This study was conducted to establish a freezing method of miniature pig spermatozoa. The semen ejaculated from PWG M-type miniature pig was collected by gloved-hand method. The semen was diluted with same volume extender (m-Modena B). The frozen solution used frozen solution of four different (LEY, TCG, BF-5 and m-Modena+egg yolk) for find optimal frozen solution in miniature pig sperm. The diluted semen for frozen rate assay was added to LEY solution (solution Ⅰ: 11% lactose+egg yolk; solution Ⅱ: solutionⅠ+glycerol+OEP), and frozen depending on freezing rate by the three different freezing methods (A: until 5℃ for 1 hrs, holding at —102℃ for 10 min; B: until 5℃ for 2 hrs, holding at —102℃ for 10 min; C: until 5℃ for 3 hrs, holding at —80 and —102℃ for 10 min). Semen cooled until 5℃ was added with glycerol 1, 3 and 5%, and take a equilibrium time for 0, 10 and 30min. Frozen-thawed sperm were evaluated for viability, acrosomal status and morphological abnormality. The results of frozen-thawed sperm ability by frozen solution, viability was higher in LEY solution compared to other three different frozen solution. AR pattern of LEY solution were lower than other three different frozen solution. The results of freezing rate, viability was higher in B method compared to other methods (p<0.05). Acrosomal statute was intacted in A and B methods than C method. The experiment for glycerol condition was showed that sperm viability was higher in extender with 1% and 3% glycerol and equilibrium time of 0 min. The acrosome damage was lower in extender with 1% glycerol and equilibrium time of 10 min than other conditions. In conclusion, the optimal conditions for cryopreservation of miniature pig spermatozoa obtained in LEY frozen solution, cooling rate of 1～2 hrs, 1～3% glycerol concentrations and glycerol equilibrium time of 0～10 min.

This study was undertaken to find out the effect of methyl-beta-cyclodextrin (MBCD) on cold shock and membrane cholesterol quantity of sperm during the freezing process in miniature pigs. For this study, semen ejaculated from PWG M-type miniature pig was diluted that freezing solution (with egg yolk group) and m-Modena B (without egg yolk group) treated with 0, 1, 5, 10 and 20 mM MBCD before freezing process. The diluted semen was monitored sperm ability at room temperature, after cooled until 5℃ and after forzen-thawed for cold shock test of spermatozoa. Also, membrane cholesterol of sperm was extracted by folch solution at the same time. sperm ability was assessed for viability and acrosomal status. The membrane cholesterol quantity was measured by thin-layer chromatography (TLC) method. The result, viability and acrosome integrity in semen diluted without egg yolk groups were decreased at all temperature range by increasing of MBCD concentration. In particular, sperm of egg yolk group was showed that significantly higher viability and lower acrosome damage when treated with 5 mM MBCD (p<0.05). The results of TLC experiment, cholesterol amounts were increased with MBCD cocentration in egg yolk, and decreased with MBCD concentration in m-Modena B. In cryopreservation efficiency, there was no significant difference at viability, and acrosomal state of sperm in 5 mM MBCD concentration was significantly lower in acrosome damage than other groups (p<0.05). Therefore, the addition MBCD in egg yolk was protected spermatozoa from cold shock injury. This protective effect of MBCD may be due to addition of sperm membrane cholesterol.

Arcobacter butzleri is one of the aerotolerant Campylobacter species which cause persistent diarrhea,abdominal pain, nausea and vomiting in human. The aim of this study was to determine the growth and survival of A. butzleri under the heat treatment or freezing storage condition. In heat treatment, two Korean isolates of A. butzleri were treated at 40 to 80℃ for various times. In freeze treatment, two Korean isolates of A. butzleri were kept at 4, −20 and −70℃ from 1 to 15 d. The survivability of the A. butzleri Korean isolates significantly decreased at higher than 60℃ heating condition but it didn't show any significant difference in 40℃ treated group. Under the cold stress condition, survivability of A. butzleri were significantly decreased at −20℃ storage. Like other foodborne pathogens,survivability of A. butzleri was controlled by heat and freezing treatment.

