• Title/Summary/Keyword: strains inoculation

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Comparative Studies of Steel Wool Method and Gaspak Method for the Culture of Anaerobic Bacteria (염기성 세균 배양을 위한 Steel Wool법과 Gaspak법의 비교)

  • 정윤섭
    • Korean Journal of Microbiology
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    • v.12 no.3
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    • pp.131-137
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    • 1974
  • It is a well-known fact that an isolation of non-sporeforming anaerobes, considered normal flora in man ordinarily but causes serious infections sometimes, is a dificult procedure because of their great oxygen sensitivity. Among the many techniques employed in clinical laboratories, despite of its high expenses, the GasPak method has been most widely used because of its relative simplicity. On the other hand, the steel wool method has gained a good reputation recently. This technique makes it possible to treat individual plate so that any single specimen can be promptly cultured anaerobically. The procedure is quite simple and the expenses are negligible. In the present study it is to compare these two methods as to their efficiency of anaerobic cultivation using 13 VPI strains of non-sporeforming amaerobic bacteria. Among the 13 species the following 11, Bacteroides fragilis ss. fragilis, B. fragilis ss. thetaiotaomicron, propionibacterium acnes, Eubacterium limosum, E. lentum, peptococcus asaccharolyticus, Pc. prevotii, Pc. magnus, peptostreptococcus anaerobius, Ps. intermedius nad Veillonella parvula, grew well with the steel wool method whose colony numbers reaching 57 to 119% of those with GasPak method. The remaining two species, Fusobacterium nucleatum and F.necrophorum, did not grow well with the steel wool method showing the colony numbers were only 0.4% of those with GasPak method in the case of Fusobacterium nucleatum. In the case of Fusobacterium necrophorum, very few colonies developed even with a heavy inoculation. As to the size of colonies, there were no significant difference between these two methods.

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Bioavailability of slow-desorbable naphthalene in a biological air sparging system

  • Li, Guang-Chun;Chung, Seon-Yong;Park, Jeong-Hun
    • Advances in environmental research
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    • v.1 no.3
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    • pp.201-210
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    • 2012
  • The bioavailability of sorbed organic contaminants is one of the most important factors used to determine their fate in the environment. This study was conducted to evaluate the bioavailability of slow-desorbable naphthalene in soils. An air sparging system was utilized to remove dissolved (or desorbed) naphthalene continuously and to limit the bacterial utilization of dissolved naphthalene. A biological air sparging system (air sparging system with bacteria) was developed to evaluate the bioavailability of the slow-desorption fraction in soils. Three different strains (Pseudomonas putida G7, Pseudomonas sp. CZ6 and Burkholderia sp. KM1) and two soils were used. Slow-desorbable naphthalene continuously decreased under air sparging; however, a greater decrease was observed in response to the biological air sparging system. Enhanced bioavailability was not observed in the Jangseong soil. Overall, the results of this study suggests that the removal rate of slow-desorbable contaminants may be enhanced by inoculation of degrading bacteria into an air sparging system during the remediation of contaminated soils. However, the enhanced bioavailability was found to depend more on the soil properties than the bacterial characteristics.

The mycelial growth and ligninolytic enzyme activity of cauliflower mushroom (Sparassis latifolia)

  • Sou, Hong-Duck;Ryoo, Rhim;Ka, Kang-Hyeon;Park, Hyun
    • Forest Science and Technology
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    • v.13 no.4
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    • pp.158-163
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    • 2017
  • We examined the activities of lignin-degrading enzymes of the mycelium of cauliflower mushroom (Sparassis latifolia). Three different strains of S. latifolia collected from several sites in Korea and one crossbred strain were cultured on potato dextrose broth (PDB) and Kirk's medium in order to study the activities of their ligninolytic enzymes. Mycelial growth reached maximum levels between 14 and 21 days after inoculation and pH increased by 0.12 units over 35 days. Laccase activity began increasing after 14 days on both types of media. Manganese peroxidase (MnP) activity followed a trend similar to that of laccase on Kirk's medium, but not on PDB. The activity of lignin peroxidase (LiP) differed from that of other enzymes; its activity decreased by half after 14 days on PDB but remained constant on Kirk's medium over 35 days. The total protein concentration increased considerably after 14 days and peaked at 21 days on PDB. A similar maximum was attained on Kirk's medium. In contrast, the residual glucose increased rapidly at 14 days on Kirk's medium, while increasing gradually up to 28 days on PDB. This study indicates that S. latifolia is more similar to white rot fungi than to other brown rot fungi.

