• 한국임상수의학회지
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• 제17권2호
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• pp.420-430
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• 2000

These studies were preformed to investigate the freezing conditions to achieve good post-thaw viability of spend and the practical methods of artificial insemination frozen canine semen. Semen were collected from nine male dogs which had been proved to be fertile in the past and the semen were treated for freezing procedure. Post-thaw motility and viability of canine sperm were evaluated to investigate individual tolerance of freezing, difference among freezing extenders, dif-ference among freezing equipments and freezing conditions, difference between fast and slow cooling rate, difference according to different glycerol concentration, effect of seeding on post-thaw viability, difference according to cutting part of straw, difference according to thawing temperatures, and dif-ference according to media added to thawed semen. Thawed semen for insemination were added with equal volnme of canine capacitation medium (CCM) and the volume of semen and the number per insemination were adjusted as 2-3 ml and $20-30 {\times}10^7,$ respectively. The semen were inseminated in vagina using balloon catheter and en17ryos were cellected from 9 to 11 days after the second Al to d determine fertilization.

• 한국수정란이식학회지
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• 제20권2호
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• pp.157-162
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• 2005

To find out the possible inefficiencies of artificial inseminators at rectovaginal insemination in cows, inseminators' skill were evaluated by controlling the semen thawing procedure adopted and by using the technique of dye deposition in the genital tract of slaughtered cows. This was followed by refreshment training for the inseminators. Thirty seven artificial insemination technicians regularly working in the government, cooperative and NGO (Non Government Organization) artificial insemination programmes at different places of Bangladesh were included in the study. Individual technicians were asked to thaw a semen straw and deposit dye in the genital tract of slaughtered cows following the procedures they would have adopted in their actual practices of insemination. The time and water temperature adopted by technicians were recorded and genital tract after sham artificial insemination was dissected to determine the site of dye deposition. Then, the inseminators took part in a three days intensive training program. The training program was ended up with the same tests for thawing frozen semen straw and dye deposition in the genital tract of slaughtered cows. At pre training evaluation, only $25\%\;and\;72\%\;(n=36)$ inseminators adopted co..ect thawing time and temperature, respectively. At post training evaluation, all inseminators thawed semen straws for proper time and temperature. At pretraining evaluation, $21(57\%),\;11 (30\%)\;and\;3(8\%)$ inseminators deposited dye at the body of uterus, in the vagina or in cervix, and into the horn of uterus, respectively. In $2(5\%)$ cases dye did not pass into the genital tract, instead back flowed through the space between the barrel of insemination gun and sheath. At post training evaluation, all inseminators successfully deposited dye in the body of uterus. Frequent evaluation of inseminators' skill and subsequent training would help improvement of the artificial insemination technicians' skill.

• 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.

• 한국임상수의학회지
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• 제16권2호
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• pp.375-380
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• 1999

This study was conducted to develop an intrauterine inseminator (IUI) to deposit of frozen semen into uterus and to evaluate the results obtained after artificial insemination by IUI. Two Japanese spitzs (2 to 4 years of age) were used as semen donors. Semen was collected by manual masturbation into sterile glass collection tubes and separated into 3 fractions with only the sperm-rich fractions retained for further examination. Sperm motility >70%, sperm concentration of 200 to $400{\times}10^6 cells/ml$$\times$g for 5 min and poured out the suspended solution, and then diluted with 2 ml Tris-buffer which was consisted of 2.4 g Tris, 1.4 g citric acid, 0.8 g glucose, 0.1 $\mu\textrm{g}$/ml streptomycin, 100 IU/ml penicillin, 20 ml egg yolk to 100 ml mili-Q water (Ext I) or supplemented with 8 ml glycerol and 1 ml Equex STM paste to 100 rnl (Ext II). The diluted semen was cooled to 5$^{\circ}C$ in cold room, where the temperature in the sample reached 5$^{\circ}C$. Two h after beginning the cooling procedure, 2 ml of Ext II, also at 5$^{\circ}C$, was added and mixed by gently reversing the tubes several times during 1 h. The final sperm concentration for freezing was approximately $50{\times}10^6 cells/ml$. After equilibration, the semen was loaded into 0.5 ml straw and frozen on the liquid nitrogen vapour in styrofoam box. The straws were thawed at 7$0^{\circ}C$ for precisely 6 sec. After thawing of each straw, the frozen semen can survived over 50% motility. All the females were inseminated twice with 1 ml of $25{\times}10^6 cells/ml$

