• Title/Summary/Keyword: Inoculum concentration

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Optimization of Cu, Hg and Cd removal by Enterobacter cloacae by ferric ammonium citrate precipitation

  • Singh, Rashmi R.;Tipre, Devayani R.;Dave, Shailesh R.
    • Advances in environmental research
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    • v.3 no.4
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    • pp.283-292
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    • 2014
  • Iron precipitating organisms play a significant role in the formation of ferric hydroxide precipitate, which acts as strong adsorbent for toxic metal. In this respect four different iron precipitating cultures were isolated from Hutti gold mine surface winze water sample on citrate agar medium. The best isolate was screened out for metal removal study on the basis of fast visual iron precipitation. The selected isolate was identified as Enterobacter sp. based on routine biochemical tests and Biolog GN microplate results and as Enterobacter cloacae subsp. dissolvens by 16S rRNA gene sequence analysis (GenBank accession number EU429448). Influence of medium composition, medium initial pH, the influence of inoculum size, effect of various media and ferric ammonium citrate concentration were studied on metal removal in shake flask experiments. Under the optimized conditions studied, E. cloacae showed $94{\pm}2$, $95{\pm}2$ and $70{\pm}2%$ of cadmium, copper and mercury removal from a simulated waste in shake flask studies. In lab scale column reactor more than 85% of copper and mercury removal was achieved.

Studies On Induction of ${\beta}$-D-galactosidase In Candida kefyr (Candida kefyr의 ${\beta}$-D-galactosidase 合成誘導에 關한 硏究[I])

  • Chun, Soon-Bai
    • Korean Journal of Microbiology
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    • v.22 no.2
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    • pp.77-84
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    • 1984
  • This examined some conditions for the induction of ${\beta}$-D-galactosidase synthesis in Candida kefyr CBS 834. The optimal pH, temperature, and inoculum size either for growth or${\beta}$-D-galactosidase synthesis were 5.5, $30^{\circ}C$ and above 0.2 at A610nm, respectively. Enzyme activity began to increase at 2h after the addition of inducer, and continued to increase linearly up to $2{\sim}3h$ before reaching stationary phase, and thereafter its activity was decreased. ${\beta}$-D-galactosidase was induced either by lactose or galactose but not either by glucose or ethanol. The greater activity of ${\beta}$-D-galactosidase on galactose than on lactose indicated that the former might be natural inducer for ${\beta}$-D-galactosidase synthesis. The rate of its induction as a function of lactose concentration showed that enzyme activity increased linearly above 4mM, while it was very low below that. Glucose represed the induction of ${\beta}$-D-galactosidase, and the period of adaptation to inducer from other carbon sources was relatively short.

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Fungichromin Production by Streptomyces padanus PMS-702 for Controlling Cucumber Downy Mildew

  • Fan, Ya-Ting;Chung, Kuang-Ren;Huang, Jenn-Wen
    • The Plant Pathology Journal
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    • v.35 no.4
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    • pp.341-350
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    • 2019
  • Streptomyces padanus PMS-702 strain produces a polyene macrolide antibiotic fungichromin and displays antagonistic activities against many phytopathogenic fungi. In the present study, experimental formulations were assessed to improve the production of fungichromin, the efficacy of PMS-702 on the suppression of sporangial germination, and the reduction of cucumber downy mildew caused by Pseudoperonospora cubensis. PMS-702 strain cultured in a soybean meal-glucose (SMG) medium led to low levels of fungichromin accumulation and sporangial germination suppression. Increasing medium compositions and adding plant oils (noticeably coconut oil) in SMG significantly increased fungichromin production from 68 to $1,999.6{\mu}g/ml$. Microscopic examination reveals that the resultant suspensions significantly reduced sporangial germination and caused cytoplasmic aggregation. Greenhouse trials reveal that the application of PMS-702 cultural suspensions reduced downy mildew severity considerably. The addition of Tween 80 into the synthetic medium while culturing PMS-702 further increased the suppressive efficacy of downy mildew severity, particularly when applied at 24 h before inoculation or co-applied with inoculum. Fungichromin at $50{\mu}g/ml$ induced phytotoxicity showing minor necrosis surrounded with light yellowish halos on cucumber leaves. The concentration that leads to 90% inhibition (IC90) of sporangial germination was estimated to be around $10{\mu}g/ml$. The results provide a strong possibility of using the S. padanus PMS-702 strain as a biocontrol agent to control other plant pathogens.

