• Title/Summary/Keyword: inoculation studies

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Studies on Development of Antagonistic Microorganism by Cell Fusion - Biological control of disease - ) (세포융합에 의한 신 길항미생물 육종에 관한 연구 - 목초 병해의 생물학적 방제 -)

  • 최기춘;이영환;전우복
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.15 no.1
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    • pp.1-12
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    • 1995
  • This study was to investigate an effective biological control of forage diseases and provide a basic data and a model in improving variety of antagonistic bacteria, with growth promoting effect on forage, through cell fusion. The results obtained were summarized as follows; 1. The antagonistic himbacterium against soil-borne phathogenic fungi Fusarium oxysporum and Rhizoctonia solani was isolated from continuous cropping himsphere soil of forage, and its biological and physiological characteristics were investigated. This bacterium was identified as Bacillus subrilis and named BS 101. Another strain for cell fusion was Bacillus thur ingiensis ssp. kurstaki HD-I(BT 37669) with insecticidal crystal. 2. The auxotropic mutants of BS 101 and BT 37669 were derived after mutagenesis using N-methyl-N'nitro- Nitrosoguanidine(NTG) to give amino acid requirement marker. n e s e auxotropic mutants of BS 101 and BT 37669 were named BS 1013(his-) and BT 69(asp-), respectively. 3. The best protoplast requirement was obtained using DM 3 medium, containing 5% casamino acid, 1 M $MgCI_2$ and 2% bovine semm albumin, to give Fusant 3, 7 and 8. BT toxin gene was not identified with fusants by Southern blotting. However, SDS-PAGE analysis of strains showed various protein patterns among fusants. 4. From the dark culture experiment, growth of forage in inoculated soil with antagonistic bacteria was delayed than that of non-inoculated soil with antagonistic bacteria in each continuous cropping soil and in each sterilized soil. On the other hand, growth duration of forage was different between continuous cropping soil and sterilized soil. 5. Seed germination of Alfalfa, Italian ryegrass and Orchardgrass were significantly improved by inoculation of antagonistic bacteria(p< 0.05).

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Studies on Mulberry Shoot Rot caused by Fusarium spp. (Fusarium spp. 균에 의한 뽕나무신소썩음병에 관한 연구)

  • 윤형주;김영택;진경식;박인균;양성열
    • Journal of Sericultural and Entomological Science
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    • v.37 no.1
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    • pp.86-91
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    • 1995
  • Isolation and pathogenicity of Fusarium spp. from mulberry shoot rot and severity of diseases which were known as bacterial blight were examined on four mulberry varieties in Suwon, Kongju and Chuncheon, A symptom of mulberry shoot rot was initiated long brown spot on young leaves and shoots. It was developed into dark brown spot and produced white mycelia and spores on the diseased symptoms. A symptom of bacterial blight showed leaf rolling and water soaking spot and produced bacterial ooze on leaf and shoot However later stage of upper two types of symptom was hardly distinguished. Severities of shoot rot and bacterial blight were 7.5% and 4.4% in Suwon, respectively. Isolation of Fusarium spp. on shoot rot symptoms was highter than that on bacterial blight symptoms, but isolation of Pseudomonas spp. was higher on bacterial blight symptoms. Trends of pathogenicity of Fusarium spp. and Pseudomonas spp. were similar to inoculation works, and isolations of pathogenic Fusarium spp. from center of symptom was higher than that from 30cm of symptom of all samples in three cultivation areas. Disease severities of shoot rot on variety of Kaeryangppong were 13.9%, 15.9% and 17.2% in Suwon, Kongju and Chuncheon, respectively. However variety of Cheongolppong was highly resistant to shoot rot disease in three cultivation areas.

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Studies on Physico-chemical Properties an d Pathogenicity of Porcine Enterovirus Isolated from Feces of Pigherds (돼지 분변 유래 PORCINE ENTEROVIRUS의 물리화학적 특성 및 병원성에 관한 연구)

