• Title/Summary/Keyword: nutrient agar plate

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Fibrinolytic Activity and Proteomic Analysis of Bacillus licheniformis HK-12 Isolated from Chungkuk-Jang (청국장에서 분리한 Bacillus licheniformis HK-12의 혈전용해활성과 프로테옴 분석)

  • Sohn, Byung-Hee;Kwon, Sang-Chul;Oh, Kye-Heon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.9 no.3
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    • pp.800-806
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    • 2008
  • The strain HK-12 was enriched and isolated from naturally fermented soybean for the production of fibrinolytic enzyme and the proteome of this enzyme induced during the incubation period was analyzed. The activity of fibrinolytic enzyme derived from supernatants of the HK-12 culture was performed by fibrin plate method for solid fibrinolytic activity. As the result, the fibrinolytic activity of HK-12 grown on the nutrient agar media was about 2.3 times greater than that of plasmin used as standard. The purified enzyme was prepared by a series of purification process including ammonium sulfate precipitation, DEAE-cellulose, Sephadex chromatography. The molecular weight of the enzyme was determined to approximately 23kDa with SDS-PAGE. In order to examine which strain HK-12 proteins increased or decreased during the incubation period, 2-DE analysis was performed. Protein spot #1 significantly expressed on the 2-DE gel of bacteria cultivated for 36-hrs was analysed. As the result of protein sequence analysis using MALDI-TOF MS, one protein was identified as serine protein kinase (PrkA).

Characterization of Viable But Nonculturable Condition of Escherichia coli Induced with Copper (구리에 의해 유도된 VBNC 대장균의 특성)

  • Ku, Hyung-Keun;Park, Sang-Ryoul;Kim, Sook-Kyung
    • Microbiology and Biotechnology Letters
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    • v.36 no.3
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    • pp.209-214
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    • 2008
  • VBNC (Viable but nonculturable) state is an adaptive response of cells in adverse environments, which lead cell not grow on routine nutrient agar. In this study, we induced VBNC in Escherichia coli using copper and verify the characterization of it. After treatment of copper, we didn't detect any cells via plate cultivation, namely, colony forming unit (CFU) was zero. However, we identified the existence of VBNC by staining live cells with Live/Dead BacLight bacterial viability kit and counting them through flow cytometry. Then we isolated genomic DNA and RNA from VBNC-induced cells and analyzed the stability of them. Degradation of RNA is more severe than that of DNA and RNA is degraded as specific fragments. In addition, we showed the morphology of VBNC cell by Bio-Transmission Electron Microscope (Bio-TEM). VBNC cell showed impaired periplasmic space and inner and outer membrane were separated and the amount of cytosol were significantly decreased.

A Study on the Development of Microorganism Test Strips (대장균군 검사용 간이 시험지 개발)

  • 이인애;김재화;이희구;성창근;최인성;정태화
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.49-55
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    • 1996
  • The objective of this study was to develop a paper strip which could determine E. coli qualitatively and quantitatively in water, wastewater, drinks, or food. This paper strip method was a simple and rapid test method that determine E. coli by visual identification. In this study, nutrient culture media were formulated and characterized for optimum conditions. Paper strips were then prepared by impregnating into the media and dried at $60^\circ$C. The test procedure is quite simple to use. The paper strip was dipped into a sample, and excess sample was removed. The strip was then incubated at $37^\circ$C for 16 to 20 hours and the number of colonies on the strip was counted. The color of the colony spots produced by microorganisms varied depending on the media formulation. Violet-red spots were produced by E. coli. The test method was simple, rapid and no special laboratory equipment was necessary for visual identification. Therefore, this test method is applicable to on-site tests such as field tests or home tests. The paper strip method was compared with the standard agar plate method and Japanese commercial product. The method of the economical preparation of test strips was studied for production on industrial scale.