최근 전통 식품에 대한 관심이 증가함에 따라서 우리 나라 고유의 술인 막걸리 소비가 국내에서뿐만 아니라 해외 수출량도 급증하고 있다. 그러나 막걸리의 유통 및 저장 안정성을 증가시킬 수 있는 방법으로는 미생물 제어 기술로서 냉장 유통과 가온 살균에 제한되어 표준화된 기술이 부족한 실정이다. 따라서 본 연구에서는 막걸리 유통기간 연장 방법의 하나로 막걸리를 급속동결(Quick Freezing, QF) 및 완만동결(Slow Freezing, SF)을 하였으며 이에 따른 품질 변화를 실험하였다. 즉, 동결 방법에 따라서 40일간 저장하면서 저장기간 10일 간격으로 시료를 취하여 10oC에서 24시간 해동시킨 후 이화학 분석 및 미생물 실험을 실시하였다. 그 결과 모든 시료군에 대해서 환원당은 저장 10일째 급격히 증가한 후 저장기간에 따라 지속적으로 증가하였으나, 그 증가폭은 크지 않았다. 산도 및 색도 경우 저장기간에 따라 유의적 차이를 나타내지 않았다. 미생물의 경우 젖산균 및 효모 모두 저장기간이 증가할수록 감소하는 경향을 나타내었다. 젖산균의 경우 초기 젖산균이 4.1×107 CFU/mL에서 저장 20일째 QF는 3.6×106 CFU/mL, SF는1.8×104 CFU/mL로 나타나 QF 경우보다 SF경우 저장 중 젖산균에 대한 변화가 크게 나타났다. 효모의 경우에서도 동일한 변화를 나타내었다 이러한 결과 냉동 방법과 기간에 따라 막걸리 내 미생물의 생육을 제어하는 가능성을 알아볼 수 있었다.

This study was conducted to investigate the survival rate of frozen-thawed spermatozoa in equine by glycerol concentration and freezing speed. Two stallions (1 Thoroughbred-13 year old and 1 Arab-7 year old) bred in Korea Racing authority was examined for 1 times in a couple of weeks. Semen was collected by condom method standing heated mare and were centrifuged 650 g for 15 min. and isolated the seminal plasma. Thick fraction of semen was diluted EDTA-Lactose-egg yolk diluents to 1:1 and contained in 0.5 ml straw as 6～14×107cells/ml. Final concentrations of glycerol were 3, 5 and 7% in cryopreseved diluents and added 4 times for 2 hours equilibration. For the freezing, equilibrated straws were located 3 or 5 cm above LN2 gas for 5 or 10 min. Survival rates of pre-frozen sperm were 65.0±13.2%, 68.3±10.4%, 66.7±11.5% and post-frozen were 53.3±23.1%, 45.0±15.0%, 50.0±18.0% in 3, 5, 7% glycerol concentration, respectively. There was no difference between glycerol concentrations. Survival rates of frozen-thawed sperm on freezing speed were 36.7±10.4%, 40.0±7.1%, 30.0±13.2% at 3 cm－5 min and 33.3±11.5%, 31.7± 2.9%, 21.7±10.4% at 3 cm－10 min in 3, 5, 7% glycerol concentration, respectively. Survival rates of frozen-thawed sperm on freezing speed were 43.3±15.3%, 32.0±17.9%, 22.3±15.7% at 5cm－5 min and were 47.5±15.0%, 43.3±12.6%, 48.3±15.3% at 5cm－10 min in 3, 5, 7% glycerol concentration, respectively. There were significantly different between groups (p<0.05). These results suggest that glycerol concentration did not affect cryopreservation of stallion semen within 3～7% but freezing speed affects. In our experiment, the best cryopreservation condition was at 5 cm above LN2 gas for 10 min for pre-freezing and 7% of glycerol concentration. These results lead to commercial AI with frozen-thawed stallion semen.

