• Title/Summary/Keyword: Inoculation density

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Development of ELISA for detection of canine brucellosis (Canine brucellosis 검출을 위한 ELISA 진단법 확립)

  • Hur, Jin;Baek, Byeong-Kirl
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.159-166
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    • 2011
  • This study was carried out to develope enzyme-linked immunosorbent assay (ELISA) for detection of canine brucellosis in dogs experimentally inoculated with Brucella abortus 1119-3 and B. canis RM666. Groups A, B and C of dogs (each group consisting of three dogs) were orally inoculated with approximately $5{\times}10^9$ colony-forming units of B. abortus and B. canis, and with sterile pyrogen-free PBS, respectively. The animals were monitored at regular intervals upto the 12th week post inoculation (PI) by standard tube agglutination test (STAT), plate agglutination test (PAT), Rose Bengal test (RBT), 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and ELISA. The induced antibody titers in group A dogs were detected from the first week PI to the eighth week PI in STAT, PAT and RBT using the inactivated whole cells of B. abortus 1119-3 as antigens, while no sera in groups B and C dogs reacted with the antigens. In 2ME-RSAT using whole cells of B. canis M-strain as antigens, the induced antibody titers in group B dogs were observed at the second week PI and persisted for the 12th week PI, while sera of groups A and C dogs did not react with the whole cells. In ELISA using cytoplasmic fractions antigen of B. abortus 1119-3, the mean optical density of antibodies in groups A and B was detected from the first and second weeks PI, respectively, and persisted for 12th week PI, while sera of group C did not cross-react with the fractions antigen. However, in ELISA using the hot saline extracts of B. canis M- as an antigen, the induced antibody titers in only group B dogs were detected from second week PI and persisted for until the end of this study. These results indicate that the ELISA using B. abortus 1119-3 cytoplasmic fractions as antigens can be a good candidate for detection of brucellosis by B. abortus as well as B. canis in dogs.

Antimicrobial Activity of Hwangryunheadok-tang, and Scutellariae Radix Extract, Phellodendri Cortex Extract, Coptis Rhizoma Extract, Gardenia Jasminoides Extract against Staphylococcus aureus (황련해독탕(黃連解毒湯), 황련(黃連), 황금(黃芩), 황백(黃柏), 치자(梔子)의 Staphylococcus aureus에 대한 항균효과)

  • Kim, In-Soo;Oh, Hyun-Seung;Shin, Min-Koo;Lee, Young-Su
    • The Journal of Internal Korean Medicine
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    • v.35 no.4
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    • pp.455-471
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    • 2014
  • Objectives: This experiment was done for investigating antimicrobial activity of Hwangryunheadok-tang and Scutellariae Radix extract, Phellodendri Cortex extract, Coptis Rhizoma extract, Gardenia Jasminoides extract against Staphylococcus aureus. Methods: After administering S. aureus on a bacterial culture media plate, antimicrobial activity was tested by dripping $80{\mu}l$ diluted Hwangryunheadok-tang and Scutellariae Radix extract, Phellodendri Cortex extract, Coptis Rhizoma extract, and Gardenia Jasminoides extract (100%, 50%, 10%, 1%) on plates that were cultivated for a span of time from 16 to 72 hours. Also, minimal inhibitory concentration (MIC) was tested by dripping the minimum dilution density solution that has antimicrobial activity between $80{\mu}l$ and $20{\mu}l$ ($80{\mu}l$, $60{\mu}l$, $40{\mu}l$, $20{\mu}l$) in measure of density. Antimicrobial activity of Hwangryunheadok-tang and four kinds of extracts against S. aureus was continually displayed. Results: 1. S. aureus (Standard Microorganism, ATCC) (1) Antimicrobial activity was displayed for Hwangryunheadok-tang, Scutellariae Radix extract, and Phellodendri Cortex extract respectively in the undiluted solution and 50% of diluted magnification. Gardenia Jasminoides extract showed its activity only in the undiluted solution; Coptis Rhizoma extract showed its activity down to 10% of diluted magnification. The antimicrobial activity of the undiluted solution was increased when the volume of inoculation increased. But, there was no difference when time was extended for cultivation. (2) MIC of Hwangryunheadok-tang, Scutellariae Radix extract was 50%, $20{\mu}l$. Coptis Rhizoma extract was 10%, $20{\mu}l$, Phellodendri Cortex extract was 50%, $80{\mu}l$ and Gardenia Jasminoides extract was 100%, $60{\mu}l$. 2. S. aureus isolated from diarrheal patients (1) When compared to standard microorganism, MIC has decreased. As a result, their antimicrobial activity has increased. (2) Antimicrobial activity of Hwangryunheadok-tang and four extracts was continually shown in extending of the time, 16, 24 and 72 hours. Conclusions: The author comes to the conclusion that Hwangryunheadok-tang, and four kinds of extracts have antimicrobial activity against S. aureus. Especially, when comparing standard microorganisms with S. aureus isolated from diarrheal patients, some cases showed that antimicrobial activity of all the extracts were better than antibiotics. Thus, if a further study is performed, the use of Hwangryunheadok-tang, and four kinds of extracts will be valuable and beneficial in clinical treatments.

