• 한국동물번식학회:학술대회논문집
• /
• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
• /
• pp.81-81
• /
• 2003

돼지 정액의 동결보존은 돼지 정자가 내동성이 약한 특성을 가지고 있어 아직 만족할만한 결과를 얻지 못하여 액상정액을 이용하여 인공수정을 실시하고 있는 실정이다. 돼지 정액의 동결보존을 효과적으로 이룩한다면 돼지 인공수정의 효율성을 증대시킬 수 있고, 우수한 종돈의 보호 및 국제적 돼지 품종의 교류를 증진시킬 수 있을 것이다. 본 실험은 돼지 정액 동결시 동결온도와 시간, 항산화제로 알려진 Taurine의 농도별 첨가, 동해보호제 및 융해조건이 돼지 정액의 동결 융해후 생존성에 미치는 영향을 검토하고자 수행되었다.

• Journal of Animal Science and Technology
• /
• 제54권4호
• /
• pp.283-290
• /
• 2012

제주흑우의 동결 정액 제조시 동결 보존액에 첨가한 taurine과 hypotaurine은 동결 융해 후 정자의 운동성, 생존성 그리고 정자막 온전성을 개선시키는 것을 알 수 있었다. Taurine과 hypotaurine의 첨가는 유의적으로 높은 운동성, 생존성을 보였으며(p<0.05), 특히 hypotaurine은 다른 실험구에 비하여 유의적으로 높은 정자막 온전성을 나타냈다(p<0.05). 뿐만 아니라, hypotaurine은 유의적으로 높은 F pattern 비율을 유지하였으며(p<0.05), 이들 항산화물질을 첨가한 실험구에서는 대조구에 비하여 유의적으로 낮은 AR pattern을 나타내어(p<0.05) 동결로 기인된 수정능획득 유사 상태 정자의 비율을 유의적으로 감소시켜 조기 첨체반응 비율을 감소시켰다. 정자의 난자내 침투능력에 있어서 모든 처리구에서 대조구보다 높은 웅성전핵 형성율과 SFI를 나타냈으며, hypotaurine의 처리는 가장 높은 침투능력을 나타냈으나 처리구간의 유의적 차이는 나타나지 않았다. 본 연구결과는 멸실위험의 토종가축 생식세포 및 유전자원 보존과 토종가축 육성을 위한 번식 증대를 위한 제주흑우 동결정액 제조에 중요한 이용 방법이 될 것이며, 동결 융해 후 정자의 기능 개선을 위한 다양한 희석제 및 첨가제를 활용한 연구가 필요할 것으로 사료된다.

• 한국수정란이식학회지
• /
• 제24권3호
• /
• pp.199-205
• /
• 2009

This study was carried out to investigate the effective genetic resources preservation system using the frozen boar semen. The porcine oocytes were matured for 44 hours in NCSU-23 medium with or without 10% Porcine Follicle Fluid (PFF), 0.5 ${\mu}g/ml$ porcine FSH, 0.5 ${\mu}g/ml$ equine LH, 1.0 ${\mu}g/ml$ 17 $\beta$-estradiol ($E_2$) and 10 ng/ml Epidermal Growth Factor (EGF) under mineral oil at $38.5^{\circ}C$ in humidified atmosphere of 5% $CO_2$ in air. After 44 h of culture, the oocytes were inseminated with frozen-thawed semen and fresh semen prepared with mTBM medium for 6 h. Later, set of 50 presumptive zygotes were transferred into 4-well dish (500 ${\mu}l$) of IVC medium. for embryos freezing, slow-freezing and vitrification methods were used as a cryopreservation. Differences among treatments were analyzed using General Linear Model Procedure by SAS Package (version 6.12) differences were considered significant when p<0.05. Following IVF and IVC, the rates of cleavage and blastocysts formation were significantly higher (p<0.05) in hormone supplemented group than that of hormone-free group (25.7 vs, 12.1). The development rates to cleavage and blastocysts were significantly higher in PZM-5 group than NCSU-23 group (60.3%, 46.6% vs 27.4%, 11.1%). Further improvement was achieved when PZM-5 was supplemented with FBS. Cleavage rates was significantly higher in fresh semen source group than frozen semen (66.7% vs 43.7%). However in blastocysts rates was similar two groups. Post-thaw survival rates of embryos were 1.2% and 2.2% in slow-frezing and vitrification groups, respectively. The results of our study suggest that it is still possible to improve the culture conditions and boar semen cryopreservation for enhance reproductive technology and animal genetic resources conservation.

