• 제목/요약/키워드: thawing conditions

검색결과 152건 처리시간 0.025초

Effect of frozen storage and various concentrations of sucrose media on survivability of enterotoxigenic Escherichia coli (ETEC) for oral challenge of weaner pigs

  • Cho, Hyun Min;Kang, Joo Won;Kim, Yeong Kuk;Lee, Joo Bin;Oh, Chan Yi;Heo, Jung Min;Yi, Young-Joo
    • 농업과학연구
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    • 제43권5호
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    • pp.788-793
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    • 2016
  • Post-weaning diarrhea (PWD), mostly caused by enterotoxigenic Escherichia coli (ETEC), remains to be a major source of economic loss in swine industry. The use of the ETEC-oral challenge model is often applied to mimic unsanitary commercial swine farm conditions where pathogens and unknown complex microbes exist and can cause severe infections in pigs. The purpose of this study was (1) to estimate ETEC density using spectrophotometric computation, (2) to determine survivability of ETEC after storing at $-20^{\circ}C$ for 7 days, and (3) to evaluate survivability of ETEC after blending with diluted sweeteners (0, 5, 10, 20, and 40% sucrose in phosphate buffered saline [PBS]). Cell density was quantified using UV-VIS spectrophotometer and counting ETEC colony forming units (cfu) at 0, 30, 60, 90, 120, 150, 180, 210, and 240 min. The established linear equation ($y=0.0031x^2-0.0079x+0.0043$ and $y=0.0046x^2-0.0151x+0.0113$) was used for robust quantification of each ETEC cell density. ETEC stored at $-20^{\circ}C$ showed 108 cfu/mL after thawing and incubation. When ETEC was blended with sweeteners (20 and 40%), survival of ETEC was decreased by 58 and 54% in 5 min post blending. However, addition of 20% of sweetener resulted in a higher survivability than those with other media concentrations. Therefore, the use of ETEC-oral challenge model would be possible as a stable method if we could confirm the appropriate medium that increases survivability of ETEC in weaner pigs.

BTS와 Androhep이 보존 기간 동안 액상 정액의 운동역학 및 수정능 획득에 미치는 영향 (Effect of BTS and Androhep during Storage Times on the Kinematics and Capacitation Status in Liquid Boar Semen)

  • 김연희;박유진;윤성재;권우성;김상현;방명걸
    • Reproductive and Developmental Biology
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    • 제34권3호
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    • pp.241-246
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    • 2010
  • The objective of this study was to investigate the effect of storage time on fresh boar semen in Androhep and Beltsville Thawing Solution (BTS). Boar semen samples extended in each extender were stored at $17^{\circ}C$ up to 4 days. Sperm motility kinematics was evaluated by computer assisted sperm analyzer (CASA) and capacitation status by chlortetracycline (CTC)/Hoechst 33258 staining. Sperm motility (%) was not decreased during storage in BTS and Androhep. No significant difference between extenders was observed. Only significant differences in kinematic parameters on linearity during storage were found. The percentage of dead sperm significantly decreased during storage (p<0.05). Also the percentage of noncapacitated, capacitated, and acrosome-reacted sperm significantly modified during storage (p<0.05). However, there was no significant difference between extenders except proportion of capacitated sperm. This finding supported that modification in these parameters was not significantly different between extenders during this short-term storage. Our finding strongly indicated that both Androhep and BIS maintained favorable conditions for motility, motility kinematics, and capacitation status during short-term storage. Despite modifications in some parameters were apparent during sperm storage in extenders, these may not affect the fertilizing capacity of boar semen.

Competitive Enzyme-Linked Immunosorbent Assay for Glucose-6-Phosphate Dehydrogenase

