• Korean Journal of Animal Reproduction
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• v.3 no.2
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• pp.57-63
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• 1979

In this study, pooled whole semen and diluted semen with skim milk lactose solution and yolk skim milk lactose solution were inseminated at 6 and 7 days interval during 90 days. Egg production, fertility and hatchability were investigated. The results obtained from this study are summarized as following: 1. The average fertility of 57.8% for whole semen was clearly higher than that of 35.2% for diluted semen during 90 days insemination trial. 2. The highest fertility was a, pp.ared from 35 to 99 days after insemination for the whole semen, but from 28 to 60 days for the diluted semen during 90 days insemination trial. 3. In case of 7 days insemination interval, highest fertility of 86.6 and 70.0% for the whole semen and the diluted semen was a, pp.ared on 2 days after insemination and thereafter the fertility was gradually decreased according to passage of insemination. The lowest fertility of 35.0 and 0.0% for the whole semen and diluted semen was a, pp.ared on 1 day after insemination. 4. In case of 6 days insemination interval, highest fertility of 80.0 and 55.8% for the whole semen and the diluted semen was also a, pp.ared on 2 days after insemination and thereafter the fertility was slowly decreased according to passage of insemination. However, lowest fertility of 25.0 and 20.0% for the whole semen and the diluted semen was a, pp.ared on 0 day after insemination. 5 It suggests that there was no difference in fertility between the skim milk lactose and the yolk skim milk lactose dilutors. 6. In case of whole semen, average fertility of 7 days insemnaition interval was a, pp.rently lower than that of 6 days, however there was no difference in fertility between 6 and 7 days insemination interval. 7. Insemination interval of 6 and 7 days and passage day after insemination did not alter egg production and hatchability of fertilized egg production and hatchability of fertilized egg in both whole and diluted semen.

• Korean Journal of Animal Reproduction
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• v.21 no.1
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• pp.25-30
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• 1997

This study was done to find out the preservation possibility of liquid boar semen at variabel room temperature of 9 to 16$^{\circ}C$. The percentages of sperm motility and NAR acrosome were highest in B tschwiler extender compared to B tschwiler＋Hepes, Andro＋Hepes and Andro extenders. The extenders with Hepes buffer showed detrimental effect for preservation of liquid boar semen. The pH of ejaculated sperm-rich fraction was 7.5. The pH of B tschwiler＋Hepes, B tschwiler, Andro＋Hepes and Andro extenders was 6.9, 7.5, 7.1 and 8.1, respectively. The pH of liquid boar semen with B tschwiler＋Hepes, B tschwiler, Andro＋Hepes and Andro extenders was 6.6, 6.9, 6.7 and 6.9 at 1st day of storage, and 5.5, 5.7, 5.6 and 5.8 at 7th day of storage, respectively. Gilts and sows were inseminated twice with liquid boar semen stored at 9~16$^{\circ}C$ in B tschwiler extender for 3~4 days. Farrowing rate, litter size and average pig weight at birth between AI and natural service did not differ significantly in gilt and sow, respectively. However, sow showed higher farrowing rate and litter size compared to gilt both in AI and in natural service. As a result of this study, we found out that liquid boar semen can be stored for 5~7 days at uncontrolled room temperature of 9~16$^{\circ}C$ in B tschwiler extender.

• Korean Journal of Animal Reproduction
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• v.21 no.1
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• pp.19-23
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• 1997

This present study was carried out to examine the effect of maturation media and liquid boar semen on in vitro maturation and feritilization of pig oocytes. The results obtained were as follows : When the oocytes were cultured for 36∼42 hours in mTCM-199, Waymouth MB 725/1 and mTLP-PVA medium, the maturation rates were 90%, 92% and 88%, respectively. The sperm penetration rates of pig oocyte matured in vitro were 87%(mTCM-199), 90%(Waymouth MB 725/1) and 86%(mTLP-PVA), respectively. The rates of nuclear maturation and fertilization of pig oocytes among three different media did not differ. However, the rate of male pronucleus formation of pig oocytes was significantly higher in pig oocytes matured in Waymouth MB 725/1(91%) than oocytes matured in mTCM-199(66%) and mTLP-PVA(62%) medium (P<0.05). When the collected sperm-rich fraction without diluent was used fro in vitro fertilization in mTCM-199 fertilization medium, the fertilization rate was 87.9%. However, when the liquid boar semen diluted with B tschwiler diluent was used at day 3 and 5 after dilution, the fertilization rate was 40.8% and 0.0%, respectively.