A Mechanism of Density-Dependent Population Change in Heterodera glycines (콩시스트선충의 밀도변화 기작)

  • Kim Young Ho;Riggs Robert D.;Kim Kyung Soo
    • Korean Journal Plant Pathology
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    • v.2 no.3
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    • pp.199-206
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    • 1986
  • Penetration level, female development and histological changes in infected root tissues were investigated following inoculation with different inoculum levels (110,440 and 1760 juveniles/plant) of Heterodera glycines (SCN) race 3 on susceptible 'Lee' and resistant 'Pickett' soybean cultivars. Penetration level was lower in Pickett at the higher inoculum levels but no differences were detected in Lee. However, the lower penetration level in the resistant soybean cultivar appeared not to be directly related to plant resistance (female maturation). The number of females recovered from Lee was lower at the highest inoculum level. The number of females maturing on Pickett was much less than that on Lee, showing that changes of SCN population is associated with the number of SCN maturing rather than nematode penetration. In Lee mono-infection sites (a single nematode per site) syncytia had dense cytoplasm and no central vacuoles. while multi-infected sites (many overlapping nematodes per site) had syncytia with a large central vacuole and many small vacuoles. Resistant responses in mono-and multi-infected root tissues of Pickett were delayed and rapid necrosis, respectively. The differences in tissue response are suggested as a mechanism that controls density-dependent population changes in resistant and/or susceptible soybean cultivars.

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Effect of Apple Leaf Hair Density on the Occurrence and Infection of Alternaria Leaf Spot Caused by Alternaria mali Roberts (모용(毛茸)이 사과나무 반점낙엽병(斑點落葉病) 발생(發生)과 병원균(病原菌)의 감염(感染)에 미치는 영향(影響))

  • Yoon, J.T.;Lee, J.T.
    • Korean journal of applied entomology
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    • v.26 no.4 s.73
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    • pp.245-249
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    • 1987
  • In the 82 apple varieties tested, the distribution of varieties grouped on the damage degree to Alternaria leaf spot were 15 varieties(l8.3%), which damaged above 50%, and 51 varieties(62.2%) damaged below 5%. Susceptible varieties to Alternaria leaf spot were Raritan, Indo, Winesap and Starkrimson, moderately resistant varieties Sekaiichi and Starking, and resistant Jonathan, Tsugaru, Molliies, Jonagold, Gala etc. The leaf hair density on the backside of leaf were rarer inresistant varieties than susceptible one. In inoculation test, penetration and infection of Alternaria mali were easier on the backside than surface of apple leaf at the same varity, and infection rate with Alternaria mali were higher at removal treatment than non-removal one of leaf hairs, so we can guess that leaf hair are related to resistance of variety to Alternaria mali.

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Studies on the Cultural Characteristics of Poria cocos (복령(Poria cocos)의 배양학적(培養學的) 특성(特性)에 관한 연구(硏究))

  • Hong, In-Pyo;Lee, Min-Wong
    • The Korean Journal of Mycology
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    • v.18 no.1
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    • pp.42-49
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    • 1990
  • The cultural characteristics and some factors such as nutrient sources and supplements effecting on mycelial growth and density were investigated to study the possibility of an artificial cultivation of P. cocos. The optimum pH for P. cocos was 4.0-4.5. The optimal growth temperature ranged from $25^{\circ}C$ to $29^{\circ}C$. Myceial growth of P. cocos was better in SPD than PD media. Adding the nurient sources such as dextrose, yeast and potato infusion to pine extract media practically stimulated the mycelial growth and density of P. cocos comparing to pine extract media alone. When P. cocos was cultured on sawdust media added 3 different supplements composed of corn meal, rice bran and wheat bran, corn meal was the best and its percentage was 30 (w/w) for mycelial growth. On culturing in sawdust media added by varying the mixture ratio of them, the media mixed corn meal and wheat bran (3:1, w/w) supported more vigours for mycelial growth. In inoculation test to pine stem, the fungal growth was good in under or inside pine bark and xylem, but the sclerotium was not observed in the stem. Mycelial growth was also observed in central part of pine stem by cross section.

