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Comparison of Culture Media for In Vitro Maturation of Oocytes of Indigenous Zebu Cows in Bangladesh

  • Singha, Joydev Kumar;Bhuiyan, Mohammad Musharraf Uddin;Rahman, Mohammad Moshiur;Bari, Farida Yeasmin
    • 한국수정란이식학회지
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    • 제30권4호
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    • pp.327-333
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    • 2015
  • The objectives of the present study were to select an effective basic medium including its hormone and protein supplementation for IVM of oocytes of indigenous zebu cows. The ovaries of cows were collected from slaughter house and the follicular fluid was aspirated from 2 to 8 mm diameter follicles. The COCs with more than 3 cumulus cell layers and homogenous cytoplasm were selected for maturation. The oocytes were matured in media for 24 hrs at $39^{\circ}C$ with 5% $CO_2$ in humidified air. The maturation of oocytes was evaluated by examining the presence of first polar body under microscope. An efficient basic medium was determined after culturing COCs in either TCM 199 or SOF medium in Experiment 1. An efficient hormone supplementation was determined after culturing COCs in either FSH or gonadotrophin supplemented TCM 199 in Experiment 2. An efficient protein supplementation was determined after culturing COCs in either FBS or Oestrous cow serum (OCS) supplemented TCM 199 in Experiment 3. The oocyte recovery rate per ovary was 3.35. The overall rate of IVM was 74.6%. The maturation rate was $75.5{\pm}3.9$ and $62.2{\pm}20.2%$ in TCM and SOF medium, respectively (P>0.05). The maturation rate of oocytes was significantly higher ($76.6{\pm}13.2%$) in FSH supplemented medium than gonadotrphin supplemented counterpart ($69.7{\pm}10.8%$) (P<0.05). The maturation rates of oocytes were $81.7{\pm}12.9$ and $85.7{\pm}12.7%$ in medium supplemented with FBS and OCS, respectively (P>0.05). In conclusions, both TCM 199 and SOF supplemented with either FBS or OCS, and FSH may be used as medium for IVM of indigenous zebu oocytes in Bangladesh.

Gluten-Free 쌀빵의 제빵 특성에 대한 첨가 수분 온도 영향 (Effect of Added Water Temperature on Baking Characteristics of Gluten-Free Rice Bread)

  • 김상숙;정혜영
    • 한국식품영양학회지
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    • 제30권6호
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    • pp.1184-1190
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    • 2017
  • Baking characteristics of gluten-free rice bread were investigated, when 20, 30, 40, 50 and $60^{\circ}C$ water was added during mixing. The temperature of the dough before fermentation was affected by the temperature of the water and the mixing time. When $60^{\circ}C$ water was added, the specific gravity of the dough was the highest before fermentation (p<0.01). The specific gravity of the dough after fermentation was 32~39% of the specific gravity of the dough before fermentation. When $50^{\circ}C$ water was added, the volume and the specific volume of rice bread were higher than those in addition of water at other temperatures (p<0.001). In case of adding water of $50^{\circ}C$, the shape of the rice bread showed the largest volume, high appearance and a round shape. After storage for 2 and 24 hours, the addition of water of $50^{\circ}C$ resulted in the lowest hardness and chewiness values of rice bread. The sensory descriptive analysis revealed that when $50^{\circ}C$ water was added, the air cell size, springiness and hardness values of gluten-free rice bread were lower than those in addition of water at other temperatures. There was a difference in the appearance and texture of gluten-free rice bread, when 20, 30, 40, 50 and $60^{\circ}C$ water was added during mixing.