  • Kim, Moon-Hee
    • BMB Reports
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    • 제30권5호
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    • pp.326-331
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    • 1997
  • To construct a competitive ELISA standard curve for the detection of glucose-6-phosphate debydrogenase (G6PD), we used highly purified native G6PD (nG6PD) as both immobilized and soluble antigens and anti-G6PD serum raised against nG6PD as antibody. The polystyrene cuvettes coated with nG6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of nG6PD as competitors followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA did not exhibit the typical sigmoidal dose-response curve characteristic of competition immunoassays under the optimal concentrations of antigen and antibody. The soluble nG6PD used as competitor failed to effectively inhibit the binding of antibodies to the immobilized nG6PD. The addition of NADP, a cofactor of G6PD enzyme, to coating buffer used for immobilizing nG6PD to the cuvettes and PBS-Tween-BSA buffer for diluting competitors did not improve the inhibition of antibody binding to immobilized nG6PD by soluble n/G6PD. The addition of BSA to coating buffer did not increase inhibition, either. Surprisingly, when partially active G6PD (paG6PD), obtained by repeated freeze-thawing, was used as competitor, the antibody binding to either immobilized nG6PD or immobilized paG6PD was inhibited 49-58%. We conclude that an effective competitive ELISA system with nG6PD enzyme and anti-G6PD serum for the detection of G6PD may not be established due to the poor inhibition of antibody binding to immobilized nG6PD by soluble nG6PD under the present assay conditions and that the inhibition may be improved by using an inactivated enzyme as competitor regardless of the type of immobilized antigen used. These results imply that the immobilized nG6PD may undergo denaturation upon binding to the polystyrene cuvettes and that our anti-G6PD serum may recognize denatured enzyme better than active enzyme.

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고성능유동화제가 콘크리트의 공학적 특성에 미치는 영향 (I) (A Study on the Effects of Superplasticizers on the Engineering Preperties of Plain Concrete (I))

  • 박승범;이보성
    • 한국농공학회지
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    • 제26권3호
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    • pp.68-80
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    • 1984
  • This study was attempted in order to investigate the effects of superplasticizers on fresh and hardened concrete. The experimental program included tests on the slump and slump loss, bleeding, time of set, air content, the compacting, factor Vee Dee, compressive strength, tensile and flexural strength, permeability, shrinkage and freege-thaw durability. The major conclusions that can be drawn on the study are as follows. 1. Superplasticizers were observed to have an appreciable fluidifying action in fresh concrete so that tinder appropriate conditions, they either considerably improved its workability or permitted a water reduction of at least 8-12% to be made while maintaining normal workability. 2. The bleeding ratios of base and S,P. Concrete were much lower than those of the conventional concrete. Differences between the base and S.P. Concrete were insignifician. 3. The setting time was the longest for conventionla concrete, followed by S.P. concrete and base concrete in thatorder. And AE water reduction admixtures showed an appreciable influence on the setting and hardening characteristics of concrete and prolonged the stiffening times. 4. The high initial slump values of S.P. concrete generally decreased rapidly with increased standing time. CF values showed increasing tendencies with the increase of S.P. content, and excessive addition of S.P. caused the segregation of fresh concrete, resulting in its rejection. 5. Though there was a slight increase in strength, no significant differences are observed between base and S.P. concrete in terms of the compressive, tensile and flexural strength. 6. The permeability of S.P. concrete was significantly less than that of the conventional concrete, and the shrinkage of S.P. concrete was considerably smaller than that of the conventional concrete, but there were no significant differences between base and S.P. concrete. 7. Compared to base concrete, S.P. concrete without entrained air tended to slightly increase freeze-thaw durability, and S.P. concrete with an appropriate entrained air gave satisfactory resistance to freezing and thawing.

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Establishment and long-term culture of the cell lines derived from gonad tissues of Siberian sturgeon (Acipenser baerii)

  • Ryu, Jun Hyung;Nam, Yoon Kwon;Gong, Seung Pyo
    • Fisheries and Aquatic Sciences
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    • 제19권4호
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    • pp.16.1-16.8
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    • 2016
  • To culture germline stem cells in vitro, establishment of the cell lines that can be used as the feeder cells is a prerequisite. In this study, we tried to establish gonad-derived cell lines in Siberian sturgeon (Acipenser baerii). Five 1-year-old A. baerii were used as a donor of gonad tissues, and gonad-dissociated cells were cultured in vitro. Subsequently, determination of growth conditions, long-term culture, characterization, and cryopreservation of the cell lines were also conducted. Five gonad-derived cell lines were stably established and cultured continuously over at least the 73th passage and 402 culture days under the media containing 20 % fetal bovine serum at $28^{\circ}C$. All cell lines consisted of two main cell types based on morphology even if the ratio of the two cell types was different depending on cell lines. Despite long-term culture, all cell lines maintained diploid DNA contents and expression of several genes that are known to express in the A. baerii gonad. After freezing and thawing of the cell lines, post-thaw cell viabilities between 57.6 and 92.9 % depending on cell lines were indentified, suggesting that stable cryopreservation is possible. The results and the cell lines established in this study will contribute to the development of an in vitro system for A. baerii germline stem cell culture.

