• Journal of Embryo Transfer
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• v.26 no.1
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• pp.13-17
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• 2011

Research in the area of equine artificial insemination (AI) has led to its increased application in field trials. However, procedures for equine semen collection, cooling and freezing of semen and artificial insemination need further improvement. In experiment 1, we investigated the percentage of total motility (TM) and progressive motility (PM) of sperms at after-collection, cooled-diluted, cooled-transported or frozen-thawed semen. In experiment 2, mares were inseminated with either cooled-diluted, cooled-transported or frozen-thawed semen. In experiment 3, we examined the effect of buffer (skim-milk extender), which was infused into the uterus at the time of AI with frozen-thawed semen. In experiment 4, we compared AI pregnancy rates for mares ovulating spontaneously versus after treatment with hCG. In experiment 1, the average percentage of TM was decreased from 75.3% to 14.4% at the stage of after-collection to frozen-thawed semen (p<0.05). The average percentage of PM was 58.2% and 59.6% at after-collection and cooled-diluted, but it was significantly increased 71.7% after frozen-thawed (p<0.05). In experiment 2, the pregnancy rates after AI using cooled-diluted, cooled-transported and frozen-thawed semen were 60%, 50% and 37.5%, respectively, and similar among treatments. In experiment 3, the pregnancy rate of mares infused with buffer at AI was 40% which was higher than that with no buffer (10%). In experiment 4, the pregnancy rates of mares were similar between ovulated spontaneously (25%) and ovulated with hCG (50%). The results suggest that equine semen that has been cooled-diluted, cooled-transported or frozen can be successfully used to establish AI, pregnancy and foal production. Also, the pregnancy rates after AI can be increased by infusing buffer into the uterus at AI or by inducing ovulation with hCG, but further study is need.

• Journal of Embryo Transfer
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• v.25 no.2
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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.

• Journal of Animal Science and Technology
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• v.56 no.3
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• pp.8.1-8.6
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• 2014

Background: Artificial insemination (AI) can serve as a powerful tool to the sheep owners for making rapid genetic progress of their flock. The AI in sheep is mostly performed using fresh semen with two reasons i) lambing rate following trans-cervical AI with frozen semen is limited by the inability of frozen-thawed sperm to transit the cervix and ii) the need of circumventing the cervical barrier through laparoscope aided intrauterine AI. Therefore, AI with frozen-thawed semen is not as widespread in sheep as it is in other domestic species. However, to get maximum benefits through the use of AI, frozen-thawed semen is a prerequisite because instead of high fertility, the short shelf life of fresh semen coupled with a limitation on the number of insemination doses achievable per unit time restricts the widespread use of individual sires. Therefore, in order to enhance lambing rate, a total of 240 trans-cervical artificial inseminations with frozen-thawed semen were performed in Bharat Merino ewes during autumn season either once in the evening (G-I, 10 h after onset of estrus, n = 100) or twice (G-II, 14 h and 22 h after onset of estrus, n = 140) i.e. once in the morning and again in the evening. Results: The pregnancy rate (proportion of pregnant ewes confirmed by ultrasonography at day 40) and lambing rate (proportion of ewes lambed) were higher in G-II as compared to G-I (26.4 vs 20% and 19.3 vs 10%, respectively). The difference in lambing rates was statistically (P < 0.05) significant. The depth of insemination within cervico-uterine tract had no significant effect on pregnancy and lambing rates. Conclusions: The results indicate that lambing rate in sheep following TCAI with frozen-thawed semen was significantly influenced by time of inseminations. Two inseminations after 14 and 22 h of onset of estrus enhanced the lambing rates of Bharat Merino sheep as compare to single insemination after 10 h of onset of estrus. The TCAI technique with frozen-thawed ram semen is promising and may serve as a valuable tool for genetic improvement of sheep breeds. Research efforts are going on worldwide to overcome the poor fertility following TCAI with frozen-thawed semen.