암모니아 센서를 이용한 간헐폭기 Membrane bioreactor공정에서의 전력비 저감과 관형막을 이용한 슬러지 농축에 관한 연구 (A study on an intermittent aeration membrane bioreactor system using ammonia sensor to decrease energy consumption and sludge concentration by tubular membrane)

  • 강희석;이의종;김형수;장암
    • 상하수도학회지
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    • 제28권2호
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    • pp.161-170
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    • 2014
  • It is essential to decrease energy consumption and excess sludge to economically operate sewage treatment plant. This becomes more important along with a ban on sea dumping and exhaustion of resource. Therefore, many researchers have been study on energy consumption reduction and strategies for minimization of excess sludge production from the activated sludge process. The aeration cost account for a high proportion of maintenance cost because sufficient air is necessary to keep nitrifying bacteria activity of which the oxygen affinity is inferior to that of heterotrophic bacteria. Also, additional costs are incurred to stabilize excess sludge and decrease the volume of sludge. There were anoxic, aerobic, membrane, deairation and concentration zone in this MBR process. Continuous aeration was provided to prevent membrane fouling in membrane zone and intermittent aeration was provided in aerobic zone through ammonia sensor. So, there was the minimum oxygen to remove $NH_4-N$ below limited quantity that could be eliminated in membrane zone. As the result of this control, energy consumption of aeration system declined by between 10.4 % and 19.1 %. Besides, we could maintain high MLSS concentration in concentration zone and this induced the microorganisms to be in starved condition. Consequentially, the amount of excess sludge decrease by about 15 %.

체외배양기술로 생산된 초기배에 의한 한우 송아지 생산기술 III. 배반포와 발생에 미치는 발생배지의 효과와 동결융해후의 생존율에 미치는 Sucrose와 Trehalose의 효과 (Development of Production Techniques for Korean Native Cattle Calves from Embryos by In Vitro Technology 3. Effects of Culture Medium on Blastocyst Development and Effects of Sucrose and Trehalose on the Survival Rate of In Vitro Developed Embryos after Thawing)

  • 서경덕;김호중;김갑수;김광식
    • 한국수정란이식학회지
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    • 제13권2호
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    • pp.173-178
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    • 1998
  • The objective of this study was to evaluate the embryonic development ability and the appearance of blastocysts of bovine in vitro fertilized oocytes cultured in different culture media, and also to evaluate survival rate after thawing of frozen embryos by using 1.5 or 1.8M ethylene glycol(EG) with sucrose or trehalose. Fertilized oocytes were divided into three groups; i ) monolayer of cumulus /granulosa cell prepared by TGM 199+5% calf serum(TGM199), ii)GRlaa+5% CS, iii)SOF+5% CS, and they were cultured after insemination for 9 days, at 39˚C, under 5% $CO_2$ in air, but SOF+5% CS was cultured at 39˚C, under 5% 02, 5% GO2, 99% N2. Blastocysts derived from GRlaa + 5% CS on day 7~8 after insemination were frozen by using 1.5M EG or 1.8M EG with/without 0.2M sucrose or O.1M trehalose. The development rate of blastocysts on day 7 after insemination in SOF+5% CS was significant higher than in TCM199 or CR1aa(P<0.05). The appearance rate of blastocysts on day 7-8 after insemination was higher than in TCM199, when fertilized oocytes were cultured in GRlas or SOF. The survival rate of frozen blastocysts after thawing tended to increase, when blastocysts were frozen by using 1.8M EG with 0.2M sucrose or O.1M trehalose. These results indicated that SOF or CRlaa media with amino acids was superior to TCM199 with monolayer in terms of blastocyst development in culturing of in vitro fertilized bovine nocytes, and sucrose or trehalose was supposed to prevent embryos from the freezing shock.