• 농업과학연구
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• 제34권1호
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• pp.61-65
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• 2007

본 연구에서는 유산양에서 비번식계절과 번식계절에 각각 발정동기화를 유도하여 인공수정을 1차와 2차에 걸쳐 실시한 후 분만율을 조사하였다. 자아넨 유산양 암컷을 대상으로 5월과 6월 사이에 CIDR-S를 이용하여 발정을 유도하였고, 액상정액과 동결정액을 이용하여 인공수정을 실시하였다. CIDR-S를 이용한 발정유도시험에서는 유산양의 발정동기화가 90%이상을 기록하여 아주 양호한 것으로 나타났다. 그러나 분만 성적은 1차 인공수정에 의해서는 25%, 2차 인공수정에 의해서는 20%로 낮하게 나타났으며, 자연종부에 의해서는 100%의 분만율을 기록하였다. 본 연구의 결과 비번식계절에서 번식주기를 인위적으로 조절할 수 있는 가능성과 정액을 주입하는 적정한 시기를 예측할 수 있는 점을 시사해 주었으나 인공수정 방법과 정액의 저장방법에 대한 연구가 필요한 것으로 나타났다.

• 한국수정란이식학회지
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• 제25권2호
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• pp.89-92
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• 2010

We report herein the successful results of estrus induction, sperm cryopreservation and kids born by transcervical insemination of frozen-thawed semen in a Saanen goat. Flugestone acetate (FGA: 60 mg) was inserted into vagina for 15 days. The goat was intramuscularly injected with 400 IU PMSG and 200 IU hCG ($PG600^{(R)}$: Intervet, Korea) a day before withdrawal of the FGA sponge. Follicles and corpora lutea were identified on both ovaries by laparoscopy. Artificial insemination was performed 46 hours after removal of FGA sponge. The concentration of frozen-thawed semen was $3.975{\times}10^8/ml$ and 0.5 ml of frozen-thawed semen was transcervically inseminated into uterine body under anesthesia. Three kids, all females, were born 144 days after artificial insemination. This is the first report producing kids by transcervical insemination of frozen-thawed semen in a Saanen goat of which the estrus was induced by FGA sponges, PMSG and hCG during non-breeding season in Korea.

• Asian-Australasian Journal of Animal Sciences
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• 제15권5호
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• pp.633-636
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• 2002

Six adult Garole rams maintained under a semi-intensive system were used as semen donors for this study. Semen was collectied daily during the monsoon season with the help of an artificial vagina and examined for its quality characteristics. Ejaculates of thick consistency, rapid wave motion, ${\geq}80%$ motility and intense movement of motile spermatozoa were diluted at the rate of 1:1 with egg yolk McIllvaine glucose diluent at $30^{\circ}C$ in water bath. Estrus in ewes was detected by parading aproned rams of proven vigour at 12 h intervals. The ewes (54 Malpura and 23 Avikalin) in estrus were artificially inseminated with fresh diluted ram semen. The overall conception rate was 94.8%, (range 91.7 to 100%). The overall lambing percent was 80.5 with a range of 75.0 to 84.6%. There was no significant (p>0.05) difference in lambing and conception rate because of individual rams. Fertility was significantly lower (p<0.05) in ewes of less than two years and more than six years of age. Breed (Malpura and Avikalin) effect was not observed in conception and lambing rate (p>0.05). No significant difference (p>0.05) in birth weight and 12 month weight was observed between Garole${\times}$Avikalin and Garole${\times}$Malpura crossbred lambs but there was significant (p<0.05) difference at three month and six months body weight of both the crossbred lambs.

