• Title/Summary/Keyword: Inoculation methods

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Effect of Inoculation of Azospirillum brasilense and Methylobacterium oryzae on the Growth of Red Pepper Plant (고추의 생장에 미치는 Azospirillum brasilense 및 Methylobacterium oryzae 접종 효과)

  • Kim, Byoung-Ho;Sa, Tong-Min;Chung, Jong-Bae
    • Korean Journal of Environmental Agriculture
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    • v.30 no.2
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    • pp.223-228
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    • 2011
  • BACKGROUND: Rhizosphere bacteria may improve plant growth and productivity both by supply nutrients and hormonal stimulation. Although many experiments have shown improvements in plant growth with inoculation of bacterial cultures to the rhizosphere, the main obstacle in the applications of plant growth promoting rhizobacteria in a large scale is the inconsistency of the results. We tested the growth promoting effects of Azospirillum and Methylobacterium strains on red pepper plant. METHODS AND RESULTS: Red pepper seedlings were grown for 25 days in a growth media inoculated with A. brasilense CW903 or M. oryzae CBMB20. The seedlings were transplanted and grown for 45 days in pots with soil in a greenhouse, at half the recommended level of fertilizer. Bacterial culture, $4.0{\times}10^9$ for A. brasilense CW903 and $5.8{\times}10^8$ CFU for M. oryzae CBMB20, was applied in root zone soil periodically every 10 days during the experiment. Inoculation of M. oryzae CBMB20 significantly increased the red pepper plant growth in terms of leaf number, height and mass of shoot, or root mass compared to uninoculated control plants. Although beneficial effects of A. brasilense on plant growth of many crops were observed, the growthpromoting effect of A. brasilense CW903 on red pepper plant was not found in this study. CONCLUSION(s): The factors responsible for the irregularities in plant growth promoting of rhizobacteria are difficult to elucidate. Extensive inoculation experiments in the greenhouse and in the field should enable us to define the factors critical to obtain successful application of plant growth promoting rhizobacteria.

Circulating Tumor Cell Detection in Lung Cancer Animal Model

  • Chong, Yooyoung;Jung, Yong Chae;Hwang, Euidoo;Cho, Hyun Jin;Kang, Min-Woong;Na, Myung Hoon
    • Journal of Chest Surgery
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    • v.54 no.6
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    • pp.460-465
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    • 2021
  • Background: Metastasis and recurrence of primary cancer are the main causes of cancer mortality. Disseminated tumor cells refer to cancer cells that cause metastasis from primary cancer to other organs. Several recent studies have suggested that circulating tumor cells (CTCs) are associated with the clinical stage, cancer recurrence, cancer metastasis, and prognosis. There are several methods of isolating CTCs from whole blood; in particular, using a membrane filtration system is advantageous due to its cost-effectiveness and availability in clinical settings. In this study, an animal model of lung cancer was established in nude mice using the human large cell lung cancer cell line H460. Methods: Six-week-old nude mice were used. The H460 lung cancer cell line was injected subcutaneously into the nude mice. Blood samples were obtained from the orbital area before cell line injection, 2 weeks after injection, and 2 weeks after tumor excision. Blood samples were filtered using a polycarbonate 12-well Transwell membrane (Corning Inc., Corning, NY, USA). An indirect immunofluorescence assay was performed with the epithelial cell adhesion molecule antibody. The number of stained cells was counted using fluorescence microscopy. Results: The average size of the tumor masses was 35.83 mm. The stained cells were counted before inoculation, 2 weeks after inoculation, and 2 weeks after tumor excision. Cancer cells generally increased after inoculation and decreased after tumor resection. Conclusion: The CTC detection method using the commercial polycarbonate 12-well Transwell (Corning Inc.) membrane is advantageous in terms of cost-effectiveness and convenience.