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Photocatalytic Cell Disruption of Giardia lamblia in a $UV/TiO_2$ Immobilized Optical-Fiber Reactor

  • 유미진;김병우
    • Journal of Microbiology and Biotechnology
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    • 제14권6호
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    • pp.1105-1113
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    • 2004
  • Disinfection of a waterborne pathogenic protozoa, Giardia lamblia, by the conventional chlorine method has been known to be difficult. An alternative disinfection has been carried out by using a UV -light illuminating optical­fiber photoreactor. Light intensity diffused from one piece of a clad-removed optical-fiber was $1- 1.5{\mu}Em^{-2}s^{-1}$. Disinfection capability in a UV -light irradiated optical-fiber reactor suspended with 0.01 g $TiO_{2}\;dm^{-3}$ was 1.4 times that in the same reactor without $TiO_{2}$ photocatalysts. To resolve the absorption and scattering of UV light by the particles themselves as well as the difficulty of recycling particles in the slurry­type reactor, $TiO_{2}$ which was obtained by a hydrothermal method, was immobilized on clad-removed optical fibers. Such pretreatment of fiber surface resulted in an excellent transparency, which enhanced the UV light to diffuse laterally from a fiber surface. Coating time of the prepared solution by the hydrothermal method was not effective after more than two times. Disinfection capability in the $TiO_{2}$-immobilized optical-fiber reactor was $83\%$ in 1 h at $40^{\circ}C$, which was slightly higher than $76\%$ at $22^{\circ}C$ and $68\%$ at $10^{\circ}C$. Disinfection capability at $22^{\circ}C$ increased from $74\%$ at an initial pH of 3.4, through $76\%$ at pH 6.5, to $87\%$ at an initial pH of 10. Oxygen supply with air-flow rate of 5 $cm^3\;min^{-1}$ did not seem to increase the disinfection capability with UV /immobilized $TiO_2$.

우 난포란의 체외성숙에 관한 연구 III. 체외성숙 우난포란의 체외수정과 발달 (Studies on In Vitro Maturation of Bovine Follicular Oocytes III. In Vitro Fertilization and Development of In Vitro Matured Bovine Follicular Oocytes)

  • 박세필;박태균;윤산현;고대환;정길생
    • 한국가축번식학회지
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    • 제13권2호
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    • pp.105-112
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    • 1989
  • These experiments were carried out to obtain the basic information for in vitro fertilization and development of bovine follicular oocytes matured in vitro. The bovine ovaries were obtained at a slaughter house and the follicular oocytes surrounded by cumulus cells were collected by puncturing follicles with 2-6mm of diameter. Bovine oocytes were matured in vitro for 24-26 hours in a CO2 incubator with 5% CO2 in air at 39$^{\circ}C$. The medium used for maturation was TCM199 supplemented with hormones, pyruvate, FBS and antibiotics. Epididymal spermatozoa were capacitated by in vitro culture for 2-3 hours in BO solution containing bovine serum albumin(5mg/ml) and caffein(2.5mM). Insemination was made by introducing about 10-15 matured oocytes into the suspension of capacitated spermatozoa. Six hour after inseminatin the eggs were transferred to TCM 199 supplemented with FBS(10%) for in vitro development. The results obtained in these experiments were summarized as follows : 1. The maturation rate of oocytes following incubation for 24-26 hours was 78.4%(228/291). 2. Of total 250 oocytes, 172 embryos extruded 2nd polar body following in vitro culture with spermatozoa for 20 hours, and the rates of embryos developed to 2-, 4-, 8-, 16-cells and morula or early blastocyst were 64.0, 39.2, 22.0, 15.2 and 11.2%, respectively. 3. The time needed for development to 2-, 4-. 8-, 16-cell stage and morula was 42.5$\pm$5.4, 58.0$\pm$9.2, 74.4$\pm$11.5, 96.1$\pm$13.4 and 119.0$\pm$18.2 hours, respectively.

