• Title/Summary/Keyword: Agar diffusion

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Antimicrobial Efficiency in the Fermented Slurry of Unpolished Rice (현미 발효 슬러리의 항균활성)

  • Choi, Hakjoon;Gwak, Gyeongja;Choi, Dabin;Park, Jaeyoung;Cheong, Hyeonsook
    • Microbiology and Biotechnology Letters
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    • v.43 no.4
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    • pp.307-313
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    • 2015
  • Unpolished rice (UR) is considered to be a healthy alternative to white rice when coping with chronic diseases. In the present study, the fermented slurry of unpolished rice (FSUR) was evaluated with respect to its antimicrobial activities and biochemical characteristics, including the quantities of sugar, total soluble sugar, organic acids, free amino acids, pH, and physiological activity. The antimicrobial efficiency of FSUR was assessed using the paper disc-agar diffusion method. FSUR exhibited strong antimicrobial activity against six pathogenic bacterial strains (Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, and Yersinia enterocolitica) and two fermentation strains (Gluconacetobacter intermedius and Lodderomyces elongisporus). The antimicrobial activity of FSUR was higher than the commercial antibiotics, carbenicillin ($50{\mu}g/ml$) and tetracycline ($50{\mu}g/ml$) against S. aureus, E. coli, L. monocytogenes, P. aeruginosa, S. typhimurium, Y. enterocolitica, and L. elongisporus. Also FSUR had a high antioxidant activity. The microorganisms were isolated from FSUR using tryptic soy broth and yeast extract-peptone-dextrose agar media. The isolated microorganisms were characterized using physiological and biochemical analyses as well as by 16S rRNA gene sequencing and phylogenic analysis. 16S rRNA gene sequence analysis showed that the isolated microorganisms had a high similarity to G. intermedius, Lactobacillus casei, Lactobacillus plantarum, and Acetobacter peroxydans.

Studies on Bovine Serum Protein Fractions (I, II) (혈청단백질분획(血淸蛋白質分劃)에 대한 연구(硏究) (I, II))

  • Rim, Bong Ho
    • Korean Journal of Veterinary Research
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    • v.5 no.1
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    • pp.1-16
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    • 1965
  • I. A Comparison of Sodium Sulfate Precipitation and Zone(Paper, Agar) Electrophoresis; Many kinds of techniques have been used for fractionating serum proteins. In the present study, using bovine serum, the fractions obtained with sodium sulfate were compared with those determined by zone electrophoresis. 1. Fibrinogen was precipitated with 4 to 10 percent of sodium sulfate. 2. ${\gamma}$-globulin required 10 to 16 percent of the salt for precipitation. 3. ${\beta}$-globulin began to precipitate at 12 percent sodium sulfate, and completed precipitation at approximately 26 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. 4. ${\alpha}$-globulin completed precipitation at 13 to 28 percent sodium sulfate in paper electrophoresis and at 22 percent in agar electrophoresis. 5. Albumin began to precipitate at 14 percent of the salt, and was free from the mixture of globulins approximately at 28 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. The results of comparing fractions by the two methods were as follows: 1. Euglobulin (15%) was equal to the sum of the most ${\gamma}$-globulin and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. 2. Pseudoglobulin I (15-17.5%) corresponded to the most ${\alpha}$-, ${\beta}$-globulins and a small quantity of albumin. 3. Pseudoglobulin II(18-22%) was a mixture of the ${\alpha}$-, ${\beta}$-globulins and albumin fraction. 4. Albumin (above 22%) contained the most albumin fraction separated by zone electrophoresis and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. As mentioned above the fractions obtained with sodium sulfate were a mixture of the various proportion of the fractions determined by zone electrophoresis. The solubility of serum fractions to sodium sulfate coincided with the mobility of those by zone electrophoresis. (By percent of sodium sulfate we mean gram of sodium sulfate contained in $100m{\ell}$ of solution). II. Immunological Studies on Serum Protein Fractions with Sodium Sulfate; In the previous report the fractions of bovine serum protein with sodium sulfate compared with those obtained by zone electrophoresis, and the findings were that the former contained various proportion components of the latter. In this study the author studied whether or not the fractions with sodium sulfate are simple component antigenically by immunoelectrophoresis and micro double diffusion test (Immuno-precipitation), using rabbit antiserum to bovine serum. In immunoelectrophoresis, normal bovine serum developed with rabbit antibovine serum showed about ten distinct precipitin arcs. The distribution of these arcs was as follows: 1 albumin, 2 ${\alpha}_1$-, 3 ${\alpha}_2$-, 2 ${\beta}_1$-, ${\beta}_2$-, and 1 ${\gamma}$-globulin (Fig. 7, 9). In micro double diffusion test, five to six precipitation bands could be seen between antigens and antibody, the order of the precipitation bands location is albumin, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-globulin from the side of antiserum well (Fig.19). Frequently the ${\alpha}$-, and ${\beta}$-precipitation bands were separated into two or three precipitation bands, which indicated that these globuline are not a pure component antigenically as shown in immuno-electrophoresis. In both Immunological methods, the two ${\alpha}$-, ${\beta}$-precipitin arcs and bands appeared clear and strong, indicating that the two globulins reacted as strong antigens. The precipitate reaction of ${\gamma}$-globulin was shown at 12 to 16 percent sodium sulfate; ${\beta}$-globulin at 12 to 20 percent; ${\alpha}$-globulin at 12 to 22 percent (immuno-electrophoresis), at 12 to 26 percent (Diffusion); and albumin at above 22 percent. Antigenically euglobulin contained ${\gamma}$-, ${\beta}$-, and ${\alpha}$-globulins, Pseudoglobulin I and Pseudoglobulin II were composed of ${\alpha}$-, and ${\beta}$-globulins, and albumin was a mixture of ${\alpha}$-globulin and albumin determined by zone electrophoresis. The results indicated that the fractions of serum protein obtained by either method were constituents of various proteins antigenically except ${\gamma}$-globulin and albumin by Zone electrophoresis.