복합양생에 의한 폴리머 시멘트 모르타르의 성질 (Properties of Polymer Cement Mortars under Combined Cures)

  • 조영국
    • 콘크리트학회논문집
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    • 제18권5호
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    • pp.667-675
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    • 2006
  • 대분분의 콘크리트 구조물은 여러 가지 환경요인에 노출되었는데, 여러 가지 환경요인 중, 요인 한가지만의 작용이 아니라 여러 가지 요인이 복합적으로 작용하여 콘크리트의 내구성은 현격하게 저하하게 된다. 본 연구에서는 복합양생을 받는 폴리머 시멘트 모르타르에 관한 강도 및 내구성 개선효과를 실험하였다. 폴리머 종류, 폴리머 시멘트비, 시멘트-잔골재비를 변화하여 폴리머 시멘트 모르타르를 제작한 후, 5가지의 환경요인, 즉 양생조건을 달리하여, 강도성상, 중성화, 염화물이온 침투, 내동해성, 내산성을 시험하였다. 시험결과, 폴리머 시멘트 모르타르는 보통 시멘트 모르타르에 비해 우수한 강도성상을 나타냈으며, 양생조건 II와 V에서 강도가 개선되었으나, 양생조건 I와 IV에서는 강도의 발현이 낮았다. 또한 중성화 및 염화물이온 침투 깊이는 양생조건 III-C, IV-B, V-A 순으로 작아졌으며, 폴리머 시멘트비의 증가에 따라 감소하였다. 본 연구에서 폴리머 시멘트비의 적정 혼입비는 10% 또는 15%이었다.

생쥐배아의 냉동보존에 있어서 여러 조건의 평가 - 저속 처리단계와 급속 처리단계, 배양액, 세포기 (The Evaluation of Various Conditions in the Cryopreservation of Mouse Embryos - Rapid and Slow Method of Cryopreservation, Culture Media and Cell Stages)

  • 이승연;권주택;송희원;조윤희;이기숙;류철희;김종덕
    • Clinical and Experimental Reproductive Medicine
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    • 제26권2호
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    • pp.127-135
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    • 1999
  • Cryopreservation is able to store the surplus pre-embryos for freezing and furthermore thawing and transfer in a subsequent cycle. Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps, freezing media and embryonic stages on the rates of viability and development of cryopreserved mouse embryos. Female ICR mice ($6{\sim}8$ weeks old) were induced to superovulate by sequential intraperitoneal injection of 5 IU PMSG and 5 IU hCG 48h apart. Mouse embryos were collected according to its developmental stage after the injection of hCG. Embryos were cryopreserved not only by cryoprotectant step (1 step${\sim}$4 step) but also in a variety of media (HTF, IVF medium, D-PBS) and cell stage. The results were as follows: There is no clear advantage in these freezing media of rapid method, but 4 cell and 8 cell of slow method (2, 3, 4 step) have advantage in D-PBS. The development of embryos according to cell stage become greater in 8 cell stage. In the treatment steps of cryopreservation, the development of embryo to blastocyst was similar among rapid method, but the development of 4 cell and 8 cell embryos to blastocyst according to slow method was better than rapid method.

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다층구조확산을 고려한 제설제에 노출된 콘크리트의 염화물 해석 (Analysis for Chloride Penetration in Concrete under Deicing Agent using Multi Layer Diffusion)

  • 서지석;권성준
    • 한국콘텐츠학회논문지
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    • 제16권4호
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    • pp.114-122
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    • 2016
  • 콘크리트는 경제적이며 고내구성 건설재료이지만, 염해에 노출된 경우 매립된 철근의 부식으로 인해 내구성에 대한 문제가 발생한다. 최근 들어 동절기에 제설제가 많이 사용되고 있는데, 제설제의 사용은 콘크리트 표면에 미세균열과 박리를 증가시키고 용해된 제설제는 매립된 철근의 부식을 야기한다. 기존의 염화물 지배방정식인 Fick's 2nd Law의 해석기법은 표면이 열화된 콘크리트의 염해특성을 평가하지 못하므로 이에 대한 고려가 필요하다. 본 연구에서는 콘크리트 다층구조 확산 모델과 시간의존성 염화물 확산을 이용하여 제설제에 노출된 콘크리트의 염화물 해석기법을 제안하였다. 이를 위해 18년 경과된 콘크리트 도로교의 염해실태를 분석하였으며, 역해석을 통하여 표면열화깊이 및 열화된 콘크리트 층의 증가된 염화물 확산성을 평가하였다. 제안된 기법은 30MPa 콘크리트에서 12.5~15.0mm 열화깊이와 2배 증가된 열화층의 염화물 확산성을 나타내었다. 본 해석기법은 표면열화 및 표면 강화 등 2개의 다른 확산성을 가진 콘크리트의 염화물 거동을 평가하는데 효과적으로 적용될 수 있다.

