• Title/Summary/Keyword: Agar gel

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Breeding of Aspergillus oryzae for the Alkaline Pretense Overproducing Strain. (재조합 Alkaline Protease를 대량 생산하는 Aspergillus oryzae 균주개발)

  • 이병로;유기원;최원균;최동성;임한진;성창근
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.450-455
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    • 1998
  • Aspergillus oryzae M-2-3 strain (argB$\^$-/) was transformed with pTAalp plasmid which was constructed for expression of the alkaline pretense gene, alpA, and 16 transformants were selected on arginine minus medium. When these transformants were tested for productivity of alkaline proteases using agar plate containing skim milk, the halo was observed around each colony of transformants, but not observed around the host strain in this condition. Southern analysis showed that the pTAalp plasmid having alpA gene was integrated into the chromosome of the host strain. The highest level of alkaline protease production was obtained in the culture filtrate of the transformant No. 14, which was estimated to 80-90% of total secreted proteins, and the enzyme activity was 64-450 times higher than those of host strain and industrial strain. Total nitrogen content and the digestion rate in soybean Koji extracts were also increased to 1.5 times in Aspergillus oryzae transformant No. 14.

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Analysis and Quantification of Ammonia-Oxidizing Bacteria Community with amoA Gene in Sewage Treatment Plants

  • Hong, Sun Hwa;Jeong, Hyun Duck;Jung, Bongjin;Lee, Eun Young
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1193-1201
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    • 2012
  • The analysis and quantification of ammonia-oxidizing bacteria (AOB) is crucial, as they initiate the biological removal of ammonia-nitrogen from sewage. Previous methods for analyzing the microbial community structure, which involve the plating of samples or culture media over agar plates, have been inadequate because many microorganisms found in a sewage plant are unculturable. In this study, to exclusively detect AOB, the analysis was carried out via denaturing gradient gel electrophoresis using a primer specific to the amoA gene, which is one of the functional genes known as ammonia monooxygenase. An AOB consortium (S1 sample) that could oxidize an unprecedented 100% of ammonia in 24 h was obtained from sewage sludge. In addition, real-time PCR was used to quantify the AOB. Results of the microbial community analysis in terms of carbon utilization ability of samples showed that the aeration tank water sample (S2), influent water sample (S3), and effluent water sample (S4) used all the 31 substrates considered, whereas the AOB consortium (S1) used only Tween 80, D-galacturonic acid, itaconic acid, D-malic acid, and $_L$-serine after 192 h. The largest concentration of AOB was detected in S1 ($7.6{\times}10^6copies/{\mu}l$), followed by S2 ($3.2{\times}10^6copies/{\mu}l$), S4 ($2.8{\times}10^6copies/{\mu}l$), and S3 ($2.4{\times}10^6copies/{\mu}l$).

Serological survey of diseases to poultry and swine in slaughtered ostriches (도축 타조에서 닭 및 돼지 질병에 대한 혈청학적 조사)

  • Kim Soon-Tae;Park In-Hwa;Kim Young-Hoan;Cho Kwang-Hyun;Oh Kyu-Shil;Son Jae-Kweon;Jyeong Jong-Sik
    • Korean Journal of Veterinary Service
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    • v.27 no.3
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    • pp.281-288
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    • 2004
  • As all other intensively farmed domestic species, most mortality in ostriches is closely to rearing conditions. While ostriches is also highly sensitive to stress, species-specific infectious disease play only a minor role. But investigation of ostrich's disease is not peformed almost in Korea. The study was performed to investigate the titers of antibody for Newcastle disease(ND), Infectious bronchitis(IB), Egg drop syndrome '76(EDS), Avian influenza(AI), salmonellosis, Mycoplasma gallisepticum infection(MG), Mycoplasma synoviae infection(MS), Infectious bursal disease(IBD), Brucellosis, Toxoplasmosis, Japanese encephalitis(JE), Porcine parvovirus infection, Encephalomyocarditis and Porcine reproductive respiratory syndrome (PRRS). The results obtained in the 62 ostrich sera slaughtered in Gyeongbuk province were summarized as follows: The average of antibody positive rates to ND, IB, EDS, AI(H9Nl), JE, Porcine parvovirus infection and Encephalomyocarditis by HI test were $75.8\%,\;100\%,\;0\%,\;0\%,\;51.6\%,\;50\%\;and\;56.5\%$ respectively. The antibody positive rates to salmonellosis, MG, MS by plate agglutination test were $12.9\%,\;25.8\%,\;and\;0\%$ respectively. Antibodies to disease agent such as IBD and AI by agar gel precipitation(AGP) test, Brucellosis by tube agglutination, toxoplasmosis by latex agglutination test and PRRS by IFA were all negative.