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Screening of New Bioactive Materials from Microbial Extracts of Soil Microorganism (I) Antimicrobial Activity from 200 Sampled Using Microdilution Assay

  • Jung, Sang-Oun;Kim, Joon;Chang, Il-Moo;Ryu, Jae-Chun
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.278-285
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    • 1998
  • The microdilution assay recommended by NCCLS (National Committee for Clinical Laboratory Standards) is one of the standardized methods of antibiotic susceptibility test. This method has been widely used clinically to obtain MIC values of antibiotics on pathogenic microorganisms. It is more convenient, rapid and simple to test many samples than other test methods such as agar diffusion assay and broth macrodilution assay. The screening of antimicrobial agents from microbial extracts is too laborious in its process. Therefore, a number of screening methods having more simple procedure have been developed. In our laboratory, we applied microdilution assay for screening the antimicrobial agents. This assay showed dose-response results and was more sensitive than disc diffusion assay in our system. We tested 200 samples of microbial extracts originated from 100 microbial strains and selected several samples as potential candidates. In this report, we show that the microdilution assay is more convenient method in screeing of antibiotic susceptibility than those previously reported.

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Bacteriocins: Assay, Biochemistry, and Mode of Action

  • Paik, Hyun-Dong
    • Preventive Nutrition and Food Science
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    • v.1 no.2
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    • pp.269-277
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    • 1996
  • Bacteriocins are proteins produced by a heterogeneous group of bacteria that have a bactericidal effect on closely related organisms. Recently, bacteriocins from lactic acid bacteria and other food-related organisms have been the subject of much research because of their potential as food biopreservatives. Various modifications of agar plate diffusion assays are the most widely used methods even though the limitations of such assays are generally recognized. The ability to obtain a concentrated crude preparation on bacteriocin by optimizing production parameters greatly simplifies recovery of bacteriocin on subsequent purification steps. Some studies performed to optimize bacteriocins have been purified to homogeneity, and the amino acid sequences of many of these purified bacteriocins have been determined. Obtaining characterization data on purified bacteriocin will minimize the risk of overlapping of research and confusion on identification of these compounds. Several me-chanisms leading to cell death have been hypothesized. These include depletion of the proton motive force(PMF) across the cell membrane: RNase and/or DNase activity within the sensitive cell; and pore formation and lysis of sensitive cells at the cell membrane.

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Anti-microbial Activities Against Oral Microbes and Growth-Inhibitory Effect on Oral Tumor Cell of Extracts of Perilla and Mugwort (들깨잎과 쑥 추출물의 구강병 원인균에 대한 항균 및 KB 세포 증식 억제효과)

  • Jo, Min-Jung;Min, Kyung-Jin
    • Journal of Environmental Health Sciences
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    • v.33 no.2 s.95
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    • pp.115-122
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    • 2007
  • Methanol extracts of Perilla and Mugwort were stepwise extracted with hexane, chloroform, ethyl acetate, butanol and water. Anti-microbial activities and inhibitory effect on growth of oral tumor cell of each extract were investigated. Each extracts of Perilla and Mugwort were investigated to anti-microbial effects on oral microbes by means of agar diffusion method and MIC. These results suggest that the hexane extracts of Perilla and Mugwort have antimicrobial activities against S. mutans and potent inhibitory Effect to KB cell growth.