고강도(高强度) 콘크리트에서 기포조직(氣泡組織) 및 강도특성(强度特性)이 연결융해저항(連結融解抵抗)에 미치는 영향(影響) (Effect of Air Void and Strength Characteristics with Freezing and Thawing Resistance on High Strength Concrete)

  • 김생빈;문제길;김동신
    • 대한토목학회논문집
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    • 제13권3호
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    • pp.13-24
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    • 1993
  • 본(本) 연구(硏究)는 고강도(高强度) 콘크리트가 외부(外部)로부터의 수분(水分)의 공급(供給)을 받으면서 상온(常溫)에서 저온(低溫)까지의 가혹한 기상작용을 되풀이 받을 때 고강도(高强度) 콘크리트의 연결융해(連結融解)저항에 미치는 기포조직(氣泡組織) 및 강도특성(强度特性)의 영향에 대한 실험적(實驗的) 연구(硏究)이다. 이 연구(硏究)를 위하여 1단계 실험(實驗)에서는 보통 포틀랜드 시멘트를 사용한 AE콘크리트와 non-AE 콘크리트에 대해 물 시멘트비와 공기량을 중심(中心)으로 내연해성(耐連害性)과 기포조직과의 관계를 규명하고, 2단계 실험(實驗)에서는 non-AE 콘크리트의 내연해성(耐連害性)을 향상(向上)시키기 위해 공시체(供試體) 제작시 진동다짐으로 세심한 주의를 기울이고, 또 동결융해 시험시의 초기강도를 높이기 위해 조강(早强)포틀랜드 시멘트를 사용하여 고수밀성(高水密性)의 콘크리트를 제작하여 실험하였다. 이때 수밀성(水密性)를 투수시험(透水試驗)에 의한 투수계수(透水係數)를 측정하여 판단하고 아울러 고수밀성(高水密性) 콘크리트의 기포조직(氣泡組織)도 측정하였다.

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복어(Lagocephalus wheeleri) 수리미를 첨가한 어묵의 제조 및 품질특성 (Processing and Quality of Natural-tasting Steamed Fish Paste Containing Unwashed Pufferfish Lagocephalus wheeleri Surimi)

  • 안병수;김병균;황석민;박노현;이현진;오광수
    • 한국수산과학회지
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    • 제52권6호
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    • pp.562-570
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    • 2019
  • To develop natural-tasting fish paste from the pufferfish Lagocephalus wheeleri, steamed pufferfish paste (SPP) was prepared and its optimal processing conditions, quality metrics, and shelf-life characteristics were examined. SPP was produced by thawing golden threadfin Nemipterus virgatus surimi (FA grade), then adding 10% unwashed pufferfish surimi (PS), 1.5% salt, 8.0% wheat starch, 0.25% calcium carbonate, 0.25% sugar, 0.75% sorbitol, 0.25% polyphosphate, and 12.0% pufferfish hot-water extract (Brix 10°). The meat was ground with a Stephan mixer, molded at low temperature (18℃, 10 h), vacuum packed in a laminated plastic film bag, heat treated with hot water (95℃, 50 min), and cooled. As the amount of PS added increased, the whiteness, gel strength, and shear strength of the SPP decreased slightly. However, the SPP folding test showed no deterioration in the texture. In the sensory evaluation, the SPP received a higher rating for taste, smell, and overall taste than commercial Japanese pufferfish Kamaboko. The total amino acid content of the SPP was 10,262.6 mg/100 g; the major amino acids were aspartic acid, glutamic acid, alanine, valine, leucine, lysine, and arginine. The free amino acid content was 133.0 mg/100 g; the major amino acids were taurine, glutamic acid, glycine, alanine, cystine, and lysine.