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Optimum Cultivation Conditions for Mass Production of an Antagonistic Bacterium Bacillus subtilis BD0310 for Development of a Microbial Agent Controlling Gray Blight of Tea Plants (차나무 겹둥근무늬병 방제용 미생물제제 개발을 위한 길항세균 Bacillus subtilis BD0310의 대량배양 최적조건)

  • Kim Gyoung-Hee;Oh Soon-Ok;Hur Jae-Seoun;Yum Kue-Jin;Koh Young-Jin
    • Research in Plant Disease
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    • v.12 no.2
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    • pp.85-90
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    • 2006
  • Bacillus subtilis BD0310 isolated from tea leaves was used for the development of a biofungicide against Pestalotiopsis longiseta causing gray blight of tea plants. The optimum growth conditions were investigated for the mass cultivation of the microbial agent. The optimum temperature and cultivation time were determined as $12{\sim}24$ hours at $30^{\circ}C$ and the optimum initial pH was pH 7.0 in nutrient broth. Among the tested carbon sources of fructose, galactose, glucose, glycerol, inositol, lactose, maltose, sorbitol and starch, maltose and inositol were found to highly increase antifungal activity of the microbial agent against P. longiseta. Yeast extract and tryptone apparently increased antifungal activity of the microbial agent among the tested nitrogen sources of casein, tryptone, malt extract, yeast extract and $(NH_4)_2SO_4$. The results will make a contribution to mass production of the antagonistic bacterium Bacillus subtilis BD0310 for development of a microbial agent controlling gray blight of tea plants.

Oropharyngeal Carriage and Antimicrobial Resistance of S. pneumoniae in Children of Seoul (서울 지역 소아의 구인강에서 폐구균 보균율과 항균제 내성)

  • Kim, Young Kee;Lee, Chang Kyu
    • Pediatric Infection and Vaccine
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    • v.4 no.2
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    • pp.218-224
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    • 1997
  • Purpose: The antimicrobial resistance of S. pneumoniae has encountered with increasing frequency from around the world. In our country, penicillin resistant strains of S. penumococci are rapidly increasing. It has been known that colonized pneumococci in upper respiratory tract cause sinisitis, otitis media, meningitis and pneumonia. We tried to reveal the colonization rate of pneumonocci in upper respiratory tract, their antimicrobial resistance and DNA fingerprinting pattern in normal children. Methods: We got specimens from 117 children of day-care center in Seoul through oropharyngeal swab. After incubation on BAP, optochin test and slide latex agglutination test were used for identification. Antimicobial susceptibility test to penicillin, vancomycin, erythromycin and TMP-SMZ was done with disk diffusion method. Penicillin MIC was gotten through the broth microdilution method. Genotyping of 45 pneumococci was done by rep-PCR using REP1R-Dt and REP2-Dt primer. Results: The carriage rate of pneumococci in the day-care center children was 38%(45/117). The resistance of penicillin, erhthromycin, TMP/SMZ, vancomycin by the disk diffusion method are 89%, 91%, 64% and 0%, respectively. 64% of the isolates showed multiple resistance. 7 types of DNA fingerprinting were gotten and 78% of isolates belonged to three types. Conclusion: We found that the antimicrobial resistance of children attending the day-care center in Seoul was much higher than expected. We assumed that this might be due to their easy and frequent exposure to antimicrobial agents and crowded day-care center environment.