Kinetic Study of the Anaerobic Digestion of Swine Manure at Mesophilic Temperature: A Lab Scale Batch Operation

  • Kafle, Gopi Krishna;Kim, Sang-Hun
    • Journal of Biosystems Engineering
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    • v.37 no.4
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    • pp.233-244
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    • 2012
  • Purpose: The kinetic evaluation was performed for swine manure (SM) degradation and biogas generation. Methods: The SM was anaerobically digested using batch digesters at feed to inoculum ratio (F/I) of 1.0 under mesophilic conditions ($36.5^{\circ}C$). The specific gas yield was expressed in terms of gram total chemical oxygen demand (mL/g TCOD added) and gram volatile solids added (mL/g VS added) and their effectiveness was discussed. The biogas and methane production were predicted using first order kinetic model and the modified Gompertz model. The critical hydraulic retention time for biomass washout was determined using Chen and Hashimoto model. Results: The biogas and methane yield from SM was 346 and 274 mL/ TCOD added, respectively after 100 days of digestion. The average methane content in the biogas produced from SM was 79% and $H_2S$ concentration was in the range of 3000-4108 ppm. It took around 32-47 days for 80-90% of biogas recovery and the TCOD removal from SM was calculated to be 85%. When the specific biogas and methane yield from SM (with very high TVFA concentration) was expressed in terms of oven dried volatile solids (VS) basis, the gas yield was found to be over estimated. The difference in the measured and predicted gas yield was in the range of 1.2-1.5% when using first order kinetic model and 0.1% when using modified Gompertz model. The effective time for biogas production ($T_{Ef}$) from SM was calculated to be in the range of 30-45 days and the critical hydraulic retention time ($HRT_{Critical}$) for biomass wash out was found to be 9.5 days. Conclusions: The modified Gompertz model could be better in predicting biogas and methane production from SM. The HRT greater than 10 days is recommended for continuous digesters using SM as feedstock.

Factors Affecting on the Production of Exopolysaccharides from Zoogloea ramigera l15SLR. (Zoogloea ramigera l15SLR의 다당류 생산에 영향을 미치는 인자)

  • 소한섭;김찬식;이삼빈
    • Microbiology and Biotechnology Letters
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    • v.30 no.3
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    • pp.264-269
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    • 2002
  • The production of exopolysaccharide from Zoogloea ramigera 115SLR in batch culture was affected by carbon sources, rifampicin concentration, inoculum size and cell density of starter culture. The increase of organic nitrogen concentration and cell density in defined medium by adding starter culture resulted in higher production of exopolysaccharide (EPS) within 2 days. Glucose and lactose as carbon sources showed EPS production of 10.7 gk and 10.5 ga, respectively. Higher EPS production was obtained with 2.5% (w/v) glucose. White sugar, brown sugar and galactose produced less than 10 gt of EPS. The optical density of starter culture affected on EPS yield, producing over 10 gt of EPS in the range of 1.0-1.7. In the presence of rifsmpicin, Z. ramigera 115SLR produced EPS of 12.9 gt. Molecular weight of EPS produced with/without rifampicin was determined with 1.367$\times$106 and 1.711$\times$106 g/mol using HPSEC-MALLS-RI, respectively.