This study was performed to investigate the characteristics within ages and freezing tolerance of spermatozoa in Jindo Dog. Experimental animals were selected 12 herds within 1～8 year’s old and collected semen for 2 times in a week. Collected semen was evaluated whole volume and sperm number with CASA system (SIAS, Medical Supply, Korea). Then seminal plasma were separated and diluted with modified Tris-egg yolk extender and added 4, 6 and 8% glycerol for 4 times to final concentration and equilibrated for 1.5 hrs. Before and after freezing, equilibrated semen were evaluated the survival rates. Total volume of sperm at 1～2 year old group is as 5.2×108 cells/ ml largest and there were no significance among groups. The motility of 1～2 year old group is highest as 90.9% and there were significance among groups. Abnormal sperm showed similar among groups. The survival rate in terms of pre-freezing and post-freezing were decreased all levels of glycerol and reveled 87.0% to 64.5% in 4%, 87.5% to 51.9% in 6% and 73.4% to 29.7% in 8%, there were significant difference among the groups (p<0.05). These results suggest that the optimal sperm-freezing methods in Jindo Dog are utilized with modified Tris egg-yolk extender with 4% glycerol and were improve the reproductive activity by these methods.

This studies were conducted to investigate the survival rate of frozen-thawed spermatozoa of Jindo Dog by monosaccharide and freezing rates. Experimental animals were prepared 12 males within 1~8 year's old and collected once in a couple of weeks by digital manuplation methods. Collected semen was diluted 1:1 with Tris-egg yolk extender and added 4, 6 or 8% of glycerol and none, 4 mM glucose or 4 mM fructose as cryoprotectant and was equilibrated for 2 hrs in . In monosaccharide groups, the freezing rate was 5 cm-5 min. above . The survival rates without monosaccharide were , , in 4, 6 or 8% glycerol, respectively. In addition of glucose, the survival rates were , , in 4, 6 or 8% glycerol, respectively and in fructose, were , , in 4, 6 or 8% glycerol, respectively. There showed significantly different between glycerol groups and monosaccharides groups (p<0.05). The survival rates of freezing rate in 5 cm-5 min. group was , , and in 10 cm-10 min. group was , , in 4, 6 or 8% glycerol, respectively. There were significantly different between freezing rates (p<0.05). These results suggest that the addition of fructose with 6%-glycerol and slow freezing improve the survival of frozen-thawed sperm in Jindo Dog.

도로는 다양한 재료와 단면으로 구성된 구조물이기 때문에 계절적 및 재료 물성특성 뿐만 아니라 포장체 각 층의 구조적 적정성 또는 지지력 정도를 파악하는 것이 무엇보다 중요하다. 현재, 기존 동상방지층 설계법에 따르면, 동상방지층은 포장체의 구조적 적정성과는 무관하게 온도조건에 따른 동결깊이에 따라 일률적으로 결정되고 있다. 이러한 동결깊이를 포장구조설계에 적용하다 보니 포장 설계의 부실 내지는 과다설계의 우려가 있다. 이에 본 논문은 2m 이하 저성토부, 절토부 및 절성경계부 등을 구분하여 포장 하부층에서 동상방지층의 구조적 역할을 규명하기 위하여 Falling Weight Deflectormeter(FWD) 시험을 수행하였다. 전국 10개 현장에 대하여 보조기층면에서 FWD 시험을 수행하였다. 각각의 단면은 동방방지층이 있는 구간과 없는 구간으로 구분하여 실험을 수행하였다. 이번 현장 실험을 통하여 동상방지층이 있는 경우가 없는 경우에 비해 처짐량이 작게 측정되어 동상방지층이 포장체에서 구조적 역할을 담당하는 것을 확인 할 수 있었다. 특히 절토부에서 동상방지층이 있는 경우에 처짐량 값이 약 15~55% 정도 감소하며, 절성경계부에서 동상방지층이 있는 경우에 처짐량 값이 약 11~64%, 저성토부의 경우 약 2~38%정도 감소하는 것으로 나타났다. SCI를 이용하여 분석한 결과, 동상방지층의 두께가 전체 포장체의 구조적인 능력에 약 24% 차지하는 것으로 나타났다. 피로수명은 동상방지층이 있는 구간이 없는 구간에 비해 약 2배 높은 결과를 보여 피로균열 저항성을 증진시키는 것으로 나타났다. 이것은 동상방지층이 포장체에서 구조적인 역할을 한다는 의미이다.

