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Bacillus ginsengihumi sp. nov., a Novel Species Isolated from Soil of a Ginseng Field in Pocheon Province, South Korea

  • Ten Leonid N.;Im Wan-Taek;Baek Sang-Hoon;Lee, Jung-Sook;Oh, Hee-Mock;Lee, Sung-Taik
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1554-1560
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    • 2006
  • A Gram-positive, aerobic or facultative anaerobic, non motile, endospore-forming bacterial strain, designated Gsoil $114^T$, was isolated from a soil sample of a ginseng field in Pocheon Province (South Korea), and was characterized taxonomically by using a polyphasic approach. It grew well on nutrient agar medium and utilized a limited number of organic substrates as sole carbon sources, including D-xylose and some other carbohydrates, but did not utilize L-amino acids and organic acids. The isolate was positive for oxidase test but negative for catalase, and negative for degradation of macromolecules such as starch, cellulose, xylan, casein, chitin, and DNA. The G+C content of the genomic DNA was 41.8 mol%. The predominant isoprenoid quinone was menaquinone 7 (MK-7). The major fatty acids were $anteiso-C_{15:0}$ (32.1%), $iso-C_{15:0}$ (30.5%), and $anteiso-C_{17:0}$ (30.2%). Comparative 16S rRNA gene sequence analysis showed that strain Gsoil $114^T$ fell within the radiation of the cluster comprising Bacillus species and joined Bacillus shackletonii LMG $18435^T$ with a bootstrap value of 95%. The highest 16S rRNA gene sequence similarities were found with Bacillus shackletonii LMG $18435^T$ (97.6%), Bacillus acidicola DSM $14745^T$ (96.9%), Bacillus sporothermodurans DSM $10599^T$ (96.5%), and Bacillus oleronius DSM $9356^T$ (96.5%). The phylogenetic distance from any other validly described species within the genus Bacillus was less than 96%. DNA-DNA hybridization experiments showed that the DNA-similarities between strain Gsoil $114^T$ and closest phylogenetic neighbors were less than 39%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil $114^T$ (=KCTC $13944^T$=DSMZ $18134^T$) was classified in the genus Bacillus as the type strain of a novel species, for which the name Bacillus ginsengihumi sp. nov. is proposed.

ELISA Development for the residue of the organophosphorus insecticide acephate (ELISA에 의한 유기인계 살충제 Acephate 잔류물 분석법 개발)

  • Lee, Jae-Koo;Ahn, Ki-Chang;Stoutamire, Donald W.;Gee, Shirley J.;Hammock, Bruce D.
    • The Korean Journal of Pesticide Science
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    • v.5 no.2
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    • pp.1-12
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    • 2001
  • A competitive indirect enzyme-linked immunosorbent assay (ci ELISA) for the organophosphorus insecticide acephate, O,S-dimethyl acetylphosphoramidothioate, was developed using a polyclonal antibody. Three different haptens mimicking the analyze and containing hexanoic acid moiety as a linker were synthesized, and then conjugated with the carrier proteins bovine serum albumin and keyhole limpet hemocyanin by the N-hydroxysuccinimide active ester method. Polyclonal antibodies raised against hapten-KLH conjugates in rabbits and the hapten-BSA conjugates as coating antigens were screened and selected for the assay in the homologous and/or heterologous ELISA system. The effects of various assay conditions, including blocking reagents, detergent content, organic solvents, pH, and preincubation of tile mixture of the polyclonal antibody and the analyze on the sensitivity were evaluated. The $IC_{50}$ value of acephate of 110 ng/mL was obtained in an optimized heterologous system using hapten-3-BSA as a coating antigen and a polyclonal antibody 8377, showing the detection range of 10-1000 ng/mL and the lowest detection limit of 4 ng/mL. The cross-reactivities of the structurally related insecticides, including methamidophos were less than 0.02%. These results indicate that the ELISA could be a convenient and alternative tool for monitoring acephate residues in agricultural products and environmental samples.

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Saprolegnia parasitica Isolated from Rainbow Trout in Korea: Characterization, Anti-Saprolegnia Activity and Host Pathogen Interaction in Zebrafish Disease Model

  • Shin, Sangyeop;Kulatunga, D.C.M.;Dananjaya, S.H.S.;Nikapitiya, Chamilani;Lee, Jehee;De Zoysa, Mahanama
    • Mycobiology
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    • v.45 no.4
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    • pp.297-311
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    • 2017
  • Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin $[IL]-1{\beta}$, tumor necrosis factor ${\alpha}$, IL-6, IL-8, interferon ${\gamma}$, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules ($CD8^+$ and $CD4^+$) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as $200{\mu}g/mL$ and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.