Development of Effective Screening Method for Efficacy Test of Trunk Injection Agents Against Pine Wood Nematode, Bersaphelenchus xylophilus in Japanese Black Pine, Pinus thunbergii (곰솔(Pinus thunbergii)에서 소나무재선충(Bersaphelenchus xylophilus) 나무주사 약제의 효율적인 평가법 개발)

  • Shin, Won Sik;Jung, Young Hack;Lee, Sang Myeong;Lee, Chae Min;Lee, Chang Joon;Kim, Dong Soo;Mun, Il Sung;Lee, Dong Woon
    • The Korean Journal of Pesticide Science
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    • v.19 no.4
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    • pp.440-449
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    • 2015
  • Pine wood nematode (PWN), Bursaphelenchus xylophilus is one of the most serious pests of pine tree. Trunk injection of some nematicides in tree is well known as an effective control method. However there are some limitating factors which hindering the efficacy of trunk injection in field or potted tree. In this study we suggested easy and useful alternative screening methods of nematicides against PWN. Reproduction of PWN was influenced by tree twig moisture (high reproduction in high moisture trig) and paraffin coating was 78.6% reduced moisture loss in tested twig. There were no reproduction different in up and down site from infection site of twig at 1 month after inoculation of PWN and also distance (5 and 10 cm) from inoculation site of PWN did not influence the reproduction of PWN. Numbers of reproduced PWN were higher with decreasing diameter of twig. Numbers of reproduced PWN were similar to P. thunbergii and P. densiflora. However reproduction was increased depending on high inoculation density and longer propagation period. When inoculation of PWN on cut twig injected with emamectin benzoate 2.15% EC and morantel tartrate 8% SL in trunk of Pinus thunbergii in the field, PWN number were significantly reduced than untreated control. We suggest this screening method for PWN control agents.

Development of Efficient Screening Method for Resistance of Cabbage Cultivars to Black Rot Disease Caused by Xanthomonas campestris pv. campestris (양배추 검은썩음병에 대한 효율적인 저항성 검정법)

  • Lee, Ji Hyun;Kim, Jin-Cheol;Jang, Kyoung Soo;Choi, Yong Ho;Ahn, Kyoung Gu;Choi, Gyung Ja
    • Research in Plant Disease
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    • v.19 no.2
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    • pp.95-101
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    • 2013
  • Black rot caused by Xanthomonas campestris pv. campestris (Xcc) is one of the most serious diseases of crucifers world-wide. To establish the efficient screening method for resistant cabbage to Xcc, different inoculation methods, inoculation positions, growth stages of seedlings, and incubation temperatures after inoculation were investigated with the seven cabbage cultivars showing different resistance degrees to the pathogen. Clipping with mouse-tooth forceps was better inoculation method than piercing with 18 pins or cutting with scissors to distinguish the level of resistance and susceptibility. In inoculation using mouth-tooth forceps, clipping the edges of the leaves near veins is more effective than injuring the veins of the leaves directly. In addition, the inoculated plants kept at $22^{\circ}C$ showed more clear resistant and susceptible responses than those kept at 26 or $30^{\circ}C$. On the basis of the results, we suggest that an efficient screening method for resistance of cabbage cultivars to black rot is to clip the edges of the leaves near veins of the four-week-old seedlings with mouth-tooth forceps dipped in a suspension of Xcc at a concentration of $7{\times}10^7$ cfu/ml and incubate the inoculated plants in a growth room at $22^{\circ}C$ with 12-hr light a day.