• 한국수정란이식학회지
• /
• 제32권3호
• /
• pp.235-241
• /
• 2017

가축유전자원으로서 흑염소 정액은 가축의 증식 및 유전적 개량을 위하여 동결정액으로서 보존될 필요성이 높으나 아직까지 연구 결과가 많이 누적되어 있지 않은 것으로 판단된다. 국내의 흑염소 동결유전자원의 생산성과 효율성 분석을 위하여 가축유전자원센터에서 보유 중인 흑염소 3 계통을 이용하여 전기자극방법에 의하여 채취된 정자와 정소상체유래 정자를 동결 보존하였다. 동결 전 Triladyl 난황희석액을 이용하여 흑염소의 정액과 혼합하여 $17^{\circ}C$에서 2시간 보존하면 응고현상을 관찰할 수 있었으며 42.9%의 비율로 난황이 변화한다는 것을 육안으로 관찰할 수 있었다. 정장 제거용 배양액으로 전기자극 유래 정자를 세정 후 동결 보존하면 융해 후 정자 생존율이 정액 세정용 배양액 $53.8{\pm}5.2%$로 나타났으며 정소상체 정자는 $74.6{\pm}10.6%$로 유의적으로 높게 관찰되었다. 융해된 정자의 장수성을 $17^{\circ}C$에서 조사해 보면 정소상체 유래 정자가 전기자극 유래 정자보다 우수한 것으로 조사되었다. 그러므로, 염소 동결 유전자원을 보존하는 방법으로 정소상체 정자는 활용성이 인정되며, 염소의 개량과 선발을 위하여 농가에서 활용 가능한 동결정액의 생산과 융해 후 생존성 증진을 위하여 더 많은 동결 보존 연구가 필요하다고 판단된다.

• 한국가축번식학회지
• /
• 제26권2호
• /
• pp.119-124
• /
• 2002

본 연구는 돼지에서 동결정액을 이용한 번식능력을 개선하기 위한 동결정액 포장재료의 효과를 구명하기 위하여 실시하였다. 본 시험에는 축산기술연구소 종축개량부 (충남, 성환)의 인공수정센터에서 사육중인 종모돈이 사용되었다. 기존의 돼지 동결정액 포장방법인 maxi-straw 동결정액 포장방법과 5$m\ell$빈 cryogenic-vial 및 aluminum-pack 포장방법을 비교한 결과 cryogenic-vial로 포장하여 액체질소 상단 15cm에서 동결한 후 52$^{\circ}C$ water bath에서 190초 융해한 방법이 기존의 maxi-straw 방법과 비슷한 결과를 나타내었다. Cyogenic-vial 포장방법의 동결-응해 방법을 설정하기 위하여 융해시간을 달리하여 시험한 결과 액체질소 상단 15cm에서 동결하고 52$^{\circ}C$에서 190초 간 융해하였을 때 정자운동성이 120초 및 150초 응해시 보다 우수하였다 (P<0.05). 그러나 정상첨체비율은 응해시간 간에 차이가 없었다. 52$^{\circ}C$에서 45초간 응해 한 maxi-straw 포장방법과 52$^{\circ}C$에서 190초 융해한 cryogenic-vial 포장방법간에 정액성상을 비교한 결과 총정자운동성과 정자의 빠르기는 maxi-straw가 우수하였다 (P<0.05). 그러나 직진성과 정상첨체비율은 두 포장방법간에 차이가 없었다. 동결정액 포장방법별 인공수정시 번식성적은 maxi-straw 동결정액이 cryogenic-vial 동결정 액보다 수태율, 분만율, 그리고 산자수가 높았으나 통계적 유의차는 인정되지 않았다. 이상의 결과를 종합해 보면 cryogenic-vial 포장 방법의 동결 및 음해방법을 좀 더 연구개발하면 기존의 maxi-straw포장방법을 대체하여 실용화 할 수 있을 것으로 사료된다.