Ascospore Infection and Colletotrichum Species Causing Glomerella Leaf Spot of Apple in Uruguay

  • Alaniz, Sandra;Cuozzo, Vanessa;Martinez, Valentina;Stadnik, Marciel J.;Mondino, Pedro
    • The Plant Pathology Journal
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    • v.35 no.2
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    • pp.100-111
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    • 2019
  • Glomerella leaf spot (GLS) caused by Colletotrichum spp. is a destructive disease of apple restricted to a few regions worldwide. The distribution and evolution of GLS symptoms were observed for two years in Uruguay. The recurrent ascopore production on leaves and the widespread randomized distribution of symptoms throughout trees and orchard, suggest that ascospores play an important role in the disease dispersion. The ability of ascospores to produce typical GLS symptom was demonstrated by artificial inoculation. Colletotrichum strains causing GLS did not result in rot development, despite remaining alive in fruit lesions. Based on phylogenetic analysis of actin, ${\beta}$-tubulin and glyceraldehyde-3-phosphate dehydrogenase gene regions of 46 isolates, 25 from fruits and 21 from leaves, C. karstii was identified for the first time causing GLS in Uruguay and C. fructicola was found to be the most frequent (89%) and aggressive species. The higher aggressiveness of C. fructicola and its ability on to produce abundant fertile perithecia could help to explain the predominance of this species in the field.

Occurrence of Clubroot Caused by Plasmodiophora brassicae in Baecheongchae

  • Kim, Wan-Gyu;Oh, Sang-Keun;Semunyana, Marc;Han, Man-Jong;Lee, Gyo-Bin;Cho, Weon-Dae
    • The Korean Journal of Mycology
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    • v.48 no.4
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    • pp.499-503
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    • 2020
  • Clubroot symptoms were frequently observed on the roots of Baecheongchae plants grown in vinyl greenhouses of a farmer located in Yangpyeong area of Korea during a disease survey in June 2019. The incidence of diseased Baecheongchae plants ranged from 30 to 90% in the vinyl greenhouses investigated. Many resting spores were found in the tissue of root galls collected. The resting spores were hyaline and spherical and measured 2.5-4.2 ㎛ in diameter. Three inoculum suspensions of resting spores prepared from the root galls were inoculated to the roots of healthy Baecheongchae plants. All the inoculum suspensions caused clubroot symptoms to appear on the roots of the inoculated Baecheongchae plants. The symptoms on the roots induced by artificial inoculation were similar to those observed in the plants of the vinyl greenhouses during the disease survey. Resting spores of the pathogen were recovered from the root galls of the inoculated plants. Three root gall isolates obtained from the inoculated plants were used for molecular identification. Comparing the isolates to the Plasmodiophora brassicae strains in GenBank, the amplification products demonstrated 100% similarity with the internal transcribed spacer (ITS2) sequences. The clubroot pathogen was identified as P. brassicae according to its morphological, pathological, and molecular characteristics. This is the first report of P. brassicae causing clubroot in Baecheongchae.

