• Title/Summary/Keyword: Co-incubation

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Degradation Characteristics of Paper Sludge and Changes of Heavy Metals in Soil (토양중 제지슬러지의 분해 특성 및 중금속 변화)

  • Lee, Hong-Jae;Jeong, In-Ho;Cho, Ju-Sik;Heo, Jong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.15 no.3
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    • pp.296-305
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    • 1996
  • Chemical characteristics of paper sludge, degradation of the sludge in soil and $CO_2$ generation, and changes of nitrogen and heavy metals in soil treated with the sludge were investigated. The results obtained was summarized as follows: 1. Degradation rate of paper sludge in soil was 19% at room temperature, and 28% at $incubation(30^{\circ}C)$ temperature after 12-weeks treatment. 2. T-C, T-N and the C/N ratio of the sludge in soil at room temperature were 15.5%, 0.22% and 71 respectively, and 14.5%, 0.24% and 60, respectively, at $incubation(30^{\circ}C)$ temperature after 12-week treatment. 3. $CO_2$ genaration in soil treated with 1%, 3% and 5% of the sludge was 247mg/100g, 334mg/100g and 458mg/100g, respectively, at room temperature, and 385mg/100g, 550mg/100g and 618mg/100g, respectively, at incubation temperature after 12 weeks treatment. 4. Mineralization ratio of organic nitrogen in soil treated with 1%, 3% and 5% of the sludge was 8.7%, 13.4% and 16.2%, respectively, at $incubation(30^{\circ}C)$ temperature after 12-weeks treatment. 5. The amounts of DTPA-extractable Cu, Cd, Zn, Pb, and Cr in Soil treated with paper sludge were $0.7{\sim}2.2$, $0.1{\sim}0.17$, $1.4{\sim}2.8$, $1.4{\sim}2.8$, and $0{\sim}0.7mg/kg$, respectively. Mean while, those of $HNO_3$ extractable Cu, Cd, Zn, Pb, and Cr were $7.9{\sim}10.0$, $0.6{\sim}0.9$, $17.6{\sim}34.4$, $14.7{\sim}18.5$, and $5.8{\sim}9.0mg/kg$, respectively.

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Effects of the Processing Methods of Forage Rye (Secale cereale) on Rumen Fermentation Characteristics and Greenhouse Gas Emissions In Vitro of Hanwoo (호밀 조사료 가공방법이 한우의 반추위 내 발효특성과 온실가스 발생량에 미치는 영향)

  • Ji Yoon Kim;Seung Min Jeong;Young Ho Joo;Chang Hyun Baeg;Bu Gil Choi;Arrynda Rachma Dyasti Wardani;Sam Churl Kim
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.44 no.2
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    • pp.99-105
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    • 2024
  • This study was conducted to estimate the effects of the forage process on rumen fermentation characteristics and greenhouse gas emissions of rye. Rye was grown at the Taeyoung Livestock farm and harvested at the heading stage. The harvested rye (5 kg) was sub-sampled for fresh forage, hay, and silage in triplicates. The sub-sampled rye was freeze-dried or air-dried for fresh forage or rye hay, respectively. For rye silage, the sub-sampled rye forage was ensiled into a 10 L mini bucket silo and stored for 90 days. For 72 h rumen incubation, each forage (0.3 g) was placed into the incubation bottle with the rumen mixture (30 mL) in quadruplicates. After the incubation, total gas was measured and sub-sampled for CO2 and CH4 analyses, and the bottle content was centrifuged for in vitro digestibilities of dry matter (IVDMD) and neutral detergent fiber (IVNDFD), and rumen fermentation characteristics. Silage had higher crude protein, crude ash, and acid detergent fiber concentrations than fresh forage and hay but lower non-fiber carbohydrates and relative feed value (p<0.05). And, silage had higher lactic acid bacteria than the other forages but lower pH (p<0.05). After 72 h incubation in the rumen, fresh forage had higher IVDMD and butyrate content than the other forages (p<0.05). However, silage had higher rumen pH and propionate content than the other forages but lower A:P ratio (p<0.05). Regarding greenhouse gases, silage had lowest total gas (mL/g DMD and NDFD) and CH4 (mL/g DMD and NDFD) emissions, while fresh forage had lowest CO2 (mL/g DMD) emission (p<0.05). Therefore, this study concluded that the ensiling process of rye can effectively mitigate greenhouse gas emissions of Hanwoo.