• 한국수정란이식학회지
• /
• 제20권1호
• /
• pp.43-48
• /
• 2005

본 실험은 돼지정액의 동결보존을 위한 희석액과 냉각속도 및 동해방지제의 적정농도를 결정하기 위해 실시하였으며, 얻어진 결과는 다음과 같다. 1. $25^{\circ}C$에서 $5^{\circ}C$까지의 냉각속도에서는 LEY 희석액에서 분당 $0.17\~0.22^{\circ}C$로 냉각하는 것이 생존율과 정상 첨체율에서 가장 높은 결과를 얻었다. 2. LEY 희석액이 BF5와 M-Soejima 희석액보다 정자를 동해로부터 보호하는 능력이 우수하였다. 3. LEY 희석액에 첨가하는 glycerol의 농도는 3 또는 $4\%$가 $2\%$의 glycerol을 첨가한 것보다 4. LEY 희석액에 15mM의 fructose와 $3\%$ 또는 $4\%$의 glycerol 첨가구가 가장 높은 생존율과 정상첨체율을 나나냈다(p<0.001).

• Reproductive and Developmental Biology
• /
• 제29권3호
• /
• pp.149-154
• /
• 2005

The purpose of this study was the analysis of sperm ability in Specific Pathogen Free (SPE) miniature pig for production of bio-organ. The collected semen was diluted with extender and stored at $17^{\circ}C$t for up to 7 days. The semen samples were evaluated at 0, 1, 3, 5, and 7 days of storage for analysis of sperm ability. Sperm ability was evaluated by examining viability, progressive motility, sperm abnormality and intensity of the sperm membrane. Also, the semen was processed according to the convenient freezing method, and frozen-thawed sperm was evaluated by examining viability, capacitation and acrosome reaction using chlortetracycline (CTC) staining. Motility of spermatozoa of SPF miniature pig was significantly (P<0.05) lower on 3 days or later compared to the Duroc, Yorkshire and Landrace in domestic boar. The percentage of abnormal spermatozoa of Landrace were significantly (P<0.05) higher than in SPF miniature pig, Duroc and Yorkshire that had a similar percentage on 5 or 7 days of sperm storage. The percentage of spermatozoa with coiled tail decreased during the storage period but there were no significant difference. On the other hand, viability of frozen-thawed spermatozoa had a significantly (P<0.05) lower in SPF miniature pig than in other domestic boars. CTC patterns had no significant difference, but SPF miniature pig had higher percentage of capacitated spermatozoa and lower percentage of acrosome-reacted it than domestic boars. Therefore, this study suggest that it is necessary to develop the suitable extender and freezing methods methods for the high viable rate and fertilizing ability in vitro.

• 한국수정란이식학회지
• /
• 제28권2호
• /
• pp.87-93
• /
• 2013

The efficiency of artificial insemination (AI) for horses remains unsatisfactory. It is mainly because each process of AI causes a detrimental effect on semen quality. To sustain quality of semen properly, several factors including libido of stallions and sperm damage during sperm processing and preservation should be considered. Stallions with decent libido produce a high ratio of sperm to seminal plasma in their ejaculates, which is the ideal semen composition for maintaining sperm quality. Thus, to maximize the fertility rate upon AI, stallions should be appropriately managed to enhance their libido. Seminal plasma should have a positive effect on horse fertility in the case of natural breeding, whereas the effects of seminal plasma on both sperm viability and quality in the context of AI remain controversial. Centrifugation of semen is performed during semen processing to remove seminal plasma and to isolate fine quality sperm from semen. However, the centrifugation process can also result in sperm loss and damage. To solve this problem, several different centrifugation techniques such as Cushion Fluid along with dual and single Androcoll-E$^{TM}$ were developed to minimize loss of sperm and to damage at the bottom of the pellet. Most recently, a new technique without centrifugation was developed with the purpose of separating sperm from semen. AI techniques have been advanced to deliver sperm to optimal region of female reproductive tract at perfect timing. Recombinant equine luteinizing hormone (reLH) and low dose insemination techniques have been developed to maximize both fertility rate and the efficiency of AI. Horse breeders should consider that the entire AI procedure should be optimized for each stallion due to variation in individual horses for a uniformed AI protocol.