• Korean Journal of Animal Reproduction
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• v.26 no.4
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• pp.339-345
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• 2002

The present study was carried out to investigate the effects of season on semen characteristics, frozen-thawed sperm viability and testosterone concentration in Yorkshire boars. There were no significant differences in the semen volume and sperm concentration on Yorkshire boars among spring, summer, autumn and winter. However, the pH of sperm-rich and sperm-poor fractions in winter season was higher than in spring, summer and autumn season in Yorkshlre boars. Sperm motiliy and normal acrosome of raw semen in Yorkshire boars did not differ significantly among spring, summer, autumn and winter. However, motility and normal acrosome of frozen-thawed sperm were higher in spring season than in summer, autumn and winter. Serum testosterone concentrations in Yorkshire were higher in spring than summer, autumn and winter. In conclusion, we found out that serum testosterone concentrations were very important role for frozen-thawed sperm viability in Yorkshire boars.

• Korean Journal of Animal Reproduction
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• v.10 no.1
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• pp.19-26
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• 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.

• Animal Bioscience
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• v.36 no.12
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• pp.1821-1830
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• 2023

Objective: This study investigated the effect of adding seminal plasma to frozen-thawed semen on the quality of sperm and pregnancy following insemination in dromedary camels. Methods: In experiment 1, the frozen-thawed semen from 9 collections (3 bulls) was further diluted with either the base extender or homologous seminal plasma (HSP). In the second experiment, a pooled sample of frozen-thawed semen was diluted with either seminal plasma from another three bulls. Live percentage, total and progressive motility, functional and acrosome integrity, and sperm kinematics were evaluated at 15, 60, and 120 minutes post-thawing and compared to the non-treated control. In experiment 3, frozen semen was used to inseminate camels in the following experimental groups: 1-Single insemination with double dose undiluted frozen semen (n = 9); 2-Re-insemination in 6 hours with undiluted semen (n = 13); 3-Single insemination with HSP treated sperm (n = 14). Results: Frozen-thawed sperm diluted in HSP or the non-homologous seminal plasma from Bull C indicated an improvement in all parameters after 1 hour post-thawing incubation (p<0.05). The proportion of total and progressively motile sperm did not drop significantly at 60 minutes post-thawing when diluted with the seminal plasma of Bull C (p>0.05). Double insemination with nontreated sperm and single insemination with HSP-treated sperm resulted in similar pregnancy rates (15.3% vs 21.4%, p>0.05). None of the camels conceived with double-dose single insemination of nontreated sperm. Conclusion: Seminal plasma improves sperm longevity and motility after thawing in dromedary camel with a significant between-bull variation in effect. Low post-thaw sperm longevity might be the cause behind the low pregnancy rates in frozen semen insemination of dromedary camels.

• Journal of Embryo Transfer
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• v.23 no.3
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• pp.161-166
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• 2008

The techniques for the collection, cooling and freezing of semen and artificial insemination of horses are not fully understood in Korea. We investigated the percentages of total motile (TM) and progressively motile (PM) sperms after the collection, cooling and freezing of stallion semen. The average volume of semen was 167 ml in Thoroughbred and 68 ml in Arab. The average numbers of spermatozoa in Thoroughbred and Arab were $104\times10^6/ml$ and $86\times10^6/ml$ respectively. The average percentages of TM and PM were 82.3% and 88.6% in Thoroughbred, and 61.4% and 82.6% in Arab, respectively. The average percentage of TM at 4 hr after cooling at $5^{\circ}C$ was significantly lower than that at 0 hr ($30.0\pm4.1%\;vs.\;78.0\pm2.5%,\;p<0.05$), but the percentage of PM was similar between 66.5 and 73.2% at 0, 1, and 4hr. The average percentage of frozen-thawed Thoroughbred semen frozen in MFR5 extender was 56.2%, which was significantly higher than that of the semen frozen in LE extender (average 32.9%, p<0.05). The percentage of TM in Arab was similar for semen frozen in MFR5 extender and LE extender (18.2% and 21.2%, respectively), but the percentage of PM was significantly higher in sperm frozen in MFR5 extender than in sperm frozen in LE extender (69.0% vs. 36.4%, p<0.05). Four mares were artificially inseminated by Thoroughbred frozen-thawed semen and one of them fertilized at 11 day after artificial insemination. In this study, the collection, cooling and freezing of equine semen were possible under domestic conditions.