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Antimicrobial Characteristics of Different Parts of Guava against Food-Borne Bacteria (식중독 세균에 대한 구아바 부위별 추출물의 항균 특성)

  • Jo, Young-Hong;Ok, Dul-lee;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.12
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    • pp.1773-1778
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    • 2009
  • Guava (Psidium guajava L.) contains high amount of vitamins and minerals, and its leaves have been reported to be very effective on reducing blood pressure. In this study, antimicrobial characteristics of extracts from four different parts of guava (fruit, branch, leaf, and seed) with four different solvents (methanol, ethanol, acetone, and water) were evaluated. Four targeted food-borne microorganisms were selected; two Gram negatives (Escherichia coli O157:H7 and Salmonella Typhimurium) and two Gram positives (Staphylococcus aureus and Listeria monocytogenes). By the paper disc method, guava extracts showed stronger clear hollow zone against Gram positive bacteria than Gram negatives in nutrient broth agar plate. Especially, extracts of branches and leaves showed significant antibacterial activity. Guava extracts also showed significant inhibition activity on the growth of Gram positive food-borne bacteria in nutrient broth. For example, S. aureus did not grow at all at 200 ppm of acetone extracts of guava branch and leaf. In the case of L. monocytogenes, the same concentration of acetone extracts of guava branch and leaf inhibited the growth 33.5% and 55.4%, respectively, at 32 hrs of incubation time. The results indicated that extracts of guava branch and leaf showed significant antibacterial activities against food-borne Gram positive microorganisms, and that guava branches, the byproducts of guava, might be a valuable resource for antibacterial materials.

Study on CsRCI2D and CsRCI2H for improvement of abiotic stress tolerance in Camelina sativa L.

  • Lim, Hyun-Gyu;Kim, Hyun-Sung;Kim, Jung-Eun;Ahn, Sung-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.196-196
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    • 2017
  • Oilseed crop Camelina (Camelina sativa L.) is a suitable for biodiesel production that has high adaptability under low-nutrient condition like marginal land and requires low-input cost for cultivation. Enhanced abiotic stress tolerance of Camelina is very important for oil production under the wide range of different climate. CsRCI2s (Rare Cold Inducible 2) are related proteins in various abiotic stresses that predicted to localized at plasma membrane (PM) and endoplasmic reticulum (ER). These proteins are consist of eight-family that can be divided into tail (CsRCI2D/E/F/G) and no-tail (CsRCI2A/B/E/H) type of C-terminal. However, it is still less understood the function of C-terminal tail. In this study, CsRCI2D/H genes were cloned through gateway cloning system that used pCB302-3 as destination vector. And we used agrobacterium-mediated transformation system for generation of overexpression (OX) transformants. Overexpression of target gene was confirmed using RT-PCR and segregation ratio on selection media. We analyzed physiological response in media and soil under abiotic stresses using CsRCI2D and CsRCI2H overexpression plant. To compare abiotic stresses tolerance, wild type and CsRCI2D/H OX line seeds were sown on agar plate treated with various NaCl and mannitol concentration for 7 days. In the test of growth rate under abiotic stress on media, CsRCI2H OX line showed similar to NaCl and mannitol stress. In the other hand, CsRCI2D OX line showed to be improved stress tolerance that especially increased in 200mM NaCl but was similar on mannitol media. In greenhouse, WT and CsRCI2D/H OX lines for physiological analysis and productivity under abiotic stresses were treated 100, 150, 200mM NaCl. Then it was measured various parameters such as leaf width and length, plant height, total seed weight, flower number, seed number. CsRCI2H OX line in greenhouse did not show any changes in physiological parameters but CsRCI2D OX line was improved both physiological response and productivity under NaCl stress. Among physiological parameters of CsRCI2D OX line under NaCl stress, leaf length and width were observed shorter than WT but it were slightly longer than WT in 200mM NaCl stress. Furthermore, total seed weight of CsRCI2D OX line under stress displayed to decrease than WT in normal condition, but it was gradually raised with increasing NaCl stress then more than WT relatively. These results suggested CsRCI2D might be contribute to improve abiotic stress tolerance. However, function of CsRCI2H is need to more detail study. In conclusion, overexpression of CsRCI2s family can generate various environmental stress tolerance plant and may improve crop productivity for bio-energy production.