First Report of Melon Soft Rot Disease Caused by Pectobacterium brasiliense in Korea

  • Kyoung-Taek Park;Leonid N. Ten;Chang-Gi Back;Soo-Min Hong;Seung-Yeol Lee;Jeung-Sul Han;Hee-Young Jung
    • Research in Plant Disease
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    • v.29 no.3
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    • pp.310-315
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    • 2023
  • In May 2021, characteristic soft rot symptoms, including soft, watery, slimy, black rot, wilting, and leaf collapse, were observed on melon plants (Cucumis melo) in Gokseong, Jeollanam-do, Korea. A bacterial strain, designated KNUB-06-21, was isolated from infected plant samples, taxonomically classified, and phylogenetically analyzed using 16S rRNA and housekeeping gene sequencing. Strain KNUB-06-21 was also examined for compound utilization using the API ID 32 GN system and strain KNUB-06-21 was identified as Pectobacterium brasiliense. Subsequent melon stem inoculation studies using strain KNUB-06-21 showed soft rot symptoms similar to field plants. Re-isolated strains shared phenotypic and molecular characteristics with the original P. brasiliense KNUB-06-21 strain. To our knowledge, ours is the first report of P. brasiliense causing melon soft rot disease in Korea.

First Report of Pectobacterium versatile as the Causal Pathogen of Soft Rot in Kimchi Cabbage in Korea

  • Kyoung-Taek Park;Soo-Min Hong;Chang-Gi Back;Young-Je Cho;Seung-Yeol Lee;Leonid N. Ten;Hee-Young Jung
    • Research in Plant Disease
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    • v.29 no.1
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    • pp.72-78
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    • 2023
  • In September 2021, gray-to-brown discoloration and expanding water-soaked lesions were observed on the outer and inner layers and the core of kimchi cabbage (Brassica rapa subsp. pekinensis) in fields located in Samcheok, Gangwondo, Korea. A bacterial strain designated as KNUB-02-21 was isolated from infected cabbage samples. Phylogenetic analysis based on the sequences of the 16S rRNA region and the dnaX, leuS, and recA genes confirmed that the strain was affiliated with Pectobacterium versatile. Additionally, the biochemical and morphological profiles of the isolate were similar to those of P. versatile. Based on these results, the isolate was identified as a novel strain of P. versatile. Healthy kimchi cabbage slices developed soft rot upon inoculation with P. versatile KNUB-02-21 and exhibited symptoms similar to those observed in the diseased plants in fields. The re-isolated strains were similar to those of P. versatile. Prior to our study, P. versatile as the causative pathogen of kimchi cabbage soft rot had not been reported in Korea.

The Antibacterial Properties of Filtrates from Chinese Cabbage Kimchi

  • Seong-Soo CHA;JeungSun LEE;Min-Kyu KWAK
    • The Korean Journal of Food & Health Convergence
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    • v.9 no.6
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    • pp.9-19
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    • 2023
  • Lactobacillus plantarum and Leuconostoc mesenteroides are crucial functional starters and predominant isolates in a wide range of fermented foods, particularly kimchi, whose constituents exhibit bioactive properties. We previously developed a methodology using anion exchange resins to purify peptidyl compounds from Lb. plantarum LBP-K10. Antibacterial cultures of Lb. plantarum LBP-K10 were obtained from the respective cultures' supernatants and filtrates. However, conclusive evidence of the efficacy of kimchi filtrates in eradicating pathogenic bacteria is lacking. We aimed to simulate the potential effects of antibacterial filtrates that contained antibacterial compounds which were derived from cultures of Lb. plantarum LBP-K10. We acquired the kimchi filtrates using a combination of centrifugation and filtration methodologies, without the requirement for inoculation. The filtered liquid from Chinese cabbage kimchi, inoculated with Lb. plantarum LBP-K10 as a starter culture, and the non-inoculated liquid from Chinese cabbage kimchi (referred to as CCK and CCKRef, respectively) were were examined. CCK demonstrated greater inhibitory activity and a more significant bactericidal effect against the bacterial indicator strains. The minimum inhibitory concentration demonstrated comparable outcomes in tests against both Gram-positive and Gram-negative bacteria. This research offers a groundbreaking examination that displays the effectiveness of profiling peptidyl compounds within kimchi filtrates for curing bacterial infections.