• Analytical Science and Technology
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• v.12 no.3
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• pp.260-264
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• 1999

A simple and rapid procedure, called FoLT-PCR(Formamide Low Temperature-Polymerase Chain Reaction) was applied to amplifying DNA directly from various forensic biological evidences including human blood, saliva, hair root, or semen without any DNA preparative steps. We added washing step with non-ionic detergent, 1% Triton X-100, and used Taq DNA polymerase instead of Tth DNA polymerase to amplify 3 STR loci and gender allele simultaneouly. Optimal concentration of formamide and annealing temperature were determined empirically to 8%(v/v), and $48^{\circ}C$ respectively. We also compared this method with standard PCR.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.239-246
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• 1992

Capacitated and acrosome~reacted spermatozoa were microinjected into the perivitelline space of bovine oocytes matured in vitro. Oocytes obtained from the ovaries of slaughtered heifers and cows were cultured in vitro in the TCM-199 supplemented with 20% FCS for 24 hr at 39$^{\circ}C$ under an atmosphere of 5% CO$_2$ 8% O$_2$. Fresh or frozen spermatozoa were incubated for 2 hr at 39°C under an atmos-phere of 5% CO$_2$, 8% O$_2$ in Ham's F-lO medium containing 0.75% BSA for capacitation, and kept for 30 min in culture medium containing 12 mM of dbcGMP and lOmM of immidazol for acrosome resction. One motile spermatozoon was injected into the perivitelline space of each oocyte. The 2nd polar body and the pronuclei were observed in 9.5% and 5.4% of oocytes, respectively. The rate of cleavage of oocyte over 2-cell stage was 4.1%(10 of 242), These results indicate that the microinjection may be a useful technique to study sperm-oocyte interaction.

• 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.

• Journal of Embryo Transfer
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• v.14 no.2
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• pp.99-106
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• 1999

핵 이식에 공여되는 수핵난자의 효과적인 동결보존을 위하여 한우 성숙난자를 1.0 M dimethylsulfoxide(DMSO) 또는 1.0 M glycerol이 함유된 동결보호제를 이용하여 처리하거나, 동결보호제 처리 후 완만동결법을 이용한 동결융핼르 시행하여 상기 실험처리로 야기되는 투명대 경화현상을 관찰하였다. 도축장 유래의 난소에서 미성숙난자를 채취한 후 10% 소 태아혈청을 함유한 TCM-199을 이용하여 22∼24 시간 동안 체외성숙배양을 이해하였다. 배양후 작출된 성숙난자를 각각의 동결보호제로 처리, 혹은 처리 후 동격융해한 후 protease를 이용하여 투명 대의 경화현상 발생의 빈도를 조사하였다. 또한 동격란을 동결정액을 이용한 체외수정에 공여한 후 정자 침입농도능을 조사하였다. 동결보호제로 처리한 난자에 있어서 보호제의 종류와 관계없이 투명대 경화현상이 유의적 (P<0.05) 으로 증가하였으나 이후의 동결융해 처리에 의한 추가적인 경화현상의 발생은 증가하지는 않았다. 또한 투명대 경화현상의 발생양상을 동결보호제 처리 후 10분 간격으로 측정한 결과 DMSO의 경우 처리후 10분, glycerol의 경우 처리 후 20분 후부터 유의적으로 차를 발견할 수 없었으며, 수정율 및 난자 1개당 침입한 정자의 수는 동결란에서 유의적으로 증가하지 않았다. 본 연구의 결과 동결난자의 투명대 경화현상은 동결보호제 처리과정에서 이미 일어나지만, 이러한 투명대 경화현상이 난자의 동결보존 후 수정능에는 현자한 영향을 미치지 않는다는 사실이 규명되었다.

