• Journal of Animal Science and Technology
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• v.55 no.5
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• pp.417-425
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• 2013

We sought to provide a method for freezing and preserving primordial germ cells, or an avian germ cell of a bird, as a material for developmental engineering or species preservation. The aim of this study was to compare the efficacy of slow freezing with a vitrification method for the cryopreservation of chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonad of day 5.5-6 day (stage 28) cultured chick embryos, using the MACS method, were classified into two groups: slow freezing and vitrification. We examined the viability of PGCs after Cryopreservation. Four freezing methods were compared with each other, including the following: Method 1: The PGCs were frozen by a programmed freezer in a plastic straw, including 2.0 M ethylene glycol (EG) as cryoprotective additive (slow freezing) Method 2: The PGCs were vitrified in a plastic straw, including 8.0 M EG, plus 7% polyvinylpyrrolidone (PVP) (rapid freezing). Method 3: The slow freezing was induced with a cryotube including 2.0 M EG Method 4: The PGCs were frozen in a cryotube including 10% dimethyl suloxide (DMSO) (rapid freezing). After freezing and thawing, survival rates of the frozen-thawed PGCs from Method 1 to 4were 76.4%, 70.6%, 80.5% and 78.1% (p<0.05), respectively. The slow freezing ($-80^{\circ}C$ programmed freezer) method may provide better survival rates of frozen-thawed PGCs than the vitrification method for the cryopreservation of PGCs. Therefore, these systems may contribute to the cryopreservation of a rare avian species.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.10 no.1
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• pp.23-29
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• 1977

Three kinds of samples, Trachpenalus curiviostris, Astroconger myriaster and Cantherines modestus which were pre-treated in a processing plant were frozen at $-40^{\circ}C$ in a contact freezer and stored for 32 days.The numbers of general bacterium, coliforms and E. coli were measured at 8 day intevals during frozen storage and the isolated strains was classified. The results are as follows; 1. The numbers of coliforms and E. coli in the samples before freezing were much higher, than those during frozen storage and it tended to decrease. 2. General bacteria showed little change in number before and after being frozen. Among 97 strains of isolated coliforms, only 4 strains of K. aerogenes I ana 4 strains of K, cloacae were classified and the rest was not determined. 3. Ninety percent of coliforms was found to be psychrotrophic coliforms, which were able to grow at $5^{\circ}C$ within a week. 4. Vibrio and Pseudomonas were superior in number before freezing while Flavobacterium cytophaga and Moraxella were superior during frozem storage.

• Journal of Sericultural and Entomological Science
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• v.36 no.1
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• pp.62-68
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• 1994

Isolation, indentification, bioassay and distribution of ice nucleation active(INA) bacteria were done on branch rot of mulberry which was severely developed after harvest of mulberry branches in autumn. Twelve isolates and two isolates out of thirty-six isolates had ice nucleation activity from -5$^{\circ}C$ to -1$0^{\circ}C$, and over -5$^{\circ}C$, respercively. Isolates which formed ice nucleation from -5$^{\circ}C$ to -1$0^{\circ}C$ were not inclined to injure tomato and corn seedlings. However, Two isolates, SE9316 and SE9338 which formed ice nucleation over -5$^{\circ}C$ injured mulberry, tomato and corn seedlings. SE9316 and SE9338 were identified as Pseudomonas syringae based on the morphological, cultural and biochemical characteristics. Populations of ice nucleation active bacteria, fluorecent pseudomonads, were higer in February and April, but they decreased in May. Populations decreased as they become distant from the center of the symptom. The bacterial populations of all sampling times and sites were higer than 105 cfu/g which was enough to induce frost injury.

• Microbiology and Biotechnology Letters
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• v.26 no.3
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• pp.200-205
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• 1998

A probiotic-strain of Lactobacillus sp. was cultured in bovine blood plasma-based (BBPB) medium and freeze-dried to prepare a probiotic product as an animal feed additive. The cell mass produced in the medium, $5.2{\times}10^9$ CFU/ml, was high enough to be commercialized and was 74% of that in MRS medium. The survival rate of tactobacillus sp. against freeze-drying was affected by the conditions for treatment of cultured BBPB broth before freeze-drying such as pH adjustment, volume reduction and freezing rate. It was also found that the blood protein hydrolysate remaining in broth also enhanced the survival rate. Among various protective substances, sucrose showed a high stabilizing effect with 10% (w/v) addition, by which the maximum survival rate (48.3%) and viable cell count ($3.0{\times}10^{10}$ CFU/g) were obtained.