  • 박정우;이종인;신용호;조우영;최윤식
    • Korean Journal of Veterinary Service
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    • v.14 no.2
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    • pp.110-120
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    • 1991
  • 28 porcine enteroviruses were isolated from 86 pig-feces of 9 swine farms located in south region, Chung-buk, from March to September 1990. Physico-chemical properties and pathogenicity of isolates were investigated. Results obtained throughout experiments are summarized as follows. According to the age, weanlings(40-90 days), sucklings(10-30 days) and adult pigs(6 months over) showed the isolation rate of 67%. 8% and 4%, respectively. By physico-chemical tests, YD-90/22, YD-90/43 and YD-90/64 strains were found to be ether, chloroform and PH stable. Nucleic acid test suggests the virus to have a DNA genome. Most of the Isolates were not evident of hemagglutinin using erythrocytes from various mammalian & avian. 22 strains among the isolates were shown CPE type I and the remainders were CPE type II. 3 strains among isolates of CPE type I strains were neutralized with high titers to serotype 2 antiserum. In the study on virus growth curve in PK-l5 cells, YD-90/22, YD-90/43 and YD-90/64 strains showed the maximum infectivity titers($10^{6.0}-l0^{6.5} TCID({50}ml$) at 4days post inoculation(PI). When 30 day-old commercial piglets were inoculated only intraoral route with the YD-90/22 strain at $10^{6.0} TCID_{50}ml,$ piglets not showed the symptoms. But piglets inoculated by intramuscle route, intraoral and intramuscle route after pretreat with dexamethasone(2.5mg /kg) for 5 days were shown the symptoms of anorexia, diarrhea, pyrexia and ataxia at 4th-6th days PI. The viral reisolation in the virus-inoculated piglets was examined from feces. The viruses were recovered intermittently from 2nd to 16th day PI and at 4th-6th day PI, all piglets excreted viruses.

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Studies on the L-Glutamic acid Fermentation(Part I ) L-Glutamic acid Production from the Hydrolyzate of Sliced and Dried Sweet Potatoes (L-Gutamic acid희효생산에 관한 연구(제일보) 절간고구마 산분해액을 이용한 L-Glutamic acid 생산)

  • 양한철;최용진;양한우;성하진
    • Microbiology and Biotechnology Letters
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    • v.3 no.1
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    • pp.7-15
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    • 1975
  • The possibilities of utilizing acid-hydrolyzate of "Sliced and dried sweet potatoes" as a carbon source for the microbial production of L-Glutamic Acid(L-GA) with Micrococcus glutamicus were investigated and the results showed as follows: 1) The highest hydrolysis rate, 74.6% of the reducing sugar based on the weight of dry matter, was obtained when the sweet potatoes were hydrolyzed with 0.8% of HCI at 2.0kg/$cm^2$ for 30 minutes. The most favorable hydrolyzate for the growth of the cells, however, was found to be the one obtained by treating the sweet potatoes with 0.5% HCI at 2. 0kg/$cm^2$ for 10 minutes. Reducing sugar content of the hydrolyzate was 10% as glucose. 2) Biotin content of the hydrolyzate was 25$\mug$/1 and it was proved to be excess in amount for the L-GA production. 3) The effects of addition of antibiotics, alcohols and fatty acid esters on the L-GA production were tested in the biotin excess medium. The production of L-GA was most increased to 32.5g/l with the addition of 10 I. U. of penicillin per ml. to the culture medium at 4 hours after inoculation. But the addition of alcohols, especially fatty acid esters, showed no significant effects. 4) Among the organic nutrients tested. " Gluten acid hydrolyzate" greatly enhanced the production of L-GA adding it's concentration of 1.0% to the medium. 5) The maximum production of L-GA resulted in 35g/1 when the cells were grown for 48 hours in the hydrolyzate medium supplemented with 1.0% of "Gluten acid hydrolyfate" and with 10 I. U. of penicillin per ml added at 4 hours after cultivation.

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Studies on the supplementation of fermented soybean by Bacillus subtilis (natto) on performances, prevention against disease in broilers (낫도균을 이용한 닭 생산생과 질병예방에 관한 연구)

  • Kwon Mee-Soon;Lee Ji-Yoog;Park In-Gyu;Yoon Yeo-Baik;Joung Dong-Suk
    • Korean Journal of Veterinary Service
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    • v.29 no.3
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    • pp.257-266
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    • 2006
  • The present study was done to investigate the effect of dietary supplemental freezer dry powder fermentation soybean by Bacillus subtilis (natto) on the growth performance and intestinal microflora, prevention of fowl typhoid infection, the uptake of vegetative diet in broiler chickens. The chickens were fed control diet (supplement antibiotics) and fermentation soybean (0.75, 1.5, 3%) diets. A total of 280 one day old broiler chickens with randomly mixed sexes were fed the four diets for 6 weeks. Body weight gain of chicken fed 1.5% fermentation soybean by B subtilis tend to increase higher than the control from 6th week. Chickens fed diets containing 1.5% fermentation soybean by B subtilis had higher intake than those fed the other levels to the 3th weeks, but lower than control from the 4th week to the 6th week. Feed conversion also improved significantly in the supplemental 1.5% fermentation soybean by B subtilis from the 4th week to the 6th week. The number of B subtilis and Lactobacillus spp in the ileum and cecum tend to increase in the supplemental fermentation soybean by B subtilis at 6 week of age, but was not significantly different. In the nutrient digestibility, the feed conversion on the supplemental 1.5% fermentation soybean by B subtilis was better than the control and the weight of drying feces lower than the control. In test of S. gallinarum intramuscular inoculation, reisolation rate of S gallinarum in liver and feces 1.5% the fermentation soybean by B subtilis 75% (liver), 17% (feces) had decreased than the control.