Studies on the Immunization Against Newcastle Disease III. Investions on the Immunity of Newcatle Disease with Special Reference to Vaccination Program and Route (뉴캣슬병 면역에 대한 검토 III. 예방접종 프로그램과 접종경노를 중심으로 한 뉴캣슬병 면역능 검토)

  • 이학철;정유열
    • Korean Journal of Poultry Science
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    • v.8 no.2
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    • pp.77-89
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    • 1981
  • The experimental study was undertaken to confirm the effect of vaccination of birds with Newcastle disease (ND) vaccines on the Market by use of th. various vaccination programs. Sixteen groups of birds varying from 2 to f days of age, which were originated from hyper-immunised hens against ND were immunised by three different ways, a live vaccine only, a killed vaccine only, and the combination of a live and killed vaccine according to the each schedule of employed programs. In the administration of a live vaccine only, birds were immunized by one of following methods, the combination of intranasal and intraocular inoculation, intramuscular inoculation, via drinking water and the double inoculation by spray and drinking water application. Except for the double application, all the birds were vaccinated 2,3 or 4 times with two volumes of the virus dose (drinking water application) instructed by the commercial vaccine laboratory, until 21, 28 or 30 days of age, and all the immunized birds 19, 21 or 28 days postvaccination were challenged intramuscularly with 1.0$m\ell$ of 10,000 MLD per $m\ell$ of a virulent ND virus. In the administration of the combination of a live and killed vaccine, birds were immunized 2 or 3 times intranasally at first until 14 or 28 days of age with the same dose of the above experiment of a live vaccine, and then inoculated intramuscularly 1 or 2 times until 60 days of age with 1.0 $m\ell$ of a killed vaccine. And all immunized birds 11 days postvaccination were challenged with the same procedure of the above experiment. In the administration of a killed vaccine only, birds were immunized 3 times intramuscularly until 28 days of age with varied dose (0.2-0.5 $m\ell$) of a killed vaccine and all immunized birds 33 days postvaccination were challenged with the same procedure of the above experiment. The results obtained are summerised as follows: All birds vaccinated by using the combination of a live and killed vaccine program or a killed vaccin only appeared to be refractory. without any sign of illness, to the challenge exposure with 1.0$m\ell$ of 10,000 MLD per $m\ell$ of a virulent ND virus. On the other hand, the survival rates of birds of live vaccine groups immunized by a number of vaccine program such as Salsbury's day old program, 3-3-3 program, the Institute of Veterinary Reserch program and Multiple inoculation program, were 39.58%, 43.7%, 43.75% and 47.80%, respectively. And the survival rates of birds vaccinated with a live vaccine by 4 different ways of administration, i.e., double inoculation by water and aerosol application, intramuscular injection, intranasal instillation and via 4.inking water were 87.50%, 64.06%, 42.18% and 25.00%, respectively.

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Effects of lactic acid bacteria inoculation in pre-harvesting period on fermentation and feed quality properties of alfalfa silage

  • Ertekin, Ibrahim;Kizilsimsek, Mustafa
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.2
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    • pp.245-253
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    • 2020
  • Objective: To develop the fermentation quality and chemical composition of alfalfa (Medicago sativa Lam.) silage, plants were inoculated with different lactic acid bacteria (LAB) strains at field 24 hours before harvest. Methods: The treatment groups were as follow: silage without additive as a control and inoculated with each strains of Lactobacillus brevis (LS-55-2-2), Leuconostoc citerum (L. citerum; L-70-6-1), Lactobacillus bifermentans (L. bifermentans; LS-65-2-1), Lactobacillus plantarum (L. plantarum; LS-3-3) and L. plantarum (LS-72-2). All the silages were stored at 25℃. Parameters such as pH, microorganism and volatile fatty acid contents, crude protein, neutral detergent fiber, acid detergent fiber, net gas, metabolizable energy, organic matter digestibility, dry matter intake and relative feed value were measured to determine fermentation quality, chemical compositions and relative feed value of alfalfa silages. Results: Significant differences were found among the control and treated groups in terms of pH and microorganism contents at all opening times and crude protein, net gas, metabolizable energy and organic matter digestibility of final silage. The pH values ranged from 4.70 to 5.52 for all treatments and control silage had the highest value of overall treatments at T75d silages. Volatile fatty acid of silages was not influenced significantly by inoculations. However, lactic acid content of L. bifermentans (LS-65-2-1) was higher than the other treatments. The highest metabolizable energy and organic matter digestibility were recorded from L. citerum (L-70-6-1) inoculation. In addition, no significant differences were found among treatments in terms of neutral detergent fiber, acid detergent fiber, dry matter intake and relative feed value. Conclusion: Among the treated LAB isolates, L. bifermentans came into prominence especially in terms of organic acid composition and quality characters of silages.