In vitro Antibacterial Activities of Novel Fluoroquinolone DWP20367 (신합성 플로로퀴놀론계 항생물질인 DWP20367의 In vitro 항균효과)

  • Kim, Ji-Yeon;Choi, Moon-Jung;Han, Seung-Hee;Shim, Jeom-Soon;Jung, Yeon-Eui;Son, Ho-Jung;Lee, Jae-Wook;Yu, Young-Hyo;Park, Myung-Hwan
    • YAKHAK HOEJI
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    • v.41 no.2
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    • pp.225-232
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    • 1997
  • The in vitro antibacterial activity of DWP20367 (1-Cyclopropyl-6-fluoro-8-chloro-7-(2,7-diazabicyclo[3,3,0]oct-4-ene-7-yl)-1,4-dihydro-4-oxoquinoline-3-carboxylic acid), a new broad-spectrum fluoroquinolone, was compared with those of ciprofloxacin (CPFX), sparfloxacin (SPFX) and ofloxacin (OFLX). DWP20367 was showed antibacterial activity much higher than CPFX, SPFX and OFLK against gram-positive bacteria, while it was slightly more superior to quinolones against gram-negative bacteria. DWP20367 was particularly effective against MRSA, and its $MlC_{90}$ against clinical isolates of methicillin-susceptible S. aureus, low methicillin-resistant S. aureus and high methicillin-resistant S. aureus were 0.098, 0.781 and 1.563 micro g/ml, respectively. Against S. pneumoniae, MIC90 of DWP20367 was 2- to 8-fold higher than those of CPFX. With a view of MIC90, DWP20367 showed slightly more potent activity against P. aeruginosa and E. coli isolates than the control quinolones. DWP20367 activity was not affected by inoculum size and medium pH. But addition of $Mg^{2+}, \;Ca^{2+} $Mg2+, Ca2+ or horse serum (25%) decreased its antibacterial activity. DWP20367 was showed rapidly bactericidal activity within 2 to 4 h, and regrowth was not observed even after 24 h incubation at concentrations near the 4-fold of MIC.

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Development of a Functional Mixed-Starter Culture for Kefir Fermentation (Kefir 배양용 기능성 복합 Starter 개발)

  • Lee, Bomee;Yi, Hae-Chang;Moon, Yong-II;Oh, Sejong
    • Journal of Dairy Science and Biotechnology
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    • v.36 no.3
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    • pp.178-185
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    • 2018
  • Kefir, which originates in the Caucasian mountains, is a cultured milk beverage produced by a combination of acidic and alcoholic fermentation. Kefir products are commonly used as food vehicles to deliver health-promoting materials including kefran and lactic acid bacteria to consumers. The aim of this study was to develop a freeze-dried starter culture without yeast and assess the suitability of kefir-like dairy products for the growth of lactic acid bacteria and the acidification of milk. Pasteurized whole milk (SNF 8.5%) stored at $25^{\circ}C$ was aseptically inoculated with starter cultures (0.002% w/v); it was kept at $25^{\circ}C$ until the pH attained a value of 4.6. Ten grams of the kefir-like product sample was diluted with 90 mL of 0.15% peptone water diluent in a milk dilution bottle, followed by uniform mixing for 1 min. Viable cells of Lactobacillus species were enumerated on modified-MRS agar (pH 5.2), with incubation at $37^{\circ}C$ for 48 h. Viable cells of Lactococcus species were enumerated on M17-lactose agar, with incubation at $32^{\circ}C$ for 48 h. The pH attained a value of 4.6 after fermentation for 9 h 30 min (Starter 1), 9 h 45 min (Starter 2), and 12 h (Starter 3). The viable cell count of Lactobacillus sp. and Lactococcus sp. was initially $10^5{\sim}10^6CFU/g$; it increased significantly to $10^9CFU/g$ after 12 h of incubation. During the storage of the kefir-like products at $4^{\circ}C$ for 1 4 days, the total viable cell numbers were unchanged, but the pH decreased slightly. The consistency of the kefir products increased gradually during the storage. The organoleptic properties of the kefir products fermented using the new starter culture are more desirable than those of commercial kefir. These results suggest that the newly developed starter culture without yeast could be suitable for kefir fermentation.