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Gall formation on different age, habitat, and parasite position in Rhus javanica L. (붉나무의 수령, 서식지 및 기생위치에 따른 오배자 형성)

  • Jo, Hyeong-Chan;Han, Kwang-Soo;An, Eun-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.4
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    • pp.304-311
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    • 2000
  • This study was conducted to investigate the ratio and number of the Rhus javanica gall formed by chinese sumac aphid, Scheleechtendalia chinensis, in several different conditions in which R. javanica lives. The density of 11 to 15-year-old R. javanica was more higher and the number of galls was also higher than 1 to 5- or over 16-year-old one at these sites. The formed gall according to four directions such as the East, West, South and North was founded the highest at the East and followed South, North, and West order. The formative rate of R. javanica gall did not differ in the mixture forest, and in the habitat having a heap of a little stone and mosses near a valley was better than in the site of road perimeter. Through an index of the gall diameter and the number of chinese sumac aphid, we evaluated the gall processing and development formed by the aphid. As the density of aphids increase, the size of the gall was also more plumped. The part of phyllotaxy of R. javanica gall in leaf and pinnate of was increased in the middle and upper part of terminal leaflet and leaflet was more occupied than wing. R. javanica which did not form a gall in natural environmental, formed gall by a artificial inoculation of S. chinensis. Some gall disappeared during a gall development from various environmental conditions.

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Biological Control of Tomato and Red Pepper Powdery Mildew using Paenibacillus polymyxa CW (Paenibacillus polymyxa CW를 이용한 고추 및 토마토 흰가루병 방제)

  • Kim, Yong-Ki;Choi, Eun-Jung;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Jee, Hyeong-Jin;Park, Jong-Ho;Han, Eun-Jung;Jang, Bo-Kyung;Yun, Jong-Cheul
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.379-387
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    • 2013
  • In order to improve practical utility of agro-microorganisms (AMs) which had been cultured and disseminated to promote plant growth and to control crop diseases, 51 isolates of AMs were collected from 18 agricultural extension centers in local government and screened for multi-functions such as antifungal activity, activities of phosphorus solubilization, IAA and siderophore production, nitrogen fixation, and hydrolytic enzyme activity. Finally we selected one isolate showing good antifungal activity and multi-functions related to plant growth and disease control. The selected isolate, Paenibacillus polymyxa CW, showed good inhibitory effect against plant pathogens, Pyricularia gresea, Colletotrichum acutatum, Fusarium oxysporum, Phomopsis sp., Aspergillus niger, Rhizoctonia solani and Phytophthora capsici. Suppressive effect of P. polymyxa CW against the used plant pathogens except for R. solani was much higher than that of P. polymyxa AC-1 storing in National Academy of Agricultural Science. We found P. polymyxa CW isolate showed good activity in siderophore and IAA formation, and nitrogen fixation. With P. polymyxa CW isolate, siderophore formation activity was similar to that of P. polymyxa AC-1, but IAA formation and nitrogen fixation activity was much higher than that of P. polymyxa AC-1. However neither P. polymyxa CW nor P. polymyxa AC-1 showed hydrolytic enzyme (chitinase, pectinase and cellulase) activity. The treatment of P. polymyxa CW with culture suspension of different cell density ($10^8$, $10^7$. $10^6$ cfu/ml) showed that the highest density reduced incidence of red pepper powdery mildew by 68.3% after 10 days of application. As application density of P. polymyxa CW was decreased, its control efficacy was proportionally decreased. In addition, when P. polymyxa CW was treated to control tomato powdery mildew at the same concentrations and their control effects were investigated after 7 days of inoculation, disease incidence was 0.03, 19.5, 45.7%, respectively, compared to 56.3% that of untreated check. Like red pepper powdery mildew, increase of application density of P. polymyxa CW resulted in increase of its control efficacy proportionally. P. polymyxa CW showed a density-dependent control efficacy against red pepper and tomato powdery mildews. Therefore we think that mode of action of the antagonist for suppressing two powdery mildew diseases might be antibiosis and density of more than $10^8cfu/ml$ was needed to control effectively the two diseases. On this basis, we think that P. polymyxa CW can be a promising control agent for suppressing powdery mildews of red pepper and tomato.