Production of Phytol, an ACAT Inhibitor, from Callus Culture of Lettuce (Lactuca sativa L.) (상추 (Lactuca sativa L.) callus로부터 ACAT 억제 활성물질, phytol의 생산)

  • An, Kwang-Hee;Jang, Tae-O;Baek, Nam-In;Kim, Se-Young
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.63-68
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    • 2002
  • The possibility for mass production of phytol, inhibitory diterpene against ACAT (Acyl-CoA: Cholesterol acyltransferase) was investigated by using callus culture of lettuce. The callus were induced from lettuce cotyledon explants on MS medium containing 0.5 mg.L$^{-1}$ NAA after 4 week's culture. Adventitious roots were formed from the explants on MS medium containing 0.5 mg.L$^{-1}$ IBA or 1.0 mg.L$^{-1}$ NAA. Adventitious shoots and roots were emerged from the callus when the callus was transferred to MS medium containing auxin alone, or with cytokinin. The plant growth regulators and their concentrations for the organogenesis were 1.0 mg.L$^{-1}$ NAA, 0.1 mg.L$^{-1}$ BA, 0.5 mg.L$^{-1}$ NAA with 0.1 mg.L$^{-1}$ kinetin, or 0.5 g.L$^{-1}$ 2.4-D with 1.0 mg.L$^{-1}$ kinetin. Analyses of chlorophyll contents showed that chlorophyll contents were higher in morphogenic calli than in non-morphogenic calli. However, the chemical analyses of gas chromatography indicated that phytol contents were not proportionate to the chlorophyll contents of callus. The content of phytol was higher in callus than in lettuce cotyledon.ledon.

Effects of Posttraumatic Hypothermia in an Animal Model of Traumatic Brain Injury(TBI) - Immunohistochemical Stain by TUNEL & β-APP - (실험적 외상성 뇌손상모델에서 외상 후 저체온법의 효과 - TUNEL과 β-APP Immunohistochemical Stain -)

  • An, Byeong Kil;Ha, Young Soo;Hyun, Dong Keun;Park, Chong Oon;Kim, Joon Mee
    • Journal of Korean Neurosurgical Society
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    • v.29 no.4
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    • pp.461-470
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    • 2000
  • Objective : Many investigators have demonstrated the protective effects of hypothermia following traumatic brain injury(TBI) in both animals and humans. It has long been recognized that mild to moderate hypothermia improves neurologic outcomes as well as reduces histologic and biochemical sequelae after TBI. In this study, two immunohistochemical staining using terminal deoxynucleotidyl-transferase-mediated biotin dUTP nick end labeling(TUNEL), staining of apoptosis, and ${\beta}$-amyloid precursor protein(${\beta}$-APP), a marker of axonal injury, were done and the authors evaluated the protective effects of hypothermia on axonal and neuronal injury after TBI in rats. Material and Method : The animals were prepared for the delivery of impact-acceleration brain injury as described by Marmarou and colleagues. TBI is achieved by allowing of a weight drop of 450gm, 1 m height to fall onto a metallic disc fixed on the intact skull of the rats. Fourty Sprague-Dawley rats weighing 400 to 450g were subjected to experimental TBI induced by an impact-acceleration device. Twenty rats were subjected to hypothermia after injury, with their rectal temperatures maintained at $32^{\circ}C$ for 1 hour. After this 1-hour period of hypothermia, rewarming to normothermic levels was accomplished over 30-minute period. Following 12 hours, 24 hours, 1 week and 2 weeks later the animals were killed and semiserial sagittal sections of the brain were reacted for visualization of the apoptosis and ${\beta}$-APP. Results : The density of ${\beta}$-APP marked damaged axons within the corticospinal tract at the pontomedullary junction and apoptotic cells at the contused cerebral cortex were calculated for each animal. In comparison with the untreated controls, a significant reduction in ${\beta}$-APP marked damaged axonal density and apoptotic cells were found in all hypothermic animals(p<0.05). Conclusion : This study shows that the posttraumatic hypothermia result in substantial protection in TBI, at least in terms of the injured axons and neurons.

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Korean Red Ginseng-intake has Definite Clinical Usefulness and causes Nef Gene Variation including High Frequency of Deletion