Influence of Increased Carbon Dioxide Concentration on the Bioluminescence and Cell Density of Marine Bacteria Vibrio fischeri (이산화탄소 농도 증가에 따른 발광미생물의 상대발광량과 밀도변화에 대한 연구)

  • Sung, Chan-Gyoung;Moom, Seong-Dae;Kim, Hye-Jin;Choi, Tae-Seob;Lee, Kyu-Tae;Lee, Jung-Suk;Kang, Seong-Gil
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.15 no.1
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    • pp.8-15
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    • 2010
  • An experiment was conducted to evaluate the biologically adverse effect of increased carbon dioxide in seawater on marine bacteria, Vibrio fischeri. We measured the bioluminescence and cell density at every 6 hours for 24 hours of the whole incubation period after exposing test microbes to a range of $CO_2$ concentration such as 380(Control), 1,000, 3,000, 10,000 and 30,000 ppm, respectively. Significant effect on relative luminescence(RLU) of V. fischeri was observed in treatments with $CO_2$ concentration higher than 3,000 ppm at t=12 h. However, the difference of RLU among treatments significantly decreased with the incubation time until t=24 h. Similar trend was observed for the variation of cell density, which was measured as optical density using spectrophotometer. The results showed that a significant relationship between $CO_2$ concentration and bioluminescence of test microbes was observed for the mean time. However, the inhibition of relative bioluminescence and also cell density could be recovered at the concentration levels higher than 3,000 ppm. The dissolved $CO_2$ can be absorbed directly by cell and it can decrease the intracellular pH. Our results implied that microbes might be adversely affected at the initial growing phase by increased $CO_2$. However, they could adapt by increasing ion transport including bicarbonate and then could make their pH back to normal level. Results of this study could be supported to understand the possible influence on marine bacteria by atmospheric increase of $CO_2$ in near future and also by released $CO_2$ during the marine $CO_2$ sequestration activity.

Improvement of Pregnancy Rates by Coculture of Human Embryos with Cumulus Cells in Glucose and Phosphate Free M-TALP Media (Glucose와 Phosphate가 제거된 M-TALP 배지에서의 난구세포 공배양에 의한 임신율 향상에 관한 연구)

  • Chung, B.S.;Chang, W.H.;Lee, M.H.;Kim, J.Y.;Bang, J.H.;Kim, K.H.;Suh, T.K.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.75-81
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    • 1999
  • The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.

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In vivo and In vitro Metabolism of Recombinant Human Epidermal Growth Factor (DWP401) in Rats (재조합 인간 상피세포성장인자(DWP401)의 흰쥐에서의 in vivo와 in vitro 대사)

  • Koh, Yeo-Wook;Nam, Kouen-Ho;Jung, Ju-Young;Park, Seung-Kook;Yu, Young-Hyo;Kim, Jae-Hwan;Han, Kun;Park, Myung-Hwan;Shim, Chang-Koo
    • YAKHAK HOEJI
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    • v.41 no.3
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    • pp.381-388
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    • 1997
  • Metabolism of DWP401, recombinant juman epidermal growth factor, was examined in vivo and in vitro in rats. When $^{125}I$-labeled DWP401 was administered at a dose of 50 ${\mu}g$/kg by i.v. injection. $^{125}I$-labeled DWP401 was rapidly degraded within 30 minytes above 93%. Thin layer chromatography analysis of urine collected for 24 hr after i.v. administration of $^{125}I$-labeled DWP401 showed ohly one spot on a X-ray film which was considered as diiodo-tyrosine. This result suggests tha $^{125}I$-labeled DWP401 was completely digested into free amino acids without any specific intermediate polypeptides. About 42.1% of the administered iodine was recovered in 24 hr. For in vitro degradation study, $^{125}I$-labeled DWP401 was added to plama and tissue homogenates of rats and incubated at $37^{\circ}C$. Almost 98% of the added radioactivity recovered from the protein fraction of the liver, kidey, small intestine, stomach and spleen decreased rapidly. For examplem the recovery rates of $^{125}I$-labeled DWP401 were 58.6, 63.2, 39.9, 52.9 and 66.8% after 4hrs of incubation in respective organ homogenates.

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In vitro Antibacterial Activity of DWP20418, a New Carbapenem Antibiotic (새로운 카바페넴계 항생물질인 DWP20418의 In vitro 항균작용)

