• 한국가축번식학회지
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• 제26권3호
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• pp.253-261
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• 2002

본 연구는 진도개 동결정액 제조기술을 정립하기 위하여 진도개 정액성상과 동결 전후 정액의 활력과 생존율 및 CASAs를 이용한 운동성 등에 대하여 조사하였고, 백구와 황구간, 개체간의 정액성상과 내동성을 비교, 조사하였다. 이상의 연구결과는 다음과 같다. 1. 총 63회 정액채취 후 신선정액의 평균 정액량 3.8 $m\ell$, 농도 145.6$\times$$10^{6}$$m\ell$, 총정자수 396.2$\times$08/$m\ell$, 전진운동율 79.7% 및 생존율 89.5% 였다. 황구와 백구간에는 황구가, 개체간에는 황구 2호가 정자농도, 총정자수, 전진운동정자율 및 생존율 등의 정액성상에서 유의적으로 우수하였다(P<0.05). 2. 동결전.후 정자의 전진운동율과 생존율을 46회 조사한 결과 동결전 73.5%와 82.3%를, 동결후 51.1%와 64.9%를 나타내 동결과정이 전진운동율과 생존율에 영향이 있었으며, 역시 황구와 백구간에는 황구가, 개체간에는 황구 2호가 동결전후 전진운동율과 생존율에서 유의적인 차이가 인정되었다 (P<0.05). 3. 동결.융해정자의 보다 객관적인 평가를 위하여 CASA system을 이용한 총 44회 평가한 결과, 생존율 65.6%, 전진운동율 54.8%, VAP 75.3 $\mu\textrm{m}$/sec, VCL 90.0 $\mu\textrm{m}$/sec, VSL 69.4 $\mu\textrm{m}$/sec 및 ALH 4.4 $\mu\textrm{m}$로 동결 융해 정액의 운동성은 양호하였고, 황구와 백구간의 운동성에는 유의적인 차이가 없었으나, 개체간에는 역시 황구 2호가 운동성이 우수하여 유의적인 차이가 인정되었다 (P<0.05). 4. 채취된 정액중 백구는 46%(13/28), 황구는 94%(33/35)가 동결정액 제조가 가능해 황구가 내동성이 좋았으며, 전체적으로 73%(46/63) 동결정액 제조가 가능하였다. 결론적으로 진도개의 동결정액을 제조하기 위하여 정액성상 및 동결 전후 운동성을 조사한 결과 동결정액 제조와 생산체계의 구축이 가능하였으며, 황구와 백구간, 개체간의 정액성상과 동결전후의 운동성에 차이가 인정되므로 정액성상과 내동성을 고려한 종견선발 체계가 필요하다고 사려되었다.

• 대한수의학회지
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• 제47권1호
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• pp.135-138
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• 2007

A 3 year-old female Afghan hound came to the Veterinary Referral Hospital of the College of Veterinary Medicine, Seoul National University for artificial insemination (AI) with frozen semen. In order to inseminate, semen was frozen in USA 3 years ago. Frozen semen was sent by air from Santiago to Seoul for AI. The stud died 2 years ago, so we could only use a limited amount of frozen semen in that estrus cycle. The number of total motile spermatozoa was $59.4{\times}10^6$and the total volume was 1.2 ml. The frozen spermatozoa were thawed in $70^{\circ}C$ water for 8 sec, which were then deposited at the bilateral uterine horns by a surgical method. The number of corpus luteum was 6. Sixty days after artificial insemination resulted in the birth of 4 puppies, all of which are alive and healthy.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.92-92
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• 2002