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Production and Characterization of Alkaline Protease of Micrococcus sp. PS-1 Isolated from Seawater (해수에서 분리한 Micrococcus sp. PS-1이 생산하는 단백질 분해효소의 생산과 효소학적 특성)

  • Jin, Young-Rang;Yu, Sun-Nyoung;Kim, Kwang-Youn;Kim, Sang-Hun;Park, Seul-Ki;Kim, Hyeun-Kyeung;Lee, Yong-Seok;Choi, Yong-Lark;Ji, Jae Hoon;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.23 no.2
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    • pp.273-281
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    • 2013
  • The purpose of this research was to investigate the production and characterization of alkaline protease from Micrococcus sp. PS-1 newly isolated from seawater. Micrococcus sp. PS-1 was grown in Luria-Bertani (LB) medium. Its optimal temperature and pH for growth were $30^{\circ}C$ and 7.0, respectively. The effect of nitrogen sources was investigated on optimal enzyme production. A high level of alkaline protease production occurred in LB broth containing 2% skimmed milk. The protease was purified in a 3-step procedure involving ultrafiltration, acetone precipitation, and dialysis. The procedure yielded a 16.43-purification fold, with a yield of 54.25%. SDS-PAGE showed that the enzyme had molecular weights of 35.0 and 37.5 kDa. Its maximum protease activity was exhibited at pH 9.0 and $37^{\circ}C$, and its activity was stable at pH 8.0-11.0 and $25-37^{\circ}C$. The protease activity was strongly inhibited by PMSF, EDTA, and EGTA. Taken together, the results demonstrate that the protease enzyme from Micrococcus sp. PS-1 probably belongs to a subclass of alkaline metallo-serine proteases.

A Comparative Study between Microbial Fermentation and Non-Fermentation on Biological Activities of Medicinal Plants, with Emphasis on Enteric Methane Reduction (천연 약용식물의 미생물 발효를 통한 장내 메탄 생성 억제 효과 비교 연구)

  • Lee, A-Leum;Park, Hae-Ryoung;Kim, Mi-So;Cho, Sangbuem;Choi, Nag-Jin
    • Korean Journal of Organic Agriculture
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    • v.22 no.4
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    • pp.801-813
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    • 2014
  • A study was conducted to improve the biological activity of two medicinal plants, Eucommia ulmoides Oliv. and Glycyrrhiza uralensis, by fermentation. The biological activity was assessed by determining antibacterial, antioxidant and antimethanogenic properties. Fermentation was achieved by adding the plant materials in MRS broth at 10% (w/v) and different starter cultures at 1% (v/v). Condition for fermentation were incubation temperature of $30^{\circ}C$ and agitation at 150 rpm for 48 h. Six starter cultures, Weissella confusa NJ28 (Genbank accession number KJ914897), Weissella cibaria NJ33 (Genbank accession number KJ914898), Lactobacillus curvatus NJ40 (Genbank accession number KJ914899), Lactobacillus brevis NJ42 (Genbank accession number KJ914900), Lactobacillus plantarum NJ45 (Genbank accession number KJ914901) and Lactobacillus sakei NJ48 (Genbank accession number KJ914902) were used. Antibacterial activity was observed in L. curvatus NJ40 and L. plantarum NJ45 only as opposed to other treatments, including the non-fermented groups, which showed no antibacterial activity. Both plants showed antioxidant activity, although E. ulmoides Oliv. had lower activity than G. uralensis. However, fermentation by all strains significantly improved (p<0.05), antioxidant activity in both plants compared to non-fermented treatment. Six treatments were based on antibacterial activity results, selected for in vitro rumen fermentation; 1) non-fermented E. ulmoides, 2) fermented E. ulmoides NJ40, 3) fermented E. ulmoides NJ45, 4) non-fermented G. uralensis, 5) fermented G. uralensis NJ40, 6) fermented G. uralensis NJ45. A negative control was also added, making a total of 7 treatments for the in vitro experiment. Medicinal plant-based treatments significantly improved (p<0.05) total volatile fatty acid (VFA) concentration. Significant methane reduction per mol of VFA were observed in G. uralensis (p<0.05). Based on the present study, fermentation improves the biological activity of E. ulmoides Oliv. and G. uralensis. Fermented G. uralensis could also be applied as an enteric methane mitigating agent in ruminant animals.