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PHOTOCATALYTIC ANTIEUNGAL ACTIVITY AGAINST CANDIDA ALBICANS BY $TiO_2$ COATED ACRYLIC RESIN DENTURE BASE

  • Yang Ji-Yeon;Kim Hee-Jung;Chung Chae-Heon
    • The Journal of Korean Academy of Prosthodontics
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    • v.44 no.3
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    • pp.284-294
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    • 2006
  • Statement of problem. Proliferation of Candida albicans is primarily within the plaque on the fitting surface of the denture rather than on the inflamed mucosa. Consequently, the treatment of the denture is equally important as treatment of the tissue. Cleansing and disinfection should be efficiently carried-out as the organisms can penetrate into the voids of the acrylic resin and grow in them, from which they can continue to infect and reinfect bearing tissues. Purpose. The purpose of this study was to evaluate the applicability of photocatalytic reaction to eliminate Candida albicans from acrylic resin denture base, and to investigate the anti-fungal effect with various UVA illumination time. Materials and Methods. The specimens were cured by the conventional method following the manufacturer's instruction using thermal polymerized denture base resin (Vertex RS: Dentimex, Netherlands). $TiO_2$ photocatalyst sol(LT), which is able to be coated at normal temperature, was made from the Ti-alkoxide progenitor. The XRD patterns, TEM images and nitrogen absorption ability of the $TiO_2$ photocatalyst sol(LT) were compared with the commercial $TiO_2$ photocatalyst P-25. The experimental specimens were coated with the mixture of the $TiO_2$ photocatalyst sol(LT) and binder material (silane) using dip-coater, and uncoated resin plates were used as the control group. Crystallinity of $TiO_2$ of the specimen was tested by the XRD. Size, shape and chemical compositions were also analyzed using the FE-SEM/ EDS. The angle and methylene blue degradation efsciency were measured for evaluating the photocatalytic activity of the $TiO_2$ film. Finally, the antifungal activity of the specimen was tested. Candida albicans KCTC 7629(1 ml, initial concentration $10^5$ cells/ ml) were applied to the experiment and control group specimens and subsequently two UVA light source with 10W, 353 nm peak emission were illuminated to the specimens from 15cm above. The extracted $2{\mu}l$ of sample was plated on nutrient agar plate ($Bacto^{TM}$ Brain Heart Infusion; BD, USA) with 10 minute intervals for 120 minute, respectively. It was incubated for 24 hours at $37^{\circ}C$ and the colony forming units (CFUs) were then counted. Results. Compared the characteristics of LT photocatalyst with commercial P-25 photocatalyst, LT were shown higher activity than P-25. The LT coated experimental specimen surface had anatase crystal form, less than 20 nm of particle size and wide specific surface area. To evaluate the photocatalytic activity of specimens, methylene blue degradation reaction were used and about 5% of degradation rate were measured after 2 hours. The average contact angle was less than $20^{\circ}$ indicating that the LT photocatalyst had hydrophilicity. In the antifungal activity test for Candida albicans, 0% survival rate were measured within 30 minute after irradiation of UVA light. Conclusion. From the results reported above, it is concluded that the UVA-LT photocatalytic reaction have an antifungal effect on the denture surface Candida albicans, and so that could be applicable to the clinical use as a cleaning method.

Antimicrobial Active Substances from Entomopathogenic Fungi (Various Applications of Entomopathogenic Fungi)