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난구, 난관 상피세포 및 자궁 내막세포와의 공동배양이 돼지 난포란의 체외수정 및 분할율에 미치는 영향에 관한 연구 (Studies on the Effects of Co-Culture with Cumulus Cells, Oviduct Epithelial Cells and Uterine Endometrial Cells on in-vitro Fertilization and Cleavage Rate of Porcine Oocytes)

  • 김상근;김명헌;이무강;이규승
    • 한국가축번식학회지
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    • 제17권2호
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    • pp.133-139
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    • 1993
  • The studies were carried out to investigate the effects of co-culture with cumulus cell, oviduct epithelial cells and uterine endometrial cells on the in-vitro fertilization and cleavage rate of porcine follicular oocytes. The ovaries were obtained from slaughtered swine. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluids from the visible follicles of diameter 3~5 mm. The follicular oocytes were cultured in TCM-199 medium containing hormones and 10% FCS for 24~48 hrs in a incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation. The results obtained in these experiments were summarized as follows : 1. The in-vitro maturation and fertilization rate of porcine oocytes co-cultured with cumulus cells in TCM-199 meidum were 64.6%~74.5% and 37.5%~55.3%, respectively. And in-vitro fertilization rate of cumulus-enclosed oocytes(51.5%) were significantly(p<0.05) higher than cumulus-denuded oocytes(21.7%). 2. The in-vitro maturation and fertilization rate of porcine oocytes co-cultured with 1$\times$104 cells/ml, 1$\times$106 cells/ml, 1$\times$108 cells/ml and 1$\times$1015 cells/ml oviduct epithelial cells in TCM-199 medium were 53.5% and 37.2%, 61.7% and 46.8%, 54.5% and 31.8%, 42.2% and 26.7%, respectively. 3. The in-vintro maturation and fertilization rate of porcine oocytes co-cultured with 1$\times$106/ml, 1$\times$108/ml, 1$\times$1015/ml uterine endometrial cells in TCM-199 medium were 54.3% and 39.1%, 58.3% and 43.8%, 55.5% and 33.3%, and 45.7% and 30.4%, respectively. 4. When the in-vitro fertilized oocytes were co-cultured with porcine cumulus cells, ovdiduct epithelial cells and uterine endometrial cells, the development rate to the blastocyst stage was 9.5%, 10.7% and 11.8%, respectively and the rates were higher than that of control, 2.1%(p<0.05).

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토마토 공정묘의 상업적 생산을 위한 배지구성물질로서 발포유리와 탄화 밤나무 칩의 이용 (Use of CGF and CCW as Medium Components for Commercial Production of Plug Seedlings of Seogun Tomato)

  • 황승재;정병룡
    • 생물환경조절학회지
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    • 제13권2호
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    • pp.81-89
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    • 2004
  • 본 연구는 배지 구성물질로서 발포 유리질 소재(CGF)와 탄화 밤나무 목재입자(CCW)를 이용하여 배지를 재발하고 원예재배용 배지의 국산화를 위해 수행되었다. 토마토 종자는 발포 유리질 소재, 탄화 목재입자, 입상암면(GR), 그리고 피트모스를 혼합한 4종의 배지를 만들어 플러그 트레이에 충진하여 종자를 200구 플러그 트레이에 4반복으로 파종한 후 fog 번식상에서 3일간 발아시켰다. 발아 후 양지붕형 유리온실에 난괴법으로 배치하여 저면관수로 양액을 공급하여 육묘하였다. 상업적으로 이용되는 공정육묘 배지(토실이)를 대조구로 이용하였다. 플러그묘의 생육은 파종 후 31일째 측정하였다. 모든 처리에서 유사한 생육결과를 나타냈지만 특히 CCW(67%)+peatmoss(33%) 혼합배지에서 초장, 뿌리등급, 생체중, 그리고 배지의 기상과 가비중이 다른 처리에서보다 유의성 있게 좋았다. 모든 배지의 pH는 6.3∼7.4의 범위로 측정되었다. CGF와 입상암면의 혼합비율이 증가할수록 배지의 pH는 증가하였다. 재배 후 배지의 EC는 대조구에서 가장 높았으나 독성증상은 관찰되지 않았다. 엽수와 엽록소 함량은 처리간 유의성이 없었다.