Analysis of Optimum Condition for Alcoholic Drink Production Using Onion Extract. (양파즙을 사용한 알코올 음료 제조를 위한 최적조건 검토)

  • Kim, Sam-Woong;Oh, Eun-Hye;Jun, Hong-Ki
    • Journal of Life Science
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    • v.18 no.6
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    • pp.871-877
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    • 2008
  • Onions are considered to be a promising source of the alcoholic drink because they are rich in sugar, amino acids and various nutrients. To isolate strains of Saccharomyces cerevisiae producing ethanol of higher concentration, 19 strains were subjected to screening. Among them, the strain producing the highest concentration of ethanol was OJ-8 strain. Onion's oder was effectively removed by treatment for 30 min with 10% (w/v) charcoal against medium and then heat treatment of onion extract for 40 min at $100^{\circ}C$. The optimum conditions for alcoholic fermentation was investigated in medium containing the onion extract. The optimal conditions for ethanol production was obtained by standing culture for 5 days at $25^{\circ}C$ with 5% inoculum volume.

Evaluation and Optimization of a Serum-based Minimum Inhibitory Concentration Assay to Caspofungin in Candida albicans Clinical Isolates

  • Yoo, Young Bin;Kim, Sung-Soon;Kim, Young Kwon;Kim, Sunghyun
    • Biomedical Science Letters
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    • v.22 no.4
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    • pp.174-183
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    • 2016
  • In the present study, a serum-based minimum inhibitory concentration (MIC) testing to caspofungin was optimized and evaluated to solve the limitations of the conventional Clinical and Laboratory Standards Institute (CLSI) guideline-based antifungal agent MIC test and the usefulness of this testing for clinical application was determined. A total of 105 Candida albicans clinical isolates were used for measuring MIC to caspofungin. Results showed that growth characteristics were different according to types of serum and the mouse serum was the most suitable for this assay. In order to measure the optimal concentration of mouse serum, 0 to 100% mouse serum were added to the media during fungal culture. The optimal concentration of serum was 50% when consideration of antifungal agent administration and inoculum size, serum components and ease of hyphae separated, and the consideration of the degree of growth. In comparison of the usefulness between the conventional Alamar-modified broth microdilution MIC assay and 50% mouse serum-based MIC testing, the range of $MIC_{80}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.42{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $2.0{\sim}32.0{\mu}g/mL$ (SD ${\pm}9.01{\mu}g/mL$). The range of $MIC_{50}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.40{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $1.0{\sim}16.0{\mu}g/mL$ (SD ${\pm}2.36{\mu}g/mL$). The MICs of 50% mouse serum-based MIC testing was increased by up to 4 to 64 times than Alamar-modified broth microdilution MIC assay. In conclusion, a 50% mouse serum-based MIC assay was more useful for measuring MIC in Candida albicans clinical isolates than conventional colorimetric broth microdilution MIC testing.

Involvement of Growth-Promoting Rhizobacterium Paenibacillus polymyxa in Root Rot of Stored Korean Ginseng

  • Jeon, Yong-Ho;Chang, Sung-Pae;Hwang, In-Gyu;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.881-891
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    • 2003
  • Paenibacillus polymyxa is a plant growth-promoting rhizobacterium (PGPR) which can be used for biological control of plant diseases. Several bacterial strains were isolated from rotten roots of Korean ginseng (Panax ginseng C. A. Meyer) that were in storage. These strains were identified as P. polymyxa, based on a RAPD analysis using a P. polymyxa-specific primer, cultural and physiological characteristics, an analysis utilizing the Biolog system, gas chromatography of fatty acid methyl esters (GC-FAME), and the 16S rDNA sequence analysis. These strains were found to cause the rot in stored ginseng roots. Twenty-six P. polymyxa strains, including twenty GBR strains, were phylogenetically classified into two groups according to the ERIC and BOX-PCR analyses and 16S rDNA sequencing, and the resulting groupings systematized to the degrees of virulence of each strain in causing root rot. In particular, highly virulent GBR strains clustered together, and this group may be considered as subspecies or biovar. The virulence of the strains seemed to be related to their starch hydrolysis enzyme activity, but not their cellulase or hemicellulase activity, since strains with reduced or no starch-hydrolytic activity showed little or no virulence. Artificial inoculation of the highly virulent strain GBR-1 onto the root surfaces of Korean ginseng resulted in small brown lesions which were sunken and confined to the outer portion of the root. Ginseng root discs inoculated in vitro or two-year-old roots grown in soil drenched with the inoculum developed significant rot only when the inoculum density was $10^{6}-10^{7}$ or more colony-forming units (CFU) per ml. These results suggest that P. polymyxa might induce ginseng root rot if their population levels are high. Based on these results, it is recommended that the concentration of P. polymyxa should be monitored, when it is used as a biocontrol agent of ginseng, especially in the treatment of stored roots.