• Journal of Embryo Transfer
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• v.14 no.2
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• pp.131-137
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• 1999

This experiment was carried out to investigate the effects of osmolarity of thawing diluents, seminal plasma added in thawing diluents on the sperm viability and the effects of thawing temperature, the temparature of the thawing diluents on the sperm viability and acrosomal morphology of boar spermatozoa by the straw method. The result obtained were summarized as follows: 1. The sperm viablilty after thawing of the frozen semen was shown greater in the high osmolarity(392~492mOsm) than low osmolarity(300mOsm) in thawing diluent. The added levels of seminal plasma in thawing diluent did not affect the viability of frozen-thawed boar semen. 2. In terms of thawing temperature, the sperm viability was shown higher in the frozen semen thawed at 5$0^{\circ}C$ for one min. (p<0.01) than those thawed at 2$0^{\circ}C$ or 37$^{\circ}C$ for one min. The sperm viability was not significant at the diluent temparature of 2$0^{\circ}C$or 37$^{\circ}C$ after thawing: but the sperm viability was higher in thawing diluent at 2$0^{\circ}C$ than in that at 37$^{\circ}C$. However, the effects of thawing temperature and diluent solution on normal acrosomal rate were not significant. 3. Cleavage rates of oocytes fertilized with frozen semen were 46.4% and 43.3%, respectively, which were thawed at 5$0^{\circ}C$ for one min. and then diluted in mBTS medium at 2$0^{\circ}C$or 37$^{\circ}C$. To sum up, the sperm viability was shown greater at the high of thawing diluents of frozen boar semen. In terms of thawing conditions, the sperm viability was shown greater, when semen was thawed at a high temperature for a short time and then diluted at the same temperature as that in the straw.

• Korean Journal of Animal Reproduction
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• v.8 no.2
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• pp.79-86
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• 1984

This cepseiment was carried out to cerify the effect of thawing methods and preservative temperature on the sperm motility and fertility after thawing semen with plastic straws in fresh and warm water. Sperm motility in vitro stored at room temperature after thawing were conducted by the various storage hours. A field trial after thawing semen with warmed water in straws from Cheju native cows involving 4 technicians and 800 cows first (or second) services gave the following results. The thawing methods of warmed water for one minute in semen motility were considerably higher than that in iced water during 12 hours after thawing semen, however, the sperm survival index of ice-water shwed a better results according as the time passed away, but not significant differences. Preservative temperaure at 5$^{\circ}C$ (iced water) after thawing gave significantly better results than that of thawed at 3$0^{\circ}C$ (warmed water). The N R rate to 175 inseminations with semen thawed at 15-2$0^{\circ}C$ (fresh water) was 82.8%, 80.9% for 610 inseminations thawed in warm water. Conception rate ofthe semen thawed in warm water for 10-60 secs gave no significant difference among storage hours, because the semen used to be inseminated within one hour almost, but in decreased when semen thawed at the period of one minute over.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.4
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• pp.500-503
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• 2002

This study was carried out to investigate the effects of season influencing semen characteristics, frozen-thawed sperm viability and testosterone concentration in Duroc boars. There were no significant differences in the semen volume and sperm concentration of Duroc boars among spring, summer, autumn and winter. However, the pH of sperm-rich and sperm-poor fractions in autumn and winter season was higher than in spring and summer season in Duroc boars. Sperm motility and normal acrosome of raw semen in Duroc boars did not differ significantly among spring, summer, autumn and winter. However, motility and normal acrosome of frozen-thawed sperm were higher in spring season than in summer, autumn and winter. Serum testosterone concentrations in Duroc were higher in spring than summer, autumn and winter. In conclusion, when serum testosterone concentrations were higher in seasons, frozen-thawed sperm viability in Duroc boars were higher.