The effect of Glomus intraradices on the physiological properties of Panax ginseng and on rhizospheric microbial diversity

  • Tian, Lei;Shi, Shaohua;Ma, Lina;Zhou, Xue;Luo, Shasha;Zhang, Jianfeng;Lu, Baohui;Tian, Chunjie
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.77-85
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    • 2019
  • Background: Glomus intraradices is a species of arbuscular mycorrhizal fungi that, as an obligate endomycorrhiza, can form mutually beneficial associations with plants. Panax ginseng is a popular traditional Chinese medicine; however, problems associated with ginseng planting, such as pesticide residues, reduce the ginseng quality. Methods: In this experiment, we studied the effect of inoculating G. intraradices on several physiological properties and microbial communities of ginseng. UV-Visible Spectrum method was used to detect physical properties. Denaturing gradient gel electrophoresis method was used to analyze microbial communities. Results: The results indicated that inoculation with G. intraradices can improve the colonization rate of lateral ginseng roots, increase the levels of monomeric and total ginsenosides, and improve root activity as well as polyphenol oxidase and catalase activities. We also studied the bacterial and fungal communities in ginseng rhizospheric soil. In our study, G. intraradices inoculation improved the abundance and Shannon diversity of bacteria, whereas fungi showed a reciprocal effect. Furthermore, we found that G. intraradices inoculation might increase some beneficial bacterial species and decreased pathogenic fungi in rhizospheric soil of ginseng. Conclusion: Our results showed that G. intraradices can benefit ginseng planting which may have some instructive and practical significance for planting ginseng in farmland.

Improved immune responses and safety of foot-and-mouth disease vaccine containing immunostimulating components in pigs

  • Choi, Joo-Hyung;You, Su-Hwa;Ko, Mi-Kyeong;Jo, Hye Eun;Shin, Sung Ho;Jo, Hyundong;Lee, Min Ja;Kim, Su-Mi;Kim, Byounghan;Lee, Jong-Soo;Park, Jong-Hyeon
    • Journal of Veterinary Science
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    • v.21 no.5
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    • pp.74.1-74.13
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    • 2020
  • Background: The quality of a vaccine depends strongly on the effects of the adjuvants applied simultaneously with the antigen in the vaccine. The adjuvants enhance the protective effect of the vaccine against a viral challenge. Conversely, oil-type adjuvants leave oil residue inside the bodies of the injected animals that can produce a local reaction in the muscle. The long-term immunogenicity of mice after vaccination was examined. ISA206 or ISA15 oil adjuvants maintained the best immunity, protective capability, and safety among the oil adjuvants in the experimental group. Objectives: This study screened the adjuvant composites aimed at enhancing foot-and-mouth disease (FMD) immunity. The C-type lectin or toll-like receptor (TLR) agonist showed the most improved protection rate. Methods: Experimental vaccines were fabricated by mixing various known oil adjuvants and composites that can act as immunogenic adjuvants (gel, saponin, and other components) and examined the enhancement effect on the vaccine. Results: The water in oil (W/O) and water in oil in water (W/O/W) adjuvants showed better immune effects than the oil in water (O/W) adjuvants, which have a small volume of oil component. The W/O type left the largest amount of oil residue, followed by W/O/W and O/W types. In the mouse model, intramuscular inoculation showed a better protection rate than subcutaneous inoculation. Moreover, the protective effect was particularly weak in the case of inoculation in fatty tissue. The initial immune reaction and persistence of long-term immunity were also confirmed in an immune reaction on pigs. Conclusions: The new experimental vaccine with immunostimulants produces improved immune responses and safety in pigs than general oil-adjuvanted vaccines.

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.