cut or non-cut roots of radish plants, the cut roots were easily infected by the pathogen than non-cut roots. And the disease development of two cultivars represented significant difference in non-cut root method. On the other hand, disease severity of Fusarium wilt on radish seedlings according to inoculum concentration increased in a dose-dependant manner, regardless of dipping periods. Using screening method established from the results, the 41 commercial radish cultivars were evaluated the degree of resistance to F. oxysporum f. sp. raphani. Among them, 6 radish cultivars were resistant, 22 cultivars were moderately resistant, and 13 cultivars were susceptible to Fusarium wilt.

Improving Pneumovirus Isolation Using a Centrifugation and AZD1480 Combined Method

  • Lee, Hansaem;Woo, Hye-Min;Kim, Kisoon;Park, Sehee;Park, Man-Seong;Kim, Sung Soon;Kim, You-Jin
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.2006-2013
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    • 2019
  • The isolation of respiratory viruses, especially from clinical specimens, often shows poor efficiency with classical cell culture methods. The lack of suitable methods to generate virus particles inhibits the development of diagnostic assays, treatments, and vaccines. We compared three inoculation methods, classical cell culture, the addition of a JAK2 inhibitor AZD1480, and centrifugation-enhanced inoculation (CEI), to replicate human respiratory syncytial virus (HRSV) and human metapneumovirus (HMPV). In addition, a combined method using AZD1480 treatment and CEI was used on throat swabs to verify that this method could increase virus isolation efficiency from human clinical specimens. Both CEI and AZD1480 treatment increased HRSV and HMPV genome replication. Also, the combined method using CEI and AZD1480 treatment enhanced virus proliferation synergistically. The combined method is particularly suited for the isolation of interferon-sensitive or slowly growing viruses from human clinical specimens.

Determination of an Effective Method to Evaluate Resistance of Bottle Gourd Plant to Fusarium oxysporum f. sp. lagenaria (박 덩굴쪼김병 저항성 검정조건 구명)

  • Kim, Sang Gyu;Lee, Oak Jin;Lee, Sun Yi;Kim, Dae Young;Huh, Yun-Chan;An, Se Woong;Jang, Yoon ah;Moon, Ji hye
    • Journal of Bio-Environment Control
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    • v.29 no.1
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    • pp.96-102
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    • 2020
  • Fusarium wilt caused by Fusarium oxysporum is a devastating disease limiting production of watermelon in Korea. The best way to control diseases is to use resistant gourd rootstock on watermelon. This study was conducted to establish an efficient screening method for resistant bottle gourd to Fusarium oxysporum f. sp. lagenaria. To develop an efficient inoculation method, incubation temperature after inoculation (15, 20, 25, and 30℃), inoculum concentration (1 × 105, 5 × 105, 1 × 106, and 5 × 106 conidia·mL-1), and growth stages of seedlings (7, 10, 13, and 16 days) was investigated. Disease development of Fusarium wilt of bottle gourd was little affected by differences in incubation temperature and growth stages of seedlings. But resistant lines were more susceptible and appeared more severe symptoms at the higher inoculation level. Taken together, we suggest that an effective screening method for resistant gourd plant to Fusarium wilt is to dip the roots of 10-day old seedlings in spore suspension of 1 × 105 - 1 × 106 conidia·mL-1, for 30 min, to transplant the seedlings into a non-infected soil, and then to incubate the inoculated plants in a growth room at 25℃ for 3 weeks to develop Fusarium wilt.

An Evaluation Method for Sesame (Sesamum indicum L.) Resistance to Phytophthora nicotianae var. parasitica (참깨발병 품종저항성 검정방법 연구)

  • Seong Ho, Choi;Eui Kyoo, Cho;Young Am, Chae
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.32 no.2
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    • pp.173-180
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    • 1987
  • Symptom development and disease severity of Phytophthora blight in the sesame plants varied depending upon age of the plants tested, inoculation method, watering method, and inoculum density in both susceptible Suweon 9 and Suweon 26 and moderately resistant B-67 and IS 103 sesame lines to Phytophthora nicotianae var. parasitica when inoculated. However, successful differentiation of the sesame lines for varietal resistance was possible using 20-day old seedling, inoculation by soil infestation, saturated soil water condition by half immersion of pots in water tank, and 200 sporangia per one ml of inoculum. Spraying or soil inoculation to 70-day old plants also was effective in differentiating the varietal resistance. By the screening method Suwon 26 showed 100% diseased plants and symptom severity index 9.0, while B-67 showed 20% diseased plants and symptom severity index 1.7. The rating scale given was from 0 through 9. For example, the scale 0 signified no symptom development, 5 signified discoloration of basal part of stem, and 9 signified discoloration of stem more than 10 cm high above the soil surface with blighted leaves. Differentiation in symptom severity also was made by percentage of the lesion area. Results evaluated using both parameters were well corresponded in varietal reaction of sesame to Phytophthora blight.