• Journal of Embryo Transfer
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• v.21 no.1
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• pp.77-83
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• 2006

본 연구는 개 동결 정액 융해 시 straw 크기 및 융해 속도가 융해 정자의 질(quality)에 미치는 영향을 조사하고 최적의 융해 조건을 조사하는데 그 목적이 있다. 정상적인 번식능을 가진 비글 수컷 5마리에서 정액을 채취하여 원심 분리하여 정장을 버리고 남은 정자에 동결보호제인 glycerol이 첨가된 tris-glucose-egg yolk extender를 첨가하여 동결하고 액체질소에 보관한 후 융해하였다. 동결 융해 조건에 따른 효과를 알아보기 위해 straw는 0.25 ml과 0.5 ml크기를 사용하였고 융해 조건은 $75^{\circ}C$에 10초, $55^{\circ}C$에 12초 및 $37^{\circ}C$에서 120초로 하여 융해 후 정자의 활력도(vigor), 운동성(motility), Hypo-osmotic test(HOS test)를 이용한 생존성(viability) 및 $SperMac^{\circledR}$ 염색을 하여 정자의 membrane integrity를 비교 조사하였다. 조사 결과 0.5 ml 크기의 straw를 사용한 경우 $37^{\circ}C$ 융해가 $55^{\circ}C,\;75^{\circ}C$ 융해보다, 0.25 ml 크기의 straw를 사용한 경우에는 $37^{\circ}C,\;55^{\circ}C$ 융해가 $75^{\circ}C$ 융해보다 유의적으로 높은 활력 지수 및 생존성을 보였다(P<0.05). Straw크기에 따라 비교하였을 경우 0.5 ml 군에서 유의적으로 높은 활력도, 생존성 및 membrane integrity를 보였다(P<0.05). 결론적으로 개 정액이 동결 및 융해 시 0.5ml straw를 이용하여 동결한 후 $37^{\circ}C$에서 120초 동안 융해하는 것이 최적의 조건임이 사료된다.

• 한국신재생에너지학회:학술대회논문집
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• 2007.06a
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• pp.212-215
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• 2007

고분자전해질연료전지 시스템이 영하 조건에 노출될 경우, 셀의 성능 및 내구성에 미치는 영향을 실험적으로 확인해 보았다. -30 $^{\circ}C{\sim}70$ $^{\circ}C$ 조건을 반복 경험시키며, 성능저하 정도를 살펴보았다. 일반적인 운전조건과 동결/해동에 의한 성능저하 요인을 분리하여 확인하기 위해, 30 $^{\circ}C{\sim}70$ $^{\circ}C$ 범위의 사이클을 진행한 경우에 대해, 위와 통일한 분석을 통하여 성능 및 각종 물성 값의 변화를 비교하였다. 동결조건에서 셀의 성능저하는 형성된 얼음의 물리적 부피팽창으로 인한 계면저항의 증가가 주요 원인임을 밝힐 수 있었다.

• Korean journal of food and cookery science
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• v.12 no.3
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• pp.312-319
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• 1996