Characteristics of Cytophaga columnaris isolated from rainbow trout (Oncorhynchus mykiss), goldfish(Carassius auratus), and ayu(Plecoglossus altivels) in Korea (국내에서 양식되는 무지개송어(Oncorhynchus mykiss), 금붕어(Carassius auratus), 은어(Plecoglossus altivelis) 로부터 분리된 Cytophaga columnaris의 특성)

  • Park, Se-chang;Heo, Gang-joon
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.371-378
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    • 1998
  • Five strains of gram-negative and yellow-pigmented bacteria were recently isolated from diseased freshwater fishes in Korea. All isolates were confirmed as a known fish pathogen of columnaris disease, Cytophaga columnaris based on their colonial and cellular morphology, and on physiological, biochemical and antigenic characteristics. Although the isolates were from different fish species, their characteristics of them were very similar to those of the reference strains of C columnaris (NCMB $2248^T$ and EK 28). Also, profiles of OMPs of Korean isolates were similar to those of the reference strain, C columnaris NCMB $2248^T$ when analyzed by SDS-PAGE.

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Isolation and Structure Identification of Phytotoxins from Lasiodiplodia theobrorme, the Cause of JAVA Black Rot of Sweet Potato (고구마 검은썩음병균(Lasiodiplodia theobromae)으로부터 식물독소의 분리 및 구조)

  • Lim, Chi-Hwan
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.118-123
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    • 1998
  • Lasiodiplodia theobrorme is a pathogen of Java black rot on sweet potato. This fungus infects the tuberous root during storage after harvest. Invasion of the fungus results in the expansion of necrosis sites into the tuberous roots. The resultant necrotic symptom of the tissue is also induced by application of acetone extract of the fungus growing on potato sucrose agar (PSA) culture. The active principles to induce the necrosis are purified from the acetone extract as follows. After evaporation of hexane-benzene-EtOAc (1:2:1, v/v/v) the extract was fractioned on silica gel column, using a solvent gradient system from n-hexane to EtOAc and then to MeOH. The active fractions were purified with HPLC on Nucleosil 50-5 column by eluting n-hexane to EtOAc. Their structures are established by using spectroscopic techniques and synthesis to 4-hydroxymellein and mellein, respectively. Application of small amount of these compounds induce expansion of the necrotic symptom into the tissue and accumulated ipomeamarone. Conclusively, these compounds acted as phytotoxins (inducing necrosis) and as elicitors (eliciting the phytoalexin).

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Hand Hygiene Effects Measured by Hand Culture in Intensive Care Unit (배양검사를 통해 확인한 중환자실 직원의 손씻기 효과)

  • Jeong, Jae-Sim;Choi, Jeong-Hwa;Lee, Soon-Haeng;Kim, Yang-Soo
    • Journal of Korean Biological Nursing Science
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    • v.5 no.2
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    • pp.21-30
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    • 2003
  • The effect of hand hygiene was measured by hand culture before and after hand hygiene for 86 nurses, doctors, and nurses aide/housekeepers in Surgical Intensive Care Unit. The subjects were asked to press their dominant hand in hand-shaped Mannitol salt agar immediately after patient contact and then washed their hand by preferred hand hygiene agents [soap and water, waterless alcohol gel, or 4% chlorhexidine gluconate detergent (CHG)], and cultured one hand again Amount of isolated microorganism was calculated by counting the number of divided areas ($1{\times}1cm$) which is culture positive in hand culture plate. The amount of microorganisms were significantly reduced from 58.1(${\pm}38.59$) to 27.4(${\pm}30.4$) cells after hand hygiene. The staff nurse's hand hygiene was more effective compared to medical doctors and nurses aide/housekeepers. Methicillin-resistant Staphylococcus aureus(MRSA) was isolated in 41(47.1%) subjects ; but only removed 100% in 28(32.2%) subjects. When the amount of hand microorganisms was compared by subject's preferred hand hygiene agents, it was decreased in order of 4% CHG, waterless alcohol solution, soap and water, and water. The hand hygiene practice was inadequate to reduce hand microorganisms and significantly different by occupations. Further research and development of hand hygiene improvement program which emphasize the quality of hand hygiene is recommended.

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Two New PVY Strains Isolated from Tobacco Plant in Korea (한국 잎담배에서 분리된 PVY계통)

  • Park E. K.;Kim J. J.;Boo K. S.
    • Korean journal of applied entomology
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    • v.23 no.4 s.61
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    • pp.209-214
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    • 1984
  • Since 1980, burley tobacco plants grown mainly in the western hat of the Korea. have shown two new types of disease symptom. Both symptoms were found to be caused by two different PVY strains : the vein banding type by a PVY strain designated as PVY-VB and necrosis on leaf veins by a PVY strain designated as PVY-VN, Identification of the PVY strains was based on host range test. aphid( Myzus Persicae) transmission test, physical properties, serology, and observation of virus particle morphology. The virus particles were measured to be about 730 nm without any difference in shape or dimensions between the two strains. Both strains also gave a positive reaction to the PVY antiserum in SDS-agar gel double diffusion test. These strains, however, gave a negative reaction to the tobacco etch virus and tobacco vein mottling virus antisera.