Considerable attention has been focused on the cryopreservation of semen and estrus induction in dog, as consequence of poor productivity caused by long anestrus period, in order to enhance the productivity of youngs and to preserve the breeds. The objectives of this study were to evaluate semen quality after cryopreservation and to evaluate the Pregnancy rate after insemination (AI). Fifty infertilie dogs (age 2∼3 years) were selected for the study and divided into three different estrus induction treatment groups. Group 1: dogs (n=15) were given clomifene (0.1 mg/kg) orally for five days at 12 hr intervals. Group 2: dogs (n=15) were given bromocriptine (50 $\mu\textrm{g}$/kg) orally for five days at 12 hr intervals, followed by single injection intravenously of 500 IU GnRH (Group 3, n=20) when pro-estrus occurred. The rates of pregnancy in estrus inducted dogs mated naturally compared to those inseminated artificially with ejaculated fresh semen and frozen-thawed semen. Estrus detection was performed using the method of vaginal smear and confirmed by the plasma progesterone assay. The ejaculated semen to freeze was exposed to a mixture of Tris extender with cryoprotectant (Trisma, 81 mM: TES, 209 mM: citric acid, 6 mM; glucose, 5 mM; glycerol, 8%) and cryopreserved gradually by slow-cooling at 17 cm above the surface of liquid nitrogen (LN$_2$) for 23 min. The motility of frozen-thawed spermatozoa was assessed by phase-contrast microscopy. To assess their viability and acrosome content, spermatozoa were stained with a vital stain and Fluorescence conjugated lectin Pisum Savitum Agglutinin (FITC/PAS), respectively. Pregnancy was confirmed by ultrasonograpy on day 25, 35 and 55 post insemination. The use of fresh semen, the pregnancy rates were observed 66.6, 66.6, 75.0 and 83.3% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. The use of frozen-thawed semen, the pregnancy rates were observed 66.6, 33.3, 50.0 and 60.0% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. No difference was observed in the number of offspring produced among natural estrus and treated groups inseminated with fresh or frozen-thawed semen. In conclusion, the pregnancy rate of dogs treated with a combination of GnRH and bromocriptine was more effective than use of clomifene or bromocriptine only. In addition, frozen-thawed semen can be used successfully far artificial insemination in dog.

• 한국임상수의학회지
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• 제18권1호
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• pp.44-47
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• 2001

Semen were collected from 9 male dogs and frozen by liquid nitrogengas. Frozen semen were thawed at 7$0^{\circ}C$ for 8 seconds. About $2{\timss}10^8$ sperm per insemination were inseminated to 10 bitches (3 Retrievers, 4 Chihuahuas, 1 Yorkshire Terriers, 1 Maltese, and 1 Poodle) at three and six days after the estimated peak of luteinizing hormone. For small breed dogs, uretero-renoscope (Kahl Storz, Germany, 12.5 Fr) was used for trans-cervical insemination, whereas cystoscope(Kahl Storz, Germany, 22Fr) was used for large breeds (Retrievers). Pregnancy was diagnosed by ultrasonography at 30 days after insemination. All of 3 Retrievers (100.0%) and 3 bitches of 7 small breed dogs (42.9%) were conceived (60.0% in total). This result indicated that trans-cervical insemination using endoscope is an effective method for AI with frozen semen not only for large breed dogs such as Retriever but also for small breeds.

• Asian-Australasian Journal of Animal Sciences
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• 제16권2호
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• pp.180-184
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• 2003

The intrauterine inseminator (IUI) was developed to provide the method of depositing dog semen into the uterine body instead of the vagina. The IUI consists of a vaginal endoscope, a balloon sheath, and injection catheter. When the endoscope is inserted into the vagina and the balloon expanded with air, the cervical os becomes visible so a injection catheter can be inserted through the cervix for deposition of the frozen-thawed semen. The efficacy of the IUI device was compared to intra-vaginal artificial insemination using semen that had been collected and frozen from pooled sperm-rich fraction of ejaculates collected from two Jindo dog donors. Aliquots of semen were extended with a Tris-egg yolk diluent, centrifuged, the seminal plasma removed, the pellet resuspended with the same diluent, and cooled to $5^{\circ}C$ over a 2 h period. A Tris-egg yolk-glycerol extender was added at $5^{\circ}C$; after 1 h, semen was loaded into 0.5 ml straws, and straws were frozen in LN vapor for 5 min, and immersed in LN for storage. The final sperm concentration for freezing was approximately $100{\times}10^{6}cells/ml$. The straws were thawed at $70^{\circ}C$ for precisely 6 sec, 1.5 ml Tris-egg yolk buffer at $38^{\circ}C$ added, and the 2 ml of thawed semen was used for a single insemination using the IUI device. Each bitch was inseminated at optimal insemination point, which was estimated by vaginal epithelial cells staining and progesterone concentration analysis. Use of the IUI device resulted in 21 of 26 females giving birth to 89 pups ($4.2{\pm}1.6$ pups per litter), while intra-vaginal AI resulted in 6 of 15 females whelping a total of 17 pups ($2.8{\pm}1.2$ pups per litter). We believe the IUI device is easier to use than previously described devices used for intrauterine insemination. In our experience the expansion of the balloon has a calming effect on the bitch that aids the inseminator. These results indicate that the IUI device was able to provide high fertility with 50 million frozen sperm per insemination and two inseminations.