  • Cho Young Keol;Lee Hee Kyung;Ahn Sun Hee;Lee Hee Jung;Nam Ki Yeul
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.185-211
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    • 2002
  • We have found many beneficial effects of the long-tenn intake of Korean red ginseng (KRG) in human immunodeficiency virus (HIV) type-I infected patients, including the maintenance of CD4+ T cell count for 10 years with KRG only and the delayed development of resistance mutation to ZDV. In this study, to investigate whether KRG-intake could affect the clinical progression and nef gene variation, we determined 200nef sequences from 70 patients. Follow-up period was $8.8{\pm}2.9$ years and annual decrease in CD4+T cell was $41{\pm}57/ul.$ Nested polymerase chain reaction (PCR) and direct sequencing were perfonned with peripheral blood mononuclear cells (PBMC) obtained at times during the study period. First, there was a significant correlation between survival duration and duration of KRG-intake $(36.8{\pm}38$ months)(P=0.000). There were significant correlations between the last NefProg score and CD4+ T cell count (r= 0.208, P<0.05) and annual decrease in CD4+ T cell count (r =0.346, P<0.01) in 70 patients. In addition, there were significant correlations between KRG-intake and annual decrease (r= 0.323, P<0.01), and the CD4+ T cell count itself (r=0.229, p<0.05). Furthennore, there was also a mild significance between the NefProg score and the duration of KRG-intake in only SP and RP (n=30, r=-0.281, P=0.067). In addition, we detected various defects in 21 patients $(30.0\%),$ not including 5 premature stop codons. Ten $(12.5\%)$ patients showed repeated deletion of an amino acid. Four of 10 patients were gross deletions and they were treated with KRG for more than 20 months. The number of patients with repeated gross deletions was significantly higher in the order of slow progressors $(18\%)$, typical progressors($3\%$), and rapid progressors($0\%$) (P<0.05). We also observed that long-tenn intake of KRG might make the change from A or D to T at position 54 and decrease NefProg score. Taken together, our results show clear evidence that the long-term intake of KRG has effects on nef gene variation as well as definite clinical usefulness.

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Application of lactic acid bacteria producing antifungal substance and carboxylesterase on whole crop rice silage with different dry matter

  • Lee, Seong Shin;Paradhipta, Dimas Hand Vidya;Lee, Hyuk Jun;Joo, Young Ho;Noh, Hyeon Tak;Choi, Jeong Seok;Ji, Keum Bae;Kim, Sam Churl
    • Animal Bioscience
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    • v.34 no.6
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    • pp.1029-1037
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    • 2021
  • Objective: This study was conducted to investigate effects of antifungal substance and carboxylesterase-producing inoculant on fermentation indices and rumen degradation kinetics of whole crop rice (WCR) silage ensiled at different dry matter (DM) contents. Methods: Dual-purpose inoculants, Lactobacillus brevis 5M2 and Lactobacillus buchneri 6M1, confirmed both activities of antifungal and carboxylesterase in the previous study. The WCR at mature stage was chopped, and then wilted to obtain three different DM contents consisting of 35.4%, 43.6%, and 51.5%. All WCR forages were applied distilled water (CON) or mixed inoculants with 1:1 ratio at 1×105 colony forming unit/g (INO), and ensiled into 20 L mini silo (5 kg) in quadruplicates for 108 d. Results: The INO silages had lower lactate (p<0.001) and butyrate (p = 0.022) with higher acetate (p<0.001) and propionate (p<0.001) than those of CON silages. Ammonia-N (p<0.001), lactate (tendency; p = 0.068), acetate (p = 0.030), and butyrate (p<0.001) concentrations of INO silages decreased linearly with increasing DM content of WCR forage. The INO silages presented higher lactic acid bacteria (p<0.001) with lower molds (p<0.001) than those of CON silages. Yeasts (p = 0.042) and molds (p = 0.046) of WCR silages decreased linearly with increasing DM content of WCR forage. In the rumen, INO silages had higher the total degradable fraction (p<0.001), total volatile fatty acid (tendency; p = 0.097), and acetate (p = 0.007), but lower the fractional degradation rate (p = 0.011) and propionate (p<0.001) than those of CON silage. The total degradable fraction (p<0.001), total volatile fatty acid (p = 0.001), iso-butyrate (p = 0.036), and valerate (p = 0.008) decreased linearly with increasing DM content of WCR forage, while the lag phase (p<0.001) was increased linearly. Conclusion: This study concluded that application of dual-purpose inoculants on WCR silage confirmed antifungal and carboxylesterase activities by inhibiting mold and improving rumen digestibility, while increase of wilting times decreased organic acids production and rumen digestibility.

A New Pink Mini Cut Flower Gerbera Cultivar, "Summer Ring" (핑크색 소륜 절화용 다수성 거베라 신품종 "썸머링")

  • Chung, Yong-Mo;Hwang, Ju-Chean;Chin, Young-Don;Kim, Su-Kyeong;Yi, Young-Byung;Kwon, Oh-Chang
    • Korean Journal of Breeding Science
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    • v.40 no.1
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    • pp.80-83
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    • 2008
  • A new pink gerbera (Gerbera hybrida Hort.) cultivar 'Summer Ring' was developed from a cross between 'Grandeur' and 'Nova Zembla' followed by seedling and line selections at the Flower Breeding Research Institute, Gyeongnam Agricultural Research and Extension Services (ARES) in 2005. Characteristics of the new cultivar were evaluated three times from 2003 to 2004. 'Summer Ring' has semi-double type mini flower in good harmony with pink (RHS, 56-D) ray floret and green center. It has good stable flower shape and strong peduncle, and its vase life was 14.7 days. The average yield of 'Summer Ring' flower was 69 per plant/year in greenhouse yield trails during 2003 and 2004. This cultivar was registered for commercialization in 2005. Commercial production of 'Summer Ring' could be possible throughout the country under greenhouse condition.