Survival Studies of Some Enteric Pathogens in Sewage (장내 병원균의 하수중 생존 상태에 관한 연구)

  • 노일협;공인귀
    • YAKHAK HOEJI
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    • v.22 no.1
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    • pp.33-41
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    • 1978
  • In order to study the growth and survival of enteric pathogens causing water-borne infections in sewage, the filter-sterilized and autoclaved sewages of Dae Gu City were inoculated with Salmonella typhimuriuim, Shigella flexneri 2a, Sh. sonnei I, Vibrio eltor and V. parahaemolyticus, as test series and Escherichia coli as control. After varying periods of incubation up to 15 days at $4^{\circ}$, $15^{\circ}$, $25^{\circ}$ and $37^{\circ}C$, viable cells in the inoculated sewages were counted by colony count technique. Distilled water and 0.9% saline were subjected to inoculation of the organisms was observed in the filter-sterilized and autoclaved sewages at $4^{\circ}$ and the sewages became sterile within a few days. At $15^{\circ}$, no growth and rapid inactivation of the organisms in the filter-sterilized sewage and slight or no growth in the autoclaved sewage was noted. Some viable cells were found in the autoclaved sewage after 15 days. A considerable growth was observed in the filter-sterilized and autoclayed sewages, at $25^{\circ}$ and $37^{\circ}$, and large numbers of viable cells were found even after 15days of incubation. In general, the autoclaved sewage supproted the growth more noticeably than the filter-sterilized, except for V.parahaemolyticus which grew well in filter-sterilized sewage. No marked difference was noted between incubations at $25^{\circ}$ and $37^{\circ}$, but V. parahaemolyticus showed a slightly more active growth at $25^{\circ}$ than at $37^{\circ}$. Distilled water inactivated the organisms within a few days, but saline supported the growth at $25^{\circ}$ and $37^{\circ}$. Marked differences were noted in the survival test of sewages pathogens of different origins.

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Anticancer and Immunomodulatory Effects of Formosa Plum Extracts on Non-small Cell Lung Carcinoma, NCI-H520, Xenograft Mice (비소세포 폐암 세포 (NCI-H520) 이식 마우스에서 포모사 자두 추출물의 면역활성 및 악액질 억제효과)

  • Son, Young-Sook;Lee, Sang-nam;Park, Ji-Ha
    • The Korea Journal of Herbology
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    • v.35 no.6
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    • pp.35-41
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    • 2020
  • Objectives : The object of this study was to observe anticancer and immunomodulatory effects of Formosa plum aqueous extracts (PLe) on non-small cell lung carcinoma (squamous epithelial carcinoma), NCI-H520, xenograft Balb/c nu-nu nude mice. Method : Three different dosages of PLe, 50, 100 and 200 mg/kg were orally administered once a day for 28 days from 11 days after tumor cell inoculation. Five groups, each of seven mice per group were used in the present study as follows. Tumor volume and weights, serum interferon (IFN)-γ levels, serum IL-6 were observed with tumor mass and lymphatic organ histopathology to detect anticancer and immunomodulatory effects. Result : Although no meaningful changes on the tumor weights and volumes were observed after treatment of all three different dosages of PLe, decreases of tumor cell volumes in tumor masses were dose-dependently decreased mediated by increases of apoptosis among tumor cells by treatment of PLe 100 and 200 mg/kg as compared with tumor-bearing control. In addition, decreases on the body weight and gains were also demonstrated in tumor-bearing control with increases of serum IL-6 levels. Conclusion : The results obtained in this study suggest that over 50 mg/kg of PLe showed favorable immunomodulatory and anticachexic effects with anticancer effects in 100 and 200 mg/kg of PLe treated groups on the NCI-H520 cell xenograft. However, detail mechanism studies should be conducted in future with the screening of the biological active compounds in this herb.