  • Shin, Tae Young;Woo, Soo Dong;Kim, Jeong Jun
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.13-13
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    • 2016
  • Insects constitute the largest and most diverse group of animals in the world. They also serve as the hosts or nutrient sources for an immense assemblage of pathogens, parasites, and predators. More than 700 fungal species from 100 genera have adopted an entomopathogenic lifestyle. Although entomopathogenic fungi were studied as only biocontrol agents against a variety of pests in various countries, it has been recently focused their additional roles in nature. They are antagonists to/against plant pathogens, endophytes, and possibly even plant growth promoting agents. The potential antimicrobial effect against fungal plant pathogens by an isolate of entomopathogenic fungi including Beauveria bassiana, Lecanicillium spp., and Isaria fumosorosea have been reported since late 1990s, but wasn't reported pathogenicity of the isolate against pests. Later, a Canadian Lecanicillium sp. isolate and L. longisporium isolated from Vertalec$^{(R)}$ showed simultaneous control effect against both aphid and cucumber powder mildew. Therefore, the antimicrobial activities of 342 fungi isolates collected from various regions and conditions in Korea were evaluated against plant pathogenic fungus Botrytis cinerea using dual culture technique on agar plate. As a result, 186 isolates (54.4%) shown the antifungal activity against B. cinerea. The culture filtrates of selected fungi completely suppressed the growth of the microorganisms, indicating that suppression was due to the presence of antimicrobial substances in the culture filtrate. Mode of action of these fungi against insect involves the attachment of conidia to the insect cuticle, followed by germination, cuticle penetration, and internal dissemination throughout the insect. During infection process, secreted enzymes, proteinous toxins, and/or secondary metabolites secreted by entomopathogenic fungi can be used to overcome the host immune system, modify host behavior, and defend host resources. Recently, secondary metabolites isolated from entomopathogenic fungi have been reported as potential bioactive substances. Generally, most of bioactive substances produced by entomopathogenic fungi have reported low molecular weight (lower than 1,000 g/mol) as peptide and, in contrast the high molecular weight fungal bioactive substances are rare. Most substances based on entomopathogenic fungi were shown antimicrobial activity with narrow control ranges. In our study we analyzed the antimicrobial substances having antagonistic effects to B. cinerea. Antimicrobial substances in our fungal culture filtrates showed high thermostability, high stability to proteolytic enzymes, and hydrophilicity and their molecular weights were differed from substance. In conclusion, entomopathogenic fungi showed pathogenicity against insect pests and culture filtrate of the fungi also shown to antimicrobial activity. In the future, we can use the entomopathogenic fungi and its secondary metabolites to control both insect pest control and plant pathogenic fungi simultaneously.

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Ectomycorrhizal Development and Growth of Pinus thunbergii Seedlings Inoculated with Pisolithus tinctorius(KJ-1) in Copper Treated Soil (Cu를 처리(處理)한 토양(土壤)에서 Pisolithus tinctorius균(菌)을 접종(接種)한 곰솔유묘(幼苗)의 외생균근발달(外生菌根發達)과 생장(生長))

  • Oh, Kwang In;We, Kye Moon
    • Journal of Korean Society of Forest Science
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    • v.85 no.2
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    • pp.329-339
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    • 1996
  • This study is to investigate the effect of mycorrhizal development, growth, nutrient absorption of P. thunbergii seedlings inoculated with Pisolithus tinctorius(Pt. KJ-1) in relation to toxic materials in soil. The concentrations on copper solution applied to the soils were 0, 40, 120, 260, 430ppm. The results are summarized as follows : 1. The germination of P. thunbergii with Pt. increased greater than without ectomycorrhizal fungi. Mycorrhizal development showed a significant decreasing trend at high concentration, and tolerant Cu test with Pt. in agar plate media showed a decreasing trend at a high level. 2. P. thunbergii seedlings inoculated with Pt. showed that the shoot length was significantly promoted at 40, 120ppm copper levels, and that of noninoculated seedlings had the lowest effect in 430ppm copper level. 3. The outer shape of ectomycorrhizal root tips after inoculating Pt. on P. thunbergii seedlings appeared as a monopodial type, a fern-like type, and a cluster-like type at 0ppm, 40ppm levels, but only monopodial type came out at 260ppm, 430ppm copper levels. 4. Root length, no. of juvenile leaves, total length of juvenile leaves, total dry weight, no. of needles and total length of needles of P. thunbergii with Pt. increased greater than those of noninoculated seedling. Growth response of P. thunbergii seedling inoculated with Pt. increased significantly at 40ppm, 120ppm Cu levels. 5. As a result of analysis of growth medium, pH, Na, CEC increased at higher Cu level, and total nitrogen, organic matters, available phosphorous, K, Ca and Mg decreased at 40ppm, 120ppm Cu levels. As a result of an analysis about a copper within soils, it appeared having high Cu-concentration at 0ppm level of an inoculated Pt. and high Cu-concentration in noninoculated Pt. than in inoculated Pt. at higher level. 6. As a result of an analysis of shoot, N, P, and K-concentration were higher in noninoculated seedlings than in inoculated seedlings, and Cu-concentration was higher in inoculated seedlings than in noninoculated seedlings. The analysis of root resulted in a high N-concentration at 40ppm, 120ppm levels, in a high P-concentration in inoculated seedlings and in a high Cu-concentration in noninoculated seedlings. Cu-concentration was significantly higher at root than at shoot.

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