Biological Evaluation of Nargenicin and Its Derivatives as Antimicrobial Anti-inflammatory Agents (토양 균주 발효 추출물 Nargenicin 및 그 유도체의 항생제 대체 효과능 평가)

  • Cho, Seung-Sik;Hong, Joon-Hee;Chae, Jung-Il;Shim, Jung-Hyun;Na, Chong-Sam;Yoo, Jin-Cheol
    • Korean Journal of Organic Agriculture
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    • v.22 no.3
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    • pp.469-481
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    • 2014
  • IIn vitro antimicrobial and anti-inflammatory activities of nargenicin and its derivatives were investigated. Nargenicin, an unusual macrolide antibiotic with potent anti-MRSA (methicilin-resistant Staphylococcus aureus) activity, was purified from the culture broth of Nocardia sp. CS682. And variety of novel nargenicin derivatives was synthesized from nargenicin. Two compounds (4 and 5) exhibit a broad spectrum of antimicrobial activities against infectious bacteria. The antimicrobial activity of derivatives against fifteen organisms was assessed using the minimum inhibitory concentration (MIC). The MIC values were in the ranges of $0.15{\sim}80{\mu}g/mL$ (w/v) for compound 1 and 2, $5{\sim}80{\mu}g/mL$ (w/v) for compound 3, $1.25{\sim}40{\mu}g/mL$ (w/v) for compound 4, and $1.25{\sim}80{\mu}g/mL$ (w/v) for compound 5, depending on the pathogens studied. In vitro, we investigated cytotoxicity and inhibition of nitric oxide (NO) production of synthesized compounds 1-5 in Raw 264.7 cells. LPS-induced nitric oxide releases were significantly blocked by compound 3, 4 and 5 in a dose-dependent manner. At high concentrations ($5{\mu}g/mL$) compound 5 inhibited the NO production by 95%. Compound 4 inhibited the release of NO in LPS-activated Raw 264.7 cells by 75% at the concentration of $10{\mu}g/mL$. Compound 3 inhibited the release of NO in LPS-activated Raw 264.7 cells by 65% at the concentration of $100{\mu}g/mL$. On the other hand, nargenicin, compound 1 and 2 did not inhibit NO production. These results demonstrated that compound 4 and 5 displayed antimicrobial activity and blocked LPS-induced pro-inflammatory mediators such as NO in macrophages, which might be responsible for its therapeutic application.

Bacteriological Study of Listeria sp. Isolated from Seawater and Sea Food (해수와 해산물로부터 Listeria 속의 분리와 세균학적 조사)

  • 강치희;이만효;황용일
    • Journal of Life Science
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    • v.13 no.4
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    • pp.390-399
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    • 2003
  • Four species of the genus of Listeria were isolated from seawater and sea food in Kyungnam province, South Korea. These isolated strains were classified into Listeria sp. from different samples by appropriate cultivation conditions and biochemical tests including serological test. In a day enrichment cultivation, the following strains were found out of 100 samples: L. innocua (35%), L. ivanovii (4%), L. monocytogenes (4%), and L. welshimeri (1%). For seven days enrichment culture, L. innocua (38%), L. ivanoii (5%), L. monocytogenes (7%), and L. welshimeri (1%) were isolated. From these results, Listeria species were more efficiently isolated in seven day enrichment broth than in one day enrichment. However, these isolated Listeria species were less grown in the selective medium than in the enrichment medium. Isolation rates of Listeria species showed differency for each sample and Listeria species were more abundantly isolated in shrimps (80%) and crayfishes (80%) than little neck clams (50%), seawater (25%) and mussels (20%). From the results of serological classes for the seven L. monocytogenes, two strains were defined as type I and the other five strains as type IV.