Growth performance, carcass and meat quality of lambs supplemented different vegetable oils

  • Miltko, Renata;Majewska, Malgorzata Paulina;Belzecki, Grzegorz;Kula, Katarzyna;Kowalik, Barbara
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.767-775
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    • 2019
  • Objective: The purpose of this study was to evaluate the effect of rapeseed and linseed oil supplementations on performance and meat quality of lambs. Methods: The experiment was conducted on 18 growing (100-day-old) lambs of $19.7{\pm}1.9kg$ live weight, assigned to 3 groups of 6 animals each. Control lambs were fed meadow hay and concentrate alone. Experimental animals additionally received rapeseed or linseed oils at a dose of 50 g/d. The lambs were slaughtered at an average body weight of $35.7{\pm}0.5kg$. Results: The dressing percentage was higher in lambs fed rapeseed oil. Total saturated fatty acids (SFA) and C15:0, C16:0, C17:0, C21:0, C24:0 were lower in longissimus dorsi muscle (MLD) in lambs fed linseed oil. Supplementation of diet with linseed oil decreased concentrations of total monounsaturated fatty acids and C16:1, C17:1, C18:1 cis-9 in MLD. The concentrations of n-3 polyunsaturated fatty acids (PUFA) and C18:3 n-3, C20:5 n-3 in MLD were higher in lambs fed linseed oil than in other groups. Oils supplementation to diets resulted in increased concentration of C22:6 n-3 in MLD. The inclusion of linseed oil into the diet increased the contents of total PUFA, n-3 PUFA and C18:3 n-3, C20:5 n-3, C22:6 n-3 in semitendinosus muscle in comparison to control. A tendency towards a lower n:6/n:3 ratio in MLD was observed when lambs were supplemented linseed oil. Conclusion: The supplementation of linseed oil to diets seems to reduce the concentration of SFA and increase the concentration of n-3 PUFA. The n-6/n-3 ratio is an important nutritional factor, and its value has been favorably decreased below 2, thereby achieving an important target related to human health. Due to these changes carcass fatty acid profile was improved, and so enhanced lamb meat healthy properties.

Changes in fermentation pattern and quality of Italian ryegrass (Lolium multiflorum Lam.) silage by wilting and inoculant treatments

  • Liu, Chang;Zhao, Guo Qiang;Wei, Sheng Nan;Kim, Hak Jin;Li, Yan Fen;Kim, Jong Geun
    • Animal Bioscience
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    • v.34 no.1
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    • pp.48-55
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    • 2021
  • Objective: This study was conducted to investigate the effects of wilting and microbial inoculant treatment on the fermentation pattern and quality of Italian ryegrass silage. Methods: Italian ryegrass was harvested at heading stage and ensiled into vinyl bags (20 cm×30 cm) for 60d. Italian ryegrass was ensiled with 4 treatments (NWNA, no-wilting noadditive; NWA, no-wilting with additive; WNA, wilting no-additive; WA, wilting with additive) in 3 replications, wilting time was 5 hours and additives were treated with 106 cfu/g of Lactobacillus plantarum. The silages samples were collected at 1, 2, 3, 5, 10, 20, 30, 45, and 60 days after ensiling and analyzed for the ensiling quality and characteristics of fermentation patterns. Results: Wilting treatment resulted in lower crude protein and in vitro dry matter digestibility and there were no significant differences in acid detergent fiber (ADF), total digestible nutrient (TDN), water-soluble carbohydrate (WSC), ammonia content, and pH (p>0.05). However, wilting treatment resulted in higher ADF and neutral detergent fiber content of Italian ryegrass silage (p<0.05), and the WNA treatment showed the lowest TDN and in vitro dry matter digestibility. The pH of the silage was higher in the wilting group (WNA and WA) and lower in the additive treatment group. Meanwhile, the decrease in pH occurred sharply between the 3-5th day of storage. The ammonia nitrogen content was significantly lower in the additive treatment (p<0.05), and wilting had no effect. As fermentation progressed, the lactic and acetic acid contents were increased and showed the highest content at 30 days of storage. Conclusion: The wilting treatment did not significantly improve the silage fermentation, but the inoculant treatment improved the fermentation patterns and quality of the silage. So, inoculation before ensiling is recommended when preparing high quality of Italian ryegrass silage, and when wilting, it is recommended to combine inoculation for making high quality silage.