Improvement of Peroxidase Productivity by Optimization of Medium Composition and Cell Inoculum Size in Suspension Cultures of Sweet Potato (Ipomoea batatas) (고구마(Ipomoea batatase)현탁배양에서 배지조성 및 세포접종량의 적정화에 의한 Pemxidase생산성 향상)

  • 곽상수;김수경;정경희;유순희;박일현;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.91-97
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    • 1994
  • To improve the productivity of peroxidase (POD) of cell line SP-47 derived from cell suspension cultures of sweet potato (Ipomoea batatas (L) Lam.cv White Star), we optimized culture conditions including the composition and concentration of plant growth regulators and carbon source, and the cell inoculum size. When one g (fr wt) of cells was inoculated into 50 mL TL medium supplemented with l mg/L 2,4-D and 30g/L sucrose in 300 mL Erlenmeyer flask at 25$^{\circ}C$ in the dark (100rpm), the POD activity per g cell dry wt was maximized to be about 6,800 units after 25 days of subculture, which was about 30 times higher than that of intact roots of horseradish plants grown in the greenhouse, but the cell growth was maximum after 15 days of subculture. The protein content per g cell dry wt maintained almost plateau and after 25 days of subculture decreased as culture Proceeded further whereas the POD specific activity (unit/mg protein) was about two times higher after subculture and continuously increased from 12 days to the end of cultures (40 days). The POD isozyme patterns showed almost the same regardless of cell growth stage, but some acidic isozymes were slightly increased after 25 days of subculture. These results indicate that POD activity in suspension cultures of sweet potato is closely associated with cell growth and stresses derived from cell culture renditions and medium depletion. Due to its high POD activity the SPL47cell line seems to be suitable for the mass production of POD.

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Development of Effective Screening Method and Evaluation of Radish Cultivars for Resistance to Fusarium oxysporum f. sp. raphani (효율적인 무 시들음병 저항성 검정법 개발 및 무 품종들의 병 저항성 평가)

  • Baik, Song-Yi;Kim, Jin-Cheol;Jang, Kyoung-Soo;Choi, Yong-Ho;Choi, Gyung-Ja
    • Research in Plant Disease
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    • v.16 no.2
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    • pp.148-152
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    • 2010
  • To establish the efficient screening method for resistance of radish to F. oxysporum f. sp. raphani, we investigated the development of Fusarium wilt of two radish cultivars, 'Songbaek' (susceptible) and 'Tokwang' (moderately resistant), according to several conditions such as inoculation methods, inoculum concentrations, and dipping periods of radish roots in spore suspension. By infected soil and soil-drenching inoculation methods, Fusarium wilt did not occur on the seedlings of both cultivars. In root dipping inoculation method using cut or non-cut roots of radish plants, the cut roots were easily infected by the pathogen than non-cut roots. And the disease development of two cultivars represented significant difference in non-cut root method. On the other hand, disease severity of Fusarium wilt on radish seedlings according to inoculum concentration increased in a dose-dependant manner, regardless of dipping periods. Using screening method established from the results, the 41 commercial radish cultivars were evaluated the degree of resistance to F. oxysporum f. sp. raphani. Among them, 6 radish cultivars were resistant, 22 cultivars were moderately resistant, and 13 cultivars were susceptible to Fusarium wilt.