Studies on the Production of Lysine by Fermentation Process (2) -Lysine Production by Auxotrophs- (발효에 의한 라이신(L-Lysine) 생산에 관한 연구 (2) -영양요구성 변이주에 의한 Lysine 생산-)

  • Min, Tae-Ick;Kim, Hang-Mook;Kwon, Tai-Wan
    • Korean Journal of Food Science and Technology
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    • v.4 no.2
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    • pp.123-133
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    • 1972
  • Over 90 of lysine producing auxotrophs were obtained from Corynebacterium sp. S-27-12, Brevibacterium flavum ATCC 15168 and Micrococcus glutamicus ATCC 13032 by UV light, $Co^{60}$ irradiation and N-methyl-N'-nitro-N-nitrosoguanidine treatment. One of the mutant, Brev. flavum U46-N59, was identified as a leucine auxotroph and accumulated lysine during flask (500 ml) cultivation (180 strokes/min.) up to 21.6 mg per ml of broth at pH 7.5 and $28^{\circ}C$ after 4 days. The medium consisted of glucose, 100; urea, 10; corn steep liquor, 40; $KH_2PO_4,\;2;\;K_2HPO_4,\;0.5;\; MgSO_4.\;7H_2O,\;0.4;\;antifoam\;S-57,\;1g;\;Fe_2(SO_4)_3.XH-2O,\;10;\; MnCl_2,\;4H_2O,\;10mg;\;biotin,\;30;\;thiamine-HCl,\;100{\mu}g$in 1l of distilled water, and 40 U/ml of penicillin was added after 36 hrs fermentation.

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Microbiological and Immunological Investigation on the Bacteroides gingivalis in Rapidly Progressive and Adult Periodontitis in Korean (한국인 급성진행성 및 성인성 치주염의 원인균인 Bacteroides gingivalis에 대한 미생물 및 면역학적 연구)

  • Chung, Chong-Pyoung;Lee, Jong-Heun;Chung, Hyun-Ju
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.309-321
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    • 1987
  • For the investigation of microbiological and immunological specificity of Bacteroides gingivalis, Bacteroides gingivalis were isolated, enumerated and characterized from 13 Korean rapidly progressive periodontitis and 7 healthy control by anaerobic culture technique. The total proportion of black-pigmented Bacteroides from Korean R.P.P. patients and healthy control were 8.78% and 0.92%, respectively, among total isolated black-pigmented Bacteroides. In antibiotic susceptibility test, Bacteroides gingivalis isolated from R.P.P. patients were sensitive to Ampicillin and Tetracycline, and resistant to Gentamicin and Erythromycin in disc diffusion method. In antibiotic broth dilution method, the minimum inhibitory concentration(MIC) to Bacteroides gingivalis was 2 unit/ml of Penicillin and $0.25{\sim}1{\mu}g/ml$ of Tetracycline, respectively. The concentration of serum IgG in rapidly progressive periodontitis patients were sigificantly higher than that of healthy control, and concentration of diluted gingival crevicular IgG has not any significant differences between two groups. Serum and gingival crevicular IgG antibody to Bacteroides gingivalis were significantly higher titer in rapidly progressive periodontitis patients to compare with healthy control. The lipopolysaccharide profiles of 2 Korean B. gingivalis in silver stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis were similar to type strains of B. gingivalis and typical LPS band were appeared around the 24-Kd molecular weight. Immunodiffusion test and immunoelectrophoresis of the L.P.S. extracted from 2 Korean B. gingivalis and 2 kinds of type strains of B. gingivalis showed that B. gingivalis Korean-1 was reacted identically to B. gingivalis ATCC 33277. In trypsin and ${\alpha}$-glucosidase activity test of 2 Korean B. gingivalis, both of them revealed positive trypsin and negative ${\alpha}$-glucosidase activity, respectively. These investigation suggested that B. gingivalis is important pathogenic plaque bacteria for the pathogenesis of periodontitis and further study is needed to purify and characterize of the species-specific antigens of this organisms to develop monoclonal antibody and potential diagnostic reagents.