  • Kim, Ji-Yeon;Choi, Moon-Jung;Park, Nam-Joon;Yim, Seong-Soo;Byun, Young-Seok;Yu, Young-Hyo;Park, Myung-Hwan
    • YAKHAK HOEJI
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    • v.41 no.2
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    • pp.233-240
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    • 1997
  • The in vitro antibacterial activity of DWP20418 (1R, 5S, 6S)-6-[1-(R)-Hydroxyethyl)-l-methyl-2-[(2S,4S)-2-(piperazinylcarbonyl)-1-(R)-hydroxyethyl)pyrrolidine-4-thio]carb apen-2-em-3-carboxylic acid), a new carbapenem antibiotic, was compared with those of imipenem (IPM) and meropenem (MEPM). DWP20418 was comparable or slightly more superior to MEPM against gram-positive bacteria, and it showed more potent activity to IPM against gram-negative bacteria. DWP20418 was particularly active against MRSA, and its $MIC_{90}$ of methicillin-susceptible S. aureus, low methicillin-resistant S. aureus and high methicillin-resistant S. aureus were 0.391, 25 and 50 ${\mu}g/ml$, respectively. With a view of $MIC_{90}$, DWP20418 was comparable than the other carbapenems against P. aeruginosa and E. coli isolates. The activity of DWP20418 was not affected in the presence of $Mg^{2+},\;Ca^{2+}$ or horse serum. But inoculum size and alterations in pH of medium affected its antibacterial activity. DWP20418 showed rapidly bactericidal activity within 1h, and regrowth was not observed even incubation of 24hrs at the concentrations near the MIC.

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Kinetic Responses of Soil Carbon Dioxide Emission to Increasing Urea Application Rate

  • Lee, Sun-Il;Lim, Sang-Sun;Lee, Kwang-Seung;Kwak, Jin-Hyeob;Jung, Jae-Woon;Ro, Hee-Myoung;Choi, Woo-Jung
    • Korean Journal of Environmental Agriculture
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    • v.30 no.2
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    • pp.99-104
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    • 2011
  • BACKGROUND: Application of urea may increase $CO_2$ emission from soils due both to $CO_2$ generation from urea hydrolysis and fertilizer-induced decomposition of soil organic carbon (SOC). The objective of this study was to investigate the effects of increasing urea application on $CO_2$ emission from soil and mineralization kinetics of indigenous SOC. METHODS AND RESULTS: Emission of $CO_2$ from a soil amended with four different rates (0, 175, 350, and 700 mg N/kg soil) of urea was investigated in a laboratory incubation experiment for 110 days. Cumulative $CO_2$ emission ($C_{cum}$) was linearly increased with urea application rate due primarily to the contribution of urea-C through hydrolysis to total $CO_2$ emission. First-order kinetics parameters ($C_0$, mineralizable SOC pool size; k, mineralization rate) became greater with increasing urea application rate; $C_0$ increased from 665.1 to 780.3 mg C/kg and k from 0.024 to 0.069 $day^{-1}$, determinately showing fertilizer-induced SOC mineralization. The relationship of $C_0$ (non-linear) and k (linear) with urea-N application rate revealed different responses of $C_0$ and k to increasing rate of fertilizer N. CONCLUSION(s): The relationship of mineralizable SOC pool size and mineralization rate with urea-N application rate suggested that increasing N fertilization may accelerate decomposition of readily decomposable SOC; however, it may not always stimulate decomposition of non-readily decomposable SOC that is protected from microbial decomposition.

Effect of Co-Culture Mouse Fetal Fibroblast Cell on In Vitro Development of Blastomeres Separated from Mouse Preimplantation Embryos (생쥐 태아 Fibroblast 세포와 공동배양이 초기 생쥐배 분할구의 체외 발생능에 미치는 영향)

  • 김진호;정병헌;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.16 no.4
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    • pp.341-346
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    • 1993
  • The development of isolated blastomeres from mammalian preimplantation embryos has been basically studied for the multiplication of embryos from superior animals. Therefore, this study was investigated the effect of co-culture with mouse fetal fibroblast cells(MFFC) on in vitro development of blastomeres from mouse preimplantation embryos. Mature female ICR mice were treated with hormone to induce superovulation and embryos were collected at each 2, 4, and 8-cell stage. Then, after removing zona pellucida with protease, blastomeres were isolated by micropipetting, or reconstituted with different stage blastomere, and incubated for 72 hrs either in T6 or TCM199 or on the monolayer of MFFC, which was prepared with fibroblast cells from 14∼14 day mouse fetus. After incubation, we examined their development rates every day and the nuclei numbers of each blastocyst by Hoechst-33342 staining. In the development rates of blastomeres, there were no significant differences between media but the higher rateswere found in the monolayer of MFFC, regardless of reconsititution. In addition, blastomeres cultured with MFFC had slightly greater number of nuclei than those cultured in single media. Generally, the higher development rates of blastomeres were found from earlier stage embryos than the later ones, regardless of culture conditions. Reconsitituted blastomeres had more nuclei but did not show the higher development rates, compared to the single blastomeres. Taken together, our results suggest that co-culture with MFFC have a beneficial effect on the in vitro development of blastomeres from mouse embryos.

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