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Establishment of Pathogenicity Test Method for Macrophomina phaseolina Causing Soybean Charcoal Rot (콩균핵마름병균에 대한 병원성 검정법 확립)

  • So Hyeon An;Heung Tae Kim
    • Research in Plant Disease
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    • v.29 no.1
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    • pp.1-10
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    • 2023
  • The establishment of a laboratory assay and a greenhouse assay was conducted for evaluating the pathogenicity of Macrophomina phaseolina causing soybean charcoal rot established. In the laboratory assay, microsclerotia and hyphae were used as inoculum. In the laboratory assays using microsclerotia as an inoculum, disease incidences of M. phaseolina NSW17-108 and HSM17-034 were higher at 35℃ than 25℃. Of the two isolates NSW17-108 and HSM17-034, the disease incidence of HSM17-034 isolated from diseased sesame is higher than that of NSW17-108 isolated from diseased soybean. When the mycelia of M. phaseolina were used as an inoculum, the disease incidence of NSW17-108 and HSM17-034 at 35℃ exceeded 80% even after only 5 days of inoculation. Even at 25℃, furthermore, that of HSM17-034 exceeded 80% 5 days later. In the pathogenicity assays at a greenhouse, toothpicks where microsclerotia were produced or microsclerotia harvested from potato dextrose agar medium were used as an inoculum. In all greenhouse assays, M. phaseolina NSW17-108 and HSM17-034 showed 40-60% of disease incidences 35-65 days after inoculation with the pathogen, depending on the inoculation method. Between the two isolates, the pathogenicity of HSM17-034 was stronger than that of NSW17-108, and this result was consistent with laboratory assay results. Since the laboratory and greenhouse test methods tested in this study have different advantages and disadvantages depending on each test method, it is thought that the test method that can meet the purpose of the study should be selected and used.

Antimycotic Susceptibility Testing of Trichophyton Rubrum by Microculture Method (96-well microplate를 이용한 Trichophyton Rubrum의 항진균제 감수성검사)

  • Lee, Moo-Woong;Kim, Jong-Chul;Choi, Jong-Soo;Kim, Ki-Hong
    • Journal of Yeungnam Medical Science
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    • v.9 no.2
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    • pp.396-406
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    • 1992
  • Various susceptibility tests have been used to determine minimal inhibition concentration(MIC) of dermatophytes. They have limitations to apply practically because they need long time to determine MIC. Authors examined MIC of T. rubrum to ketoconazole and itraconazole using 96-well microplate and 24-well macroplate by method of Granade and Artis and tried to check the possibility of this method on clinical application. Nine strains of T. rubrum from patients with dermatophytosis were used. Evaluations of the factors affecting MIC were also tried. The results were as follows. 1. Effect of inoculation density on determination time and MIC : Determination of MIC were possible in 4th days after inoculation at higher inoculation density Caborbance 2.0, 1.0) compared to 6th days at lower inoculation density(absorbance 0.5, 0.25). 2. Effect of incubation temperature on MIC : When incubating at $37^{\circ}C$, MIC were below 0.006-$0.04{\mu}g/ml$ to ketokckonazole and below 0.006-$0.04{\mu}g/ml$ to itraconazole while at $25^{\circ}C$ 0.08-$5.68{\mu}g/ml$ to ketoconazole and 0.006-$0.71{\mu}g/ml$ to itraconazole. Significant reduction of MIC was observed at $37^{\circ}C$ compared to $25^{\circ}C$. 3. Effect of container size on determination time and MIC : When incubating in 96-well microplate and 24-well macroplate, determination of MIC was possible in 4th to 6th days after inoculation in broth-containig 96-well microplate compared to 8th to 12th days in broth-containing 24-well macroplate. But no difference in MIC was observed between different container size. 4. Effect of media on MIC : When using broth as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconazole, below 0.006-$0.36{\mu}g/ml$ to itraconazole in broth-containg 24-well macroplate. When using agar as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconzole, below 0.006-$5.68{\mu}g/ml$ to intraconzole in agar-containing 24-well macroplate. There was slight increase of MIC with agar media compared to broth media. 5. These findings confirm that determination of MIC of dermatophtes by method of Granade and Artis is fast and simple technique for antifungal susceptibility test.

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