• 한국수정란이식학회지
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• 제24권4호
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• pp.289-292
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• 2009

Mongolia has 80% livestock of total agriculture industry, 170,000 farms are engaged, 2,500,000 of cows that were beef and dairy cows are raised. Despite of Mongolian has great application with milk, there are not clear differences between cow and dairy cattle, and the production of milk is also low. But the milk suppliers are varied (horse, sheep, goat, etc), so that the total milk production is 500 thousand ton per year. It's really considerable to improve the breed of owing to many problems with big differences among milk qualities. For carrying out for first year project, artificial insemination project was operated with 3rd grade Holstein semen that were imported from S. Korea, and initiation and field training were also carried out through appropriate AI technique we developed for Mongolia environment. Local information research and MOU conclusion were done with professor D. Altangerel in May $10^{th}{\sim}13^{th}$, 2009, and development for AI technique and AI equipments were supplied for Mongolia breeding and natural environment in July $10^{th}{\sim}17^{th}$ in 2009. All cows were treated by synchronization for AI. To do this, $PGF_{2\alpha}$ injection were treated for luteal phase cow, if it wouldn't work, try again after 11 days. After confirmation of estrus, AI and AI training were carried out with sperm injection in the uterus or cervix by rectum-vagina method which is common worldwide, the most effective artificial insemination technique. If cows were return to next estrus cycle, second AI was carried out about approximately 21 days after artificial insemination. After 2 months, all cows not showing return estrus should be taken pregnancy test. Every pregnant cow will be cared thoroughly. Total 48 cows administrated by $PGF_{2\alpha}$ for synchronization and after 48 hours 45 cows (93.8%) showing estrus were detected and then artificial inseminate them within who 8 cows (27.8%) showed return estrus. Therefore, Using $PGF_2{\alpha}$ for synchronization is effective to use for Mongolia breeding conditions. There are possibility of base for food production after all, including increase of livestock production in Mongolia by improvement of breeding cow with AI and embryo transfer project.

• Asian-Australasian Journal of Animal Sciences
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• 제5권3호
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• pp.439-447
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• 1992

Eight healthy Holstein bulls, 4-6 years old were used to study the effect of season of the year on the biophysical characteristics of semen. Semen was collected twice a week by AV (artificial vagina) over one-year period. The analyses revealed that all the basic seminal traits studied were differed significantly due to season, except the ejaculate volume and consistency and the percentage of swollen spermatozoa in a hypo-osmotic fructose-citrate solution. Ejaculates collected during hot summer season had significantly lower sperm motility, concentration and total counts, and higher percentage of dead spermatozoa than those collected during winter time. Warm spring had moderate semen quality. The temperature-humidity index was calculated and it was associated (p < 0.01) negatively with the ejaculate pH, sperm concentration and total counts, and positively with the % of dead sperms. Ejaculate volume, percentage of swollen spermatozoa, individual motilities did not correlate significantly with the change in temperature-humidity index values. The total live, motile spermatozoa per ejaculate during both the winter and spring seasons showed significant increase of about 37% and 32% respectively over the summer season. Also, rectal temperatures of the bulls were elevated during the hot summer season, while the values of blood hemoglobin and packed-cell volume were decreased.

• 한국가축번식학회지
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• 제4권1호
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• pp.75-82
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• 1980

This experiment was carried out to improve a simple and effective procedure of egg transfer which is considered to be the most useful technique for the improvementand proliferation of domestic animals. Several experiment procedures such as superovulation, surgical recovery of ovulated egg, in vitro capacitation of ejaculated spermatozoa, in vitro fertilization and culture of embryo were conducted. The results obtained in this experiment were summarized as follows: 1. In rabbits treated with PMS in combination with estradiol and HCG, 10 to 43 eggs were obtained in one rabbit and the average ovulation number of 20 rabbits was 21 eggs. 2. Six to 24 eggs (average 13 eggs) were recovered from the removed oviducts by the methods of flushing technique and the average recovery rate of 20 rabbits was 61.9 percent. 3. Most rabbit spermatozoa ejaculated by artificial vagina were capacitated in vitro by the culture of the spermatozoa with PBI medium at 37$^{\circ}C$ for 4 hours after removal of seminal plasma. 4. Normal fetilization following in vitro fetilization was observed in 88 (42.7%) of 206 eggs. 5. The number of eggs developed to 2-, 4-, and 8-cell stage following in vitro culture with PBI medium at 37$^{\circ}C$ was 26 (70.3%), 20 (54.1%) and 17 (45.9%) of 37 eggs used for in vitro culture.