Anticachexic Effects of Kwibi-tang on Non-small Cell Lung Carcinoma, NCI-H520 Xenograft Mice (귀비탕(歸脾湯)이 비소세포 폐암세포(NCI-H520) 이식 마우스에서 악액질 억제에 미치는 영향)

  • Son, Ji-young;Kim, Kyung-soon;Choi, Hae-Yun;Kim, Jong-Dae
    • Journal of Korean Traditional Oncology
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    • v.20 no.2
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    • pp.23-36
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    • 2015
  • Purpose : The object of this study was to observe anti-cachexic effects of Kwibi-tang extracts (KBTe) on non-small cell lung carcinoma (squamous epithelial carcinoma), NCI-H520, xenograft Balb/c nu-nu nude mice. Methods : Three different dosages of KBTe, 50, 100 and 200 mg/kg were orally administered once a day for 42 days from 11 days after tumor cell inoculation. Six groups, each of 8 mice per group were used in the present study. Changes on the body weight, the epididymal fat weight and serum IL-6 levels were detected with the thicknesses of deposited cervical brown adipose tissue and their mean diameters to monitor the tumor-related anticachexic effects. Results : Deceases on the body weight and gains were also demonstrated in tumor-bearing control with increases of serum IL-6 levels, decreases of epididymal fat pad weight, atrophic changes of cervical brown adipose tissues. These are means that tumor-related cachexia are induced by tumor cell inoculations in the present study. However, these tumor-related cachexia were markedly inhibited by all three different dosages of KBTe treatment as compared with tumor-bearing control. 5-FU showed somewhat deteriorated the tumor-related cachexia in the present study. Conclusion : The results obtained in this study suggest that over 50 mg/kg of KBTe showed favorable anticachexic effects on the NCI-H520 cell xenograft. However, detail mechanism studies should be conducted in future with the screening of the biological active compounds in this herb.

Electron Microscopic Study on the Replcation of Hantaan Virus in Vero-E6 Cells (Vero-E6 세포에서 한탄바이러스의 증식에 관한 전자현미경적 연구)

  • Park, Kyung-Hee;Seong, In-Wha
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.201-209
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    • 1999
  • Results of the studies on the morphologic and molecular biologic characteristics of Hantaan virus (HTNV), one of the etiologic agents of Hemorrhagic fever with renal syndrome (HFRS), revealed that HTNV was a member of Family Bunyaviridae and its RNA divided into three segments. And the nucleotide sequences of these segments also were known and the differences in nucleotide sequences of HTNV from other members of genus Hantavirus were clearly evaluated. But the morphorgenesis, pathogenesis of HFRS and the replication time had not been clearly determined. In this study, to estimate the replication time of HTNV in Vero E-6 cells, Vero cells were infected with HTNV 76/118 strain, and cells were harvested from two hours post-infection up to 24 hours at two hours-intervals. Harvested cells were treated with ordinary techniques for electron microscopy and immune-electron microscopy. And then thin sections were observed under transmission electron microscope. HTNV particles were not found in the cytoplasm and in the extracellular space between $2{\sim}8$ hours after inoculation of virus, but virus particles were observed in extracellular space near the cell membrane of Vero-E6 cells 10 hours after infection. In immune electron microscopy, mature HTNV particles in extracellular spaces and immature virus labelled with gold particles in the cytoplasm of Vero E-6 cell 10 hours after infection of HTNV could be seen. This results suggest that the replication time of HTNV might be about 10 hours.

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Platform of Hot Pepper Defense Genomics: Isolation of Pathogen Responsive Genes in Hot Pepper (Capsicum annuum L.) Non-Host Resistance Against Soybean Pustule Pathogen (Xanthomonas axonopodis pv. glycines)

  • Lee, Sang-Hyeob;Park, Do-Il
    • The Plant Pathology Journal
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    • v.20 no.1
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    • pp.46-51
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    • 2004
  • Host resistance is usually parasite-specific and is restricted to a particular pathogen races, and commonly is expressed against specific pathogen genotypes. In contrast, resistance shown by an entire plant species to a species of pathogen is known as non-host resistance. Therefore, non-host resistance is the more common and broad form of disease resistance exhibited by plants. As a first step to understand the mechanism of non-host plant defense, expressed sequence tags (EST) were generated from a hot pepper leaf cDNA library constructed from combined leaves collected at different time points after inoculation with non-host soybean pustule pathogen (Xanthomonas axonopodis pv. Glycines; Xag). To increase gene diversity, ESTs were also generated from cDNA libraries constructed from anthers and flower buds. Among a total of 10,061 ESTs, 8,525 were of sufficient quality to analyze further. Clustering analysis revealed that 55 % of all ESTs (4685) occurred only once. BLASTX analysis revealed that 74% of the ESTs had significant sequence similarity to known proteins present in the NCBI nr database. In addition, 1,265 ESTs were tentatively identified as being full-length cDNAs. Functional classification of the ESTs derived from pathogen-infected pepper leaves revealed that about 25% were disease- or defense-related genes. Furthermore, 323 (7%) ESTs were tentatively identified as being unique to hot pepper. This study represents the first analysis of sequence data from the hot pepper plant species. Although we focused on genes related to the plant defense response, our data will be useful for future comparative studies.