• Asian-Australasian Journal of Animal Sciences
• /
• 제15권1호
• /
• pp.16-18
• /
• 2002

During the process of freezing, spermatozoa suffer cold shock which increases their susceptibility to lipid peroxidation which plays an important role in ageing of spermatozoa, shortening their life span and affecting the preservation of semen. An experiment was therefore conducted to study the effect of addition of natural antioxidants into semen diluents on the preservability of buffalo semen. Split semen samples were extended in milk egg yolk diluents fortified with vitamin E (MYE), vitamin C (MYC) and control group (MYO); Tris-egg yolk diluents fortified with vitamin E (TYE), vitamin C (TYC) and control group (TYO) and evaluated for their preservabilities at 4-7$^{\circ}C$ and $37^{\circ}C$. Overall least squares mean of percent motility observed after 0, 24, 48, 72 and 96 h of preservation at 4-7$^{\circ}C$ were 66.70, 54.00, 36.80, 21.90 and 12.50, respectively while the estimates for semen extended in MYE, MYC, MYO, TYE, TYC and TYO were 44.80, 42.70, 38.70, 36.00, 35.20 and 33.00 percent, respectively. The results showed that motility was significantly (p<0.01) affected by extender (extender-antioxidant combination) and preservation interval. Overall least squares mean percent motility observed after 0, 4, 8, 12 and 24 h of preservation at $37^{\circ}C$ were 68.50, 58.90, 45.00, 38.10 and 18.10 percent, respectively, while the estimates for semen extended in MYE, MYC, MYO, TYE, TYC and TYO were 48.20, 49.30, 46.80, 45.30, 42.30 and 42.50 percent, respectively. Extender and storage interval were found to be significantly (p<0.01) affecting spermatozoa motility on room temperature preservation. The results indicated that the incorporation of antioxidants, especially vitamin E, had beneficial effect on preservability of buffalo semen.

• 한국가축번식학회지
• /
• 제10권1호
• /
• pp.19-26
• /
• 1986

This experiment was carried out to investigate the effect of bovine serum ablumin (BSA), sugars, glycerol equilibration time, straw size and thawing method on the survival index and the morphology of frozen boar spermatozoa. The results obtained were summarized as follow: 1. When the semen frozen in BF5 dilutor as pellet form was thawed in BTS at 37$^{\circ}$and 50$^{\circ}C$, BF5 dilutor with fructose showed higher sperm survival index than that with dextrose, however, when the semen was thawed on dry test tube at 37$^{\circ}C$, BF5 dilutor with sucrose showed higher sperm survival index than with other sugars. 2 When the semen forzen in BF5 dilutor with straw and thawed at 37$^{\circ}C$, BF5 dilutor with dextrose showed higher sperm survival index than those with other sugars, and there was no difference in sperm survival index between 0.5 and 1.0 ml straws. 3. The sperm survival index of frozen sperm was significantly (P<0.05) improved due to addition of BSA (0.05%) to BF5 dilutor. 4. When the extended semen with BF5 dilutor contatining 0.01 to 0.05% of BSA was frozen in the straw, the semen without glycerol equilibration showed significantly (P<0.05) higher sperm survival index than those with 2, 4 and 6 hrs glycerol equilibration time. 5. The sperm frozen in BF5 dilutor with dextrose or fructose, sucrose and raffinose showed 77 to 88% in normal acrosome rate and no difference among sugars. 6. The frozen semen showed lower normal acrosome rate than the first and second diluted semen, whereas the frozen semen showed higher swollen, damaged and missing acrosome rate than the first and second diluted semen. 7. Damaged and missing acrosome rate of sperm head due to freezing was somewhat inhibited by addition of BSA (0.01 to 0.05) to the BF5 dilutor.