Oyster (Crassostrea virginica) and Weakfish (Cynoscion regalis) were stored at 6, 0, -4 and -20$^{\circ}C$ for up to 45 days and examined for changes in microflora. Aerobic plate counts (incubated at 21$^{\circ}C$) were performed at selected times during storage and 495 isolates (255 isolates from oyster and 240 isolates from Weakfish) were randomly selected from the plates during the storage. Before the storage of the fishes, viable counts of oyster were 4.9${\times}$10$\^$5/ CFU/g of meat and those of Weakfish were 1.5${\times}$10$^4$ CFU/cm$^2$of skin. Microflora of oyster before storage, the major isolates identified as Pseudomonas spp. (67%) and Vibrio spp. (20%). Pseudomonas ll1/1V-H and Flavobacterium/Cytophaga were predominant genus in the microflora of oyster during cold storage at 6, 0, -4 and -20$^{\circ}C$. The composition of the microflora of Weakfish before storage, Acinetobacter (40%) and Moraxella (33%) were the major species, with Pseudomonas and Vibrio constituting a small percentage of the total isolates. The microflora shifted to predominantly Pseudomonas spp. during storage at 6. 0 and -4$^{\circ}C$, making up from 60 to 100％ of isolated strains. During frozen storage, the percentage of isolates identified as Mnraxella increased to 40-60% of the total isolates. During cold storage, halophilic bacteria (Pseudomonas lII/IV-H and Vibrio) were predominant in oyster while nonhalophilic bacteria (Pseudomonas III/IV-NH and Moraxella) were predominant in Weakfish. Vibrio spp. were higher in oyster than in Weak fish. Listeria spp. were not isolated but unidentified ${\beta}$-hemolytic bacteria were islolated from both of the fishes during cold storage.

• Korean Journal of Poultry Science
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• v.40 no.3
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• pp.163-169
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• 2013

Cryopreservation of poultry semen has been reported, but preservation of female genetic material has not been possible because of the unique anatomical and physiological characteristics of the avian egg. Thus an alternative strategy for conservation of oviparous species of animals must be developed. Recent technological developments for producing germline chimeras by the transfer of primordial germ cells (PGCs) into recipient embryos has enabled the conservation and retrieval of chicken genetic resources in their complete form. In the present study, fertilized eggs were incubated for about 5.5 days to obtain embryos at stage 28. The whole embryo was collected from the germinal gonad using a fine glass micro pipette under a microscope. The PGCs were then purified using MACS method. Two commercially available cryoprotectants (A and B) were used to preserve the PGCs, and EG were used as a control. The average recovery rate of PGCs after thawing was 35.5% and 60.5% with the A and B treatments, respectively. There was no significant difference between B treatments and control, which showed an average recovery rate of 52.8%. However, the recovery rate obtained using A cryoprotectant (35.5%) was significantly lower than using treatment control and B. The average viability of the PGCs after thawing were 77.9% and 77.4% for cryoprotectants A and B, respectively, and the control were was 81.6%. There was no statistically significant difference between the two treatments and control. It was concluded that all of the available cryoprotectants examined in this study could be used for preservation of PGCs from embryos. Further experiments to produce germline chimera from PGCs preserved using this techniques are strongly recommended.

• Korean Journal of Food Science and Technology
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• v.31 no.6
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• pp.1596-1603
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• 1999

Dongchimi (Korean-style fermented radish with juice) products were frozen to prevent further acidification and softening of texture by restraining microbial growth and enzyme activity during storage. Dongchimi juice and radish were separated prior to freezing process. Dongchimi radish was frozen at $-20^{\circ}C,\;-70^{\circ}C$ and immersed in liquid nitrogen and dongchimi juice was frozen at $-20^{\circ}C\;and\;-70^{\circ}C$. Frozen dongchimi samples were thawed with ambient temperatures of $4^{\circ}C\;and\;27^{\circ}C$ and with 915 MHz-microwave, respectively. Dongchimi radish immersed in liquid nitrogen and thawed with 915 MHz-microwave showed the highest pectinesterase activity and hardness, and the lowest polygalacturonase activity and color change, indicating that this quick freezing-quick thawing method can be used for the long-term storage of dongchimi products. Dongchimi juice frozen at $-70^{\circ}C$ and thawed with 915 MHz-microwave retained its pH and titrable acidity, and showed a largest reduction in total aerobic count and lactic acid bacteria.

• Korean Journal of Food Science and Technology
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• v.22 no.1
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• pp.44-49
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• 1990

The quality of garlic powder produced by hot all drying and freeze drying method was evaluated Porapak Q column was comparatively profitable for the separation of flavor components in hexane extracts of garlic. Freeze dried (FD) garlic powder had higher diallyl disulphide, total pyruvic acid and alliin than hot air dried (HD) garlic powder. On the results of color evaluation by color difference meter, garlic powder of HD were more brownish than FD. In microstructure of garlic powder observed by SEM, freeze dried garlic powder was fairly porous.