Optimal conditions and effects of prebiotics for growth and antimicrobial substances production of Lactobacillus brevis BK11 (Lactobacillus brevis BK11의 증식과 항균물질 생산을 위한 최적 배양조건 및 prebiotics의 영향)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.288-299
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    • 2015
  • Lactobacillus brevis BK11 obtained from Baikkimchi was selected to study the effects of culture medium, initial pH, atmosphere composition, incubation temperature and time, and prebiotics on growth and production of antimicrobial substances. Growth and antimicrobial substances production of L. brevis BK11 were significantly higher in MRS broth than in BHI or M17 broth. The production of cell mass, lactic acid, and bacteriocin by BK11 strain was at maximum in MRS broth adjusted to pH 6.0. Aerobic and microaerobic conditions were favored cell growth and antimicrobial substances production than anaerobic condition. Biomass and lactic acid production and antimicrobial substances activity of BK 11 were significantly better at 30 and $37^{\circ}C$ than at $25^{\circ}C$. Growth of the strain BK11 entered the stationary growth stage at 24 h after inoculation, and decreased after 36 h. Antimicrobial activities of cell-free culture supernatant and bacteriocin solution were highest when cultured in MRS broth with an initial pH 6.0 for 24-30 h at $37^{\circ}C$. In addition, the highest cell number and lactic acid and bacteriocin production were recorded in the presence of 1 and 2% (w/v) fructooligosaccharide (FOS), however, inulin and raffinose did not affect biological and physicochemical characteristics and antimicrobial activities of L. brevis BK11 cultures. According to these results, production of antimicrobial substances by L. brevis KB11 was closely associated with cell density. Under optimal conditions for antimicrobial substances production, L. brevis BK11 effectively inhibited the growth of Helicobacter pylori ATCC 43504.

Breeding of Resistant Cabbage 'CT-171' to Fusarium Wilt (시들음병 저항성 양배추 품종 'CT-171' 육성)

  • Song, Jun-Ho;Kim, Gi-Jun;Kim, Kyoung-Cheol;Han, Tae-Ho
    • Horticultural Science & Technology
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    • v.32 no.1
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    • pp.129-132
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    • 2014
  • A new resistant cabbage variety 'CT-171' to Fusarium wilt was bred by crossing $A337MSBC_5$ with resistance to the disease and 397-$G_6$ with good density and color. 'CT-171' was selected after combining ability, seed gathering and regional adaptability test in 2008. For breeding of resistant varieties, we investigated the development of Fusarium wilt on cabbage seedlings inoculated with Fusariumoxysporum f. sp. conglutinans by root dipping inoculation method. As a result, 'CT-171' showed higher resistance to Fusarium wilt than 'Asiaball' used as control. The maturity of new variety was 58 days and was faster than control and well suited for autumn cultivation because of cold resistance. The anthocyanin pigment of plant was not revealed. The weight, height and width of head were 1.5 kg, 14 cm and 15 cm, respectively and the core size was 5.7 cm and stable in various cultivation environments. 'CT-171' which showed good agricultural character and resistance to Fusarium wilt filed for variety protect right in Korea Seed & Variety Service on February 2013. The new variety will be appropriate for export and domestic consumption.

Studies on the fluorescent Pseudomoηas isolated from the wheat rhizosphere (소맥근권(小麥根圈)에서 분리(分離)한 형광성(螢光性) Pseudomonas spp. 에 관(關)한 연구(硏究))

  • Lee, Myung-Chol;Kim, Yong-Woong;Kim, Kwang-Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.23 no.2
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    • pp.152-159
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    • 1990
  • Four active strains, producing siderophore and antagonizing to soil plant disease fungi, were isolated and identified from the wheat rhizosphere to reduce the injurious effects of continuous cropping and study on the biological control. The obtained results were summerized as follow. 1. Four strains of fluorescent Pseudomonas were isolated from the wheat rhizosphere and identified as Pseudomonas fluorescens Biotype A(Ps-1,2,5) and Biotype B(Ps-3) That strains inhibitied growth of R. solani, F.oxysperum and F. solani in vitro 2. Optical density of pigment was maximum at 410nm. 3. Siderophore production by identified strains was decreased with addition of $Fe^{+3}$, although not decreased with addition of $Fe^{+2}$ 4. Pigment of Ps-1, 2 and 3 strain inhibitied growth of R. slani, F. oxysperum and F. solani but pigment of Ps-5 strain did not inhibitie growth of R. solani 5. Effect of inoculation was in order of Ps-2, 1, 5 and 3 strain through the dark culture method, and effect of Ps-1 and Ps-2 was greater than that of treatment of captan 50ppm.

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