겨울에는 시베리아기단의 영향으로 한랭 건조한 대륙성 고기압의 영향을 받아 춥고 건조하여 1월 평균기온이 -6~-7℃의 영하의 온도로 낮아져 지반동결시 수분이동으로 동상현상이 발생하여 도로의 불균형 동결팽창을 초래하며, 결국 포장체를 파손시킨다. 동상발생시 토립자는 모관력에 의해 지하수를 흡수하여 아이스렌즈를 형성하며, 이 모관 흡수력은 토립자의 크기에 영향을 받는다. 도로는 다양한 재료와 단면으로 구성된 구조물이기 때문에 환경성과 재료 물성뿐만 아니라 포장체 각 층의 구조적 적정성 또는 지지력을 파악하는 것이 무엇보다 중요하다. 현재 기존 동상방지층 설계법에 따르면, 동상방지층은 포장체의 구조적 적정성과는 무관하게 온도 조건에 따른 동결깊이에 따라 일률적으로 결정되고 있다. 이러한 동결깊이를 포장구조설계에 적용함으로써 포장의 과다설계 우려가 있다. 따라서 본 논문에서는 도로 동상방지층의 효용성 검증 및 설치기준 확립을 위해 실내동결시스템을 활용하여 도로 노상토의 동상 특성에 대한 민감성을 판별, 도로건설 현장 노상토에 대한 역학적 특성과 실내 동결 시험을 수행하였으며, 외부 동결온도의 지속 조건에 대한 시료의 온도변화, 동상팽창압, 동상팽창량, 부동수분 등의 결과값을 통하여 동결 과정에 따른 지반공학적 특성을 평가하였다.

To enhance the embryo preservation technology and better application of embryo transfer technique to the field (dairy science or animal reproduction. etc.), we examined the viabilities of bovine embryos produced in vitro and in vivo after cryopreservation according to their developmental stage and thawing temperature. Bovine embryos from in vivo/vitro fertilization (Hanwoo) were examined at day 7, 8, and 9. Survival rates and total cell numbers of in vivo fertilized embryos were as follows: morulae 68.8% and ; blastocysts 80.5% and ; expanded blastocysts 77.4% and , respectively. Rates of embryo development for blastocysts and expanded blastocysts after thawing were significantly higher than that of morula stage embryos (p<0.05). While survival rates of in vitro fertilized embryos according to developmental stage showed no significant difference among groups (morula 67.9%; blastocyst 74.3%; and expanded blastocyst 79.4%), total cell numbers were significantly lower than those of other groups (morula ; blastocyst ; and expanded blastocyst ) For the viability according to thawing temperature, survival rate was higher in .

본 연구의 목적은 현재 국내에서 적용되고 있는 동결지수와 동결깊이 산정의 타당성을 검증하는 것이다. 이에 국내에서 운영되고 있는 LTPP-SPS(Long Term Pavement Performance- Specific Pavement Study)구간의 대기온도와 포장체 깊이별 온도 데이터를 이용하여 동결지수와 동결깊이를 산정하였으며, 기존 계산식을 이용하여 산출된 동결깊이와 비교 분석하였다. 또한 이 구간들의 인근 측후소 데이터를 이용하여 연도별 동결지수 추이를 분석하였다. 본 연구 결과, 지구온난화에 의해 1987년 이후에는 동결지수가 급격히 감소하였으며, 현재 도로설계 시 적용되고 있는 동결지수가 과하게 적용되고 있음을 알 수 있었다. 이에 동결지수 산정 시 현행 30년 데이터를 토대로 산정하기보다 기후가 변화된 시점부터 현재까지의 추이를 분석하는 것이 바람직 할 것으로 사료된다. 또한 LTPP-SPS 구간에서 측정된 포장체 깊이별 온도데이터를 분석한 결과, 동상방지층이 설계되어 있는 3개구간에서 대체적으로 보조기층까지만 동결이 발생하는 것으로 나타나, 동상방지층이 제 역할을 수행하지 못하는 것으로 나타났다. 실제 측정된 동결깊이와 계산식을 통해 산정된 동결깊이를 비교한 결과, 노상동결관입허용법을 이용한 설계방법이 실제 측정된 동결깊이와 가장 유사한 경향을 나타내었다.