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Antimicrobial Resistance of Organisms Isolated from Clinical Specimens (임상재료에서 분리한 각종세균의 항균제내성)

  • Suh, Seong-Il;Park, Jong-Wook;Chun, Do-Ki
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.283-294
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    • 1987
  • One hundred and fifty-seven strains of staphylococci isolated from various clinical specimens and 80 of Gram-negative bacilli from urine of patients with urological diseases were tested for resistance to antimicrobial drugs by microdilution broth method. Among staphylococci, 50 to 89% of the strains were resistant to gentamicin(Gm), kanamycin(Km), erythromycin(Em), nalidixic acid(Na), and tetracycline. Ninety per cent MIC was lowest in ciprofloxacin(Cp), followed by vancomycin(Vc), trimethoprim(Tp), enoxacin(Ex), and norfloxacin(Nf) with the values of two ${\mu}g/ml$ or lower. Twenty-seven strains were resistant to methicillin(MR), with 24 strains of Staphylococcus aureus and 3 of S. epidermidis. All strains of MR S. aureus were resistant to oxacillin, rifampin(Rf), Gm, Km, Em, Na, and Tc, and no strain was resistant to Vc and Tp. Almost all staphylococci isolated from urine were S. epidermidis and sensitive to most drugs tested without MR strain. Among Gram-negative bacilli from urine, Escherichia coli(43 strains) was most frequently isolated, and followed by Klebsiella spp.(11), Proteus spp.(10), Serratia spp.(10), and Pseudomonas aeruginosa(6) in the decreasing order. The majority of E. coli and Serratia spp. were resistant to chloramphenicol(Cm), Tc, streptomycin, sulfisomidine(Su), ampicillin(Ap), Km, and carbenicillin(Cb), and 50 and 90% MICs of these drugs were also high. In Klebsiella spp., 54% or more were resistant to Cm, Su, Ap, cephalothin, and Cb. Proteus spp. were susceptible to most drugs tested, but Pseudomonas were resistant to nearly all drugs tested except Rf, amikacin, and moxalactam(Mx). All Gram-negative bacilli tested were found to be susceptible to Mx. New quinolone carboxylic acid compounds, such as Nf, Ex, and Cp showed very high antimicrobial activities against the majority of organisms tested except Pseudomonas, and 50 and 90% MICs of Nf and Ex were always equal or 2 to 4 times higher than Cp. Organisms multiply resistant to drugs were noted in almost all isolates tested. Twenty-seven strains of staphylococci were multiply resistant to 11 or more drugs, and 6 of Klebsiella spp. to 8 to 11 drugs. The most frequent multiplicity of durg resistance were 7 and 8, 12, and 13 in E. coli, Serratia spp., and Pseudomonas, respectively. No strain was resistant to more than 5 drugs in Proteus spp..

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Antimicrobial Activity of Maesil(Prunus mume) Juice against Selected Pathogenic Microorganisms (매실(Prunus mume) 착즙액의 식중독 유발균에 대한 항균 작용)

  • 이현애;남은숙;박신인
    • The Korean Journal of Food And Nutrition
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    • v.16 no.1
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    • pp.29-34
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    • 2003
  • Prunus mume has been used for the folk medicine by many old civilizations to treat food-borne diseases or enteric disorders. This study was carried out to examine the antimicrobial effect of juice from Prunus mume against pathogens such as Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Listeria monocytogenes and Bacillus cereus. The juice of Prunus mume had the strongest antimicrobial activity to Sal. enteritidis. The concentrations of Prunus mume juice for the formation of clear zone were 1% for Sai. enteritidis(15.0mm), 3% for Lis. monocytogenes(14.7mm), and 5% for Bac. cereus(14.75mm), Esc. coli(13.45mm) and Sta. aureux(11.9mm). The growth of all tested microorganisms was inhibited apparently in tryptic soy broth containing 3% and 4% Prunus mume juice. And it was found that the Prunus mume juice showed the highest antimicrobial properties, followed by Sal. enteritidis, Bac. cereus, Sta. aureus, Lis. monocytogenes, Esc. coli.