Selection of Human-Originated Lactobacillus acidophilus For Production of Probiotics

  • Kim, Wang-June;Hong, Seok-San;Cha, Seong-Kwan
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.151-154
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    • 1994
  • Lactobacillus acidophilus KFRI 233, a strain isolated from human, was selected as a candidate for probiotics due to its excellent growth in MRS broth where no special anaerobic condition is required. Both simultaneous and deferred agar diffusion assays exhibited Lb. acidophilus KFRI 233 to possess an antagonistic effect against Clostridium perfringens. Its antagonistic effect was pH dependent Associative culture of KFRI 233 and Cl. perfringens in broth resulted in maximum 94.04% inhibition of Cl. perfringens. $\beta$-Galactosidase activity of KFRI 233 was higher than other tested strains that are sold as commercial probiotics. Survival of KFRI 233 in pasteurized skim milk (4$^{\circ}C$) and Sherbet mix (-15$^{\circ}C$) after 7 days of storage were 71.9 and 105.5%, respectively.

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Actinoplanes teichomyceticus의 변이주에 의한 Teicoplanin 발효생산

  • Kim, Seong-Geon;An, Hyeon-Jin;Kim, Jae-Yeong;No, Yong-Taek
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.299-302
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    • 2000
  • Actinoplanes teichomyceticus ATCC 31121 was mutated with UV to obtain a superior mutant strain with increasing level of teicoplanin production. In this investigation lethal curve was obtained and the optimal condition to induce mutagenesis was determined and to isolate the desirable mutant strain. It was also confirmed that teicoplanin activities by agar diffusion method to be compared to the mother strain. One mutant strain, T991014-1 that had the highest teicoplanin productivity was finally selected for further investigation including fermentation pattern. The mutant was characterized by the various tests such as amylase activity, protease activity, antibiotic resistance, autotoxicity, and productivity.

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Characterization of Bacteriocin produced Lactobacillus bulgaricus acting on bovine mastitis pathogens

  • Kim, Hyun-Jin;Kim, Ji-Hyun;Kim, Sung-Koo
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.740-744
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    • 2003
  • The antimicrobial substance produced by Lactobacillus bulgaricus was inactivated by pretense, which confirmed it as a bacteriocin and referred to 'bulgaricin HJ'. The bulgaricin HJ showed the inhibitory activity against mastitis pathogens, gram-positive and gram-negative bacteria. The optimal conditions for the production of bulgaricin HJ were at the temperature of $30^{\circ}C$ and 10 h after cultivation of L. bulgaricus. Staph. and Strep. agalactiae, common bovine mastitis pathogens, were treated with bulgaricin HJ by the agar well diffusion method and showed antimirobial activities to the bovine mastitis pathogens. The activity of the bulgaricin HJ was maintained at pH 6-7 and $100^{\circ}C$ for 60 min against the mastitis pathogens. The bulgaricin HJ was determined as class IV bacteriocin by various enzyme treatments. Colony forming units analysis with indicator strains by the treatments of bulgaricin HJ indicates that the mode of bacteriocin action was bactericidal rather than bacteriostatic.

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Antibiotics susceptibility and biochemical characterization of Escherichia coli associated with diarrhea from preweaning piglets (포유자돈 소장에서 분리된 대장균의 생화학 성상과 항생제 감수성 결과)

  • Ham, Hee-jin;Min, Kyoung-sub;Chae, Chan-hee
    • Korean Journal of Veterinary Research
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    • v.37 no.4
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    • pp.773-777
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    • 1997
  • A retrospective study of 132 cases of natural Escherichia coli infection in preweaning piglets with diarrhea submitted to the Department of Veterinary Pathology in Seoul National University from 1995 to 1996 was performed to determine the biochemical characteristics and antibiotics susceptibility. Most field isolates were lysine decarboxylase-positive (99.2%), phenylalanine decarboxylase-negative (100%), and fermented sorbitol (91.7%). Antibiotic susceptibilities will be determined by agar diffusion method. A large percentage of isolates were resistant to many antibiotics in common usage. Most isolates were susceptible to colistin (98.8%), gentamycin (64.3%), amikacin (100%), and ceftiour sodium (64.2%), whereas most isolates were notably resistant to ampicillin (86.9%), neomycin (66.7%), streptomycin (84.5%), tetracycline (98.8%), penicillin (98.8%), and amoxacillin (58.3%).

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Effects of Edible Herbs on the Growth of In Vitro Intestinal Microorganisms (산채류가 장내세균의 In Vitro 생육에 미치는 영향)

  • 한복진
    • Journal of Nutrition and Health
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    • v.27 no.7
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    • pp.717-728
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    • 1994
  • This study was aimed to screen edible herbs which control the composition of intestinal microflora. With in vitro experiments, we screened the water or ethanol extracts of about 60 edible herbs and wild plants in terms of the inhibition activity on the growth of the harmful Clostridium perfringens and growth promoting activity for the beneficial Bifidobacteria. The water extracts of mugwort and small water dropwort inhibited the growth of Cl.perfringens both in agar diffusion method and broth culture. On the other hand, the water extracts of petasites, mugwort, yellow day-lily and bitter cress have shown the promotion effect on the growth of Bifidobacterium longum. In the culture test using human feces as starter, the extracts of the above selected herbs increased the population of Bifidobacteria and Lactobacillus while they reduced the numbers of Cl.perfringens and E.coli.

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