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Isolation and Evaluation of Protective Effect against Fusarium Wilt of Sesame Plants of Antibiotic Substance from Bacillus polymyxa KB-8

  • Hyun, Jae-Wook;Kim, Young-Hoon;Lee, Yong-Se;Park, Won-Mok
    • The Plant Pathology Journal
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    • v.15 no.3
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    • pp.152-157
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    • 1999
  • An antibiotic compound was isolated from the culture of an antagonist against Fusarium oxysporum f. sp. sesami, Bacillus polymyxa strain KB-8, and tested for the control of Fusarium wilt of sesame in greenhouse conditions. Optimum conditions for culturing the antagonist to obtain the maximum antibiotic activity were determined using different culture media, initial medium acidity, and incubation periods for which yeast -malt extract agar with the initial acidity of pH 5 and over 13 days culture were best. Antibiotic substances extracted by methanol had 2 main fractions, KB-8A and KB-8B, in thin layer chromatography (OLC) with Rf values of 0.35 and 0.67 in a solvent system of chloroform : methanol = 7 : 3. The fraction KB-8A wa purified further by XAD-2, silica gel and Sephadex LH-20 column chromatography, and crystalization. Its minimum inhibitory concentrations (MICs) were $12.8\mu\textrm{g}$/ml for F. oxysporum and Alternaria mali, $6.4\mu\textrm{g}$/ml for Colletotrichum gloeosporioides and Rhizoctonia solani, and $3.2\mu\textrm{g}$/ml for Phytophthora capsici. Soil drenching of antibiotic KB-8A in the concentrations of $13.0\mu\textrm{g}$/ml and $26.0\mu\textrm{g}$/ml effectively inhibited the Fusarium wilt of sesame in a greenhouse test, which appeared to be comparable to the fungicide benlate of $6.5\mu\textrm{g}$ a. i./ml.

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Candida non albicans with a High Amphotericin B Resistance Pattern Causing Candidemia among Cancer Patients

  • Kalantar, Enayatollah;Assadi, Mojan;Pormazaheri, Helen;Hatami, Shiva;Barari, Maryam Agha;Asgari, Esfandiar;Mahmoudi, Elaheh;Kabir, Kourosh;Marashi, Seyed Mahmoud Amin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10933-10935
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    • 2015
  • Background: Many scientists have reported Candida species to be of great concern because of the high frequency that they colonize and infect human hosts, particularly cancer patients. Moreover, in the last decades Candida species have developed resistance to many antifungal agents. Based on this, we aimed to identify and determine the prevalence of Candida spp from blood culture bottles among cancer patients and their antifungal resistance pattern. Materials and Methods: From the blood culture bottles isolation and identification of the Candida spp were performed by conventional microbiological techniques. The in vitro antibiotic resistance pattern of the isolates was determined by CLSI guidelines. Genomic DNA was isolated and amplified. Each gene was separated by agar gel electrophoresis. Results: Identification of Candida spp was based on the presence of yeast cells in direct examination, culture and DNA extraction. Of the 68 blood samples collected during the study period (April 2013 to October 2013), five (7.35%) were positive for the presence of Candida spp, 2 (40%) of which were identified as Candida albicans and 3 (60%) were Candida non-albicans. Conclusions: High resistance to amphotricin B was observed among all the Candida non-albicans isolates. Regular investigations into antifungal resistance will help us to get an updated knowledge about their antibiotic resistance pattern which may help the physician in selecting the antibiotics for empirical therapy.

Proteomic Analysis of Proteins Increased or Reduced by Ethanol of Lactobacillus plantarum ST4 Isolated from Makgeolli, Traditional Korean Rice Wine

  • Lee, Seung-Gyu;Lee, Kang-Wook;Park, Tae-Heung;Park, Ji-Yeong;Han, Nam-Soo;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.516-525
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    • 2012
  • LAB were isolated from makgeolli locally produced around Jinju, Gyeongnam, S. Korea during spring of 2011. Randomly selected 11 isolates from MRS agar plates were identified first by API CHL 50 kits and then 16S rRNA gene sequencing. All 11 isolates were identified as Lactobacillus plantarum. Among them, ST4 grew in MRS broth with ethanol up to 10%, showing the highest alcohol resistance. L. plantarum ST4 was moderately resistant against acid and bile salts. When cellular proteins of L. plantarum ST4 under ethanol stress were analyzed by two-dimensional gel electrophoresis (2DE), the intensities of 6 spots increased, whereas 22 spots decreased at least 2-fold. Those 28 spots were identified by peptide mass fingerprinting (PMF). FusA2 (elongation factor G) increased 18.8-fold (6% ethanol) compared with control. Other proteins were AtpD (ATP synthase subunit beta), DnaK, GroEL, Tuf (elongation factor Tu), and Npr2 (NADH peroxidase), respectively. Among the 22 proteins decreased in intensities, lactate dehydrogenases (LdhD and LdhL1) were included.