• 한국수정란이식학회지
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• 제23권2호
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• pp.115-118
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• 2008

Glycerol is the cryoprotectant most frequently used to freeze semen in several of species. The objective of the present study was to compare the effect of three different glycerol concentrations (4, 6 or 8%, v/v) on frozen-thawed dog sperm survival rate. Ejaculates from 9 dogs collected by digital manipulation were pooled and assessed by macroscopic and microscopic criteria. Semen was divided into 3 aliquots, which were centrifuged and the sperm pellets rediluted with first Tris-glucose-citric acid extender. After 1 h cooling at $4^{\circ}C$, second extender containing 4, 6 or 8% glycerol was added, respectively. The semen was loaded into 0.25 ml straws and frozen and stored in liquid nitrogen and thawed. Sperm vigor, live:dead spermatozoa ratio using HOS test, and sperm morphology using $Spermac^{(R)}$ stain were evaluated. After thawing, there were no significant differences among groups in vigor, viability and morphology. In conclusion, the three glycerol concentrations (4, 6 or 8%) can be used successfully in cryopreservation of canine semen. Therefore the use of 4% glycerol in the extender has less toxic effect and reduces of freezing injuries.

• 한국수정란이식학회지
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• 제23권4호
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• pp.239-243
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• 2008

This study was conducted to investigate the effects of type of the sugar supplemented to the extender on the vigor, viability and intact acrosomal rates of frozen-thawed dog spermatozoa. The ejaculated semen was diluted with TRIS-citric acid extender containing 200mM TRIS, 73mM citric acid, 6% (v/v) glycerol, 20% (v/v) egg yolk, 1% (v/v) antibiotics (streptomycin/penicillin), 44 mM sugar, which was either glucose, fructose or glucose-fructose combination, and distilled water to make the final volume of 100ml. Extended semen samples were cooled at $4^{\circ}C$ for an hour, packaged in 0.25ml straws, equilibrated for 10 minutes in liquid nitrogen vapor, and frozen in liquid nitrogen. Samples were thawed by placing straws into $37^{\circ}C$ water for 120 seconds. After thawing, vigor, viability and intact acrosomal rates of frozen-thawed semen were compared according to type of sugar. No significant differences were observed between glucose and fructose groups. In addition, combination of the 2 sugars also did not show any significant differences in the vigor, viability and intact acrosomal rates. In conclusion, glucose and fructose were equally efficient as sugar supplements for freezing extender.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2001년도 추계학술대회 및 정기총회
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• pp.30-30
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• 2001

• 한국임상수의학회:학술대회논문집
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• 한국임상수의학회 2009년도 추계학술대회
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• pp.260-260
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• 2009

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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• pp.57-57
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• 2001

This study was carried out to investigate the general characteristics such as volume, sperm concentration, sperm motility, sperm abnormality on whole semen, RSP-S and RSP-T semen and fractional semen of small size dogs, and the effect of temperature and preservation time and cryoproservation on motility of whole and RSP-S and RSP- T semen. Multiple ejaculates were collected from small dogs by the digital manipulation of penis. 1. The volume per ejaculate semen, sperm of concentration and motility and abnormal sperm rate of 1st fractional semen were 0.65±0.09㎖, 4.52±0.35×10/sup 6/ cells/㎖, 15.64±3.85% and 5.50±0.62%. Also, 2nd fractional semen were 1.25±0.20㎖, 3.35±0.48×10/sup 6/cells/㎖, 96.25±4.65% and 4.24±0.46%. And 3rd fractional semen were 1.45±0.21㎖, 3.85±0.52×10/sup 6/cell/㎖, 92.82±4.24% and 4.66±0.58%, respectively. 2. The sperm of concentration and motility and abnormal sperm rates of whole, RSP-S and RSP-T semen were 5.45±0.82×10/sup 6/ cells/㎖, 95.55±4.65%, 4.58±0.45% and 4.82±0.36×10/sup 6/cells/㎖, 90.10±3.42%, 6.48±0.68% and 4.55±0.45× 10/sup 6/cells/㎖, 93.25±3.85%, 4.82±0.58%, respectively. 3. The motility of whole, RSP-S and RSP-T semen were higher at 4℃ than at 38℃. When preservation temperature was at 4℃, survival rates of RSP-S and RSP-T sperm were 97.54%-6.25% at 1-72 hrs, 97.40%-5.62% at 1-100 hrs, respectively. 4. The survival rates of slow and rapid frozen 2nd fraction, RSP-S and RSP-T semen were 67.3±4.45%, 88.8±4.46% and 46.4±3.84%, 74.4±4.20%, respectively. Survival rates was significantly higher in frozen RSP-S and RSP-T semen than that in control group(8.5±2.12%).