Effects of Cellulolytic Microbes Inoculation During Deep Stacking of Spent Mushroom Substrates on Cellulolytic Enzyme Activity and Nutrients Utilization by Sheep (버섯부산물 퇴적발효 시 섬유소 분해균 접종이 섬유소 분해성 효소 활력과 면양의 영양소 이용성에 미치는 영향)

  • Kim, Y.I.;Jun, S.H.;Yang, S.Y.;Huh, J.W;Kwak, W.S.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.667-676
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    • 2007
  • This study was conducted to determine effects of cellulolytic microbes inoculation to sawdust-based spent mushroom substrate(SMS) during deepstacking on fermentation parameters, total microbial counts and cellulolytic enzyme activity and to on SMS nutrients utilization by sheep. For sheep metabolism trials, six sheep(ram, average 54.8kg) were fed a Control diet(70% concentrates, 15% rice straw and 15% SMS with no microbial treatment on a dry basis) and a Treatment diet(the same diet including SMS with a microbial treatment) for 2 trials. Spent mushroom substrates with or without a microbial(4 strains including 1 strain of Enterobacter ludwigii, 1 strain of Bacillus cereus and 2 strains of Bacillus subtillis) treatment (1% of SMS on wet basis) were deepstacked for 7 days. The internal temperatures in 1.2 M/T of SMS deepstacks reached to 50±5℃ within 7 days of storage. Total microbial counts remarkably decreased (P<0.05) with a deepstacking process and were not affected(P>0.05) by the microbial treatment. For fibrolytic enzyme activity, CMCase and xylanase activities were decreased(P<0.05) by a deepstacking process. After deepstacking, the microbial treatment showed about 2.5-times higher(P<0.05) for CMCase activity and about 4-times higher(P<0.05) for xylanase activity than those of the Control. Activities of ligninolytic enzymes such as laccase and MnP were not affected by the microbial treatment. The sheep fed the microbially treated SMS diet had a tendency of greater total tract digestibilities of ash(P=0.051), NFE (P=0.071), hemicellulose(P=0.087) and NDF(P=0.096) than those fed the untreated SMS diet. Nitrogen balance of sheep was not affected(P>0.05) by feeding of microbially treated SMS. Accordingly, these results indicate that cellulolytic microbes inoculation during deepstacking of SMS may improve the bio- utilization of SMS by sheep.

Effects of Pseudomonas Fluorescens, KR-164 on Plant Pathogenic Microorganisms (식물(植物) 병원성(病源性) 미생물(微生物)에 미치는 Pseudomonas fluorescens, KR-164의 영향(影響))

  • Rhee, Young-Hwan;Kim, Yeong-Yil;Lee, Jae-Pyeong;Kim, Yong-Wong;Kim, Yong-Jae;Lee, Jae-Wha
    • Korean Journal of Soil Science and Fertilizer
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    • v.23 no.1
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    • pp.53-59
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    • 1990
  • The antagonistic fluorescent pseudomonas, which was isolated from continuous cropping rhizosphere of pepper and cucumber, was identified as Pseudomonas fluorescens (P.f.). For further study, transformant was derived from the isolated P.f. after spontaneous mutation to give antibiotic resistance to nalidixic acid and rifampicin as marked strain. Both P.f. and transformant strains were used for this study and the results obtained were summarized as follows. 1. One of the most effective antagonistic strain, KR164, was selected against F. solani, F. oxysporum, R. solani and this strain was identified and classified as Pseudomonas fluorescens biotype IV. 2. Transformant, KR1641, was derived from strain KR164 and both strains had the same biological and biochemical characteristics. 3, Mycelial lysis and abnormal mycelia of plant pathogenic fungi were microscopically observed after simultaneous culture of fungus and given bacterial strain. 4. The length of chinese cabbage to the autolyzed became longer with given bacterial strain in dark culture. 5. Percentage of germination, number of leaves, length of height, and length of root in chinese cabbage in pot experiment were improved by inoculation of given bacterial strain. 6. The number of given bacterial strain kept generally stable until 34 days after inoculation of itself in pot experiment. Inoculation of given bacterial strain did affect the number of plant disease fungi to be decreased but did not affect the number of other bacteria, Bacillus, in pot experiment.

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