Effects of alkali solutions on corrosion durability of geopolymer concrete

  • Shaikh, Faiz U.A.
    • Advances in concrete construction
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    • 제2권2호
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    • pp.109-123
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    • 2014
  • This paper presents chloride induced corrosion durability of reinforcing steel in geopolymer concretes containing different contents of sodium silicate ($Na_2SiO_3$) and molarities of NaOH solutions. Seven series of mixes are considered in this study. The first series is ordinary Portland cement (OPC) concrete and is considered as the control mix. The rest six series are geopolymer concretes containing 14 and 16 molar NaOH and $Na_2SiO_3$ to NaOH ratios of 2.5, 3.0 and 3.5. In each series three lollypop specimens of 100 mm in diameter and 200 mm in length, each having one 12 mm diameter steel bar are considered for chloride induced corrosion study. The specimens are subjected to cyclic wetting and drying regime for two months. In wet cycle the specimens are immersed in water containing 3.5% (by wt.) NaCl salt for 4 days, while in dry cycle the specimens are placed in open air for three days. The corrosion activity is monitored by measuring the copper/copper sulphate ($Cu/CuSO_4$) half-cell potential according to ASTM C-876. The chloride penetration depth and sorptivity of all seven concretes are also measured. Results show that the geopolymer concretes exhibited better corrosion resistance than OPC concrete. The higher the amount of $Na_2SiO_3$ and higher the concentration of NaOH solutions the better the corrosion resistance of geopolymer concrete is. Similar behaviour is also observed in sorptivity and chloride penetration depth measurements. Generally, the geopolymer concretes exhibited lower sorptivity and chloride penetration depth than that of OPC concrete. Correlation between the sorptivity and the chloride penetration of geopolymer concretes is established. Correlations are also established between 28 days compressive strength and sorptivity and between 28 days compressive strength and chloride penetration of geopolymer concretes.

Babesia gibsoni의 적혈구내 배양법과 진단법 개발에 관한 연구 2. Babesia gibsoni의 적혈구내 배양 (Intraerythrocytic culture and development of serological diagnostic tests of Babesia gibsoni 2. Intraeryrhrocytic culture of Babesia gibsoni by microaerophilous stationary phase(MASP))

  • 서명득;주보현
    • 대한수의학회지
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    • 제38권2호
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    • pp.359-365
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    • 1998
  • This study was conducted to isolate the protozoan parasite Babesia gibsoni by intraerythrocytic culture method of micoraerophilous stationary phase(MASP) and evaluate the possibility of application for the detection of B gibsoni in canine babesiosis. Also, indirect fluorescent antibody test(IFAT) and thick blood smear(giemsa stain), direct light microscopy (DLM), as control diagnostic tests, were conducted to compare diagnostic effects between MASP, IFAT and DLM. The results obtained from this study were summarized as follows. The protozoan parasite B gibsoni multiplied in 24-well polystyrene plate containing 1.2ml of canine red blood cell suspension in RPMI 1640 medium(pH 7.0) which is contained 20~40% normal canine serum(NCS) under the MASP condition of 5% $CO_2$ and 95% air at $37^{\circ}C$ incubator. Under the above MASP culturing system the percentage of parasitized erythrocytes(PPE) after incubation for 9 days reached the peak. The levels of PPE in MASP culture were shown more higher by exchanging the medium at 24 hour intervals. The parasite were purely isolated from MASP culture of canine red blood cells collected from dogs(pit bullterrier) infected with B gibsoni naturally. Among the total of 83 heads of pit bullterrier blood samples the positive rate was 32 heads(38.5%) in DLM, 45 heads(54.2%) in IFAT and 42 heads(50.6) in MASP culture. In negative cases of IFAT and DLM the isolation rates of B gibsoni by MASP culture were 16 heads(42.1%) of 38 heads and 16 heads(28.6%)% of 56 heads, respectively. From this study it was suggested that MASP culture method by RPMI 1640 medium was a reliable and useful diagnostic test for the diagnosis of B gibsoni infections in canine babesiosis.

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