• Journal of Veterinary Clinics
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• v.21 no.4
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• pp.363-368
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• 2004

To investigate the effects of seeding during freezing procedure on post-thaw viability, motility, and acrosome integrity of boar spermatozoa, semen from 5 Yorkshire boars were collected for this experiment. Raw semen were diluted with Merck I, subsequently added with cooling diluent containing lactose and egg yolk and with freezing diluent containing glycerol. The diluted semen were frozen on the rack in the styrofoam box filled with liquid nitrogen at the distance of 5 cm or I cm above LN2 level. Seeding was performed to only a group of straws frozen at 5 cm away on the surface of LN2. The frozen semen were thawed in $50^{\circ}C$C water and the viability and local motility were analyzed by sperm analysis imaging system. A part of thawed semen was taken for the examination of morphology of apical ridge of the acrosome to compare with the effect of seeding between the seeding-treated and non treated groups. I. Post-thaw viability was considerably higher in seeding-treated sperm than non-seeding group (p<0.01), however, no difference of local motility was obtained among the groups. 2. At three hours after thawing, viability was also higher in seeding-treated group than non-treated group (p<0.05), along with no difference of motility among the groups. 3. Higher normal acrosome integrity was obtained in the seeding-treated sperm than non-treated groups (p<0.01). 4. Between non-seeded groups, higher normal acrosome integrity was obtained in the sperm group frozen at 5cm upper on the surface of LN2 than that frozen at 1cm away (p<0.01). These results indicated that seeding treatment during freezing boar spermatozoa was beneficial to post-thaw viability and normal acrosome integrity.

• Clinical and Experimental Reproductive Medicine
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• v.3 no.2
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• pp.35-42
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• 1976

The human semen ejaculated in a form of liquid state, coagulates immediately after ejaculation, and then liquefies again. However, the mechanisms of neither coagulation and liquefaction of semen have not been explained clearly so far, and very limited numbers of report are available, although the spermatology and andrology made rapid progress. This clinical study has been undertaken to investigate the liquefaction phenomena and practicability of the results might be applied to fertility and infertility problems. As a preliminary study, in this report the liquefaction time of various semen groups is measured and analysed. The following results are obtained: 1. An average liquefaction time of semen of a total of 60 subjects: 25 minutes. 2. An average liquefaction time of semen according to sperm count: 1) Normospermia group (20 cases): 34 minutes. 2) Oligospermia group (20 cases): 21 minutes. 3) Azoospermia group (20 cases): 20 minutes. 3. An average liquefaction time of semen according to abstinence period: 1) Less than 3 days group (30 cases): 22 minutes. 2) More then 5 days group (30 cases): 28 minutes. In conclusion: 1. The liquefaction time of semen of the normospermia group is longer than oligospermia group or azoosermia group. 2. The liquefaction time of semen may not be greatly influenced by the various factors such as abstinence period, semen volume, semen pH, age of the subjects and so on. 3. In routine semen analyses, it is recommended to begin the analysis at least 25 minutes after the ejaculation. 4. Further studies are required in conjunction with practical application of liquefaction mechanism in infertility and fertility control.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.311-318
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• 1996

Mycoplasmas have long been suspected of contributing to involuntary infertility in couples. However considerable disagreement exits concerning the role of genital mycoplasma infection in human infertility. Several investigators have noted abnormalities in the semen analysis of men with positive mycoplasma cultures, and early epidemiologic studies indicated that Ureaplasma urealyticum was linked to human reproductive failure on the basis of higher frequencies of isolation from infertile versus fertile couples and successful pregnancies in infertile couples after doxycycline therapy. However, subsequent investigators have questioned these findings because there are many studies in which treatment for mycoplasma in the male or female did not demonstrate an improved pregnancy rate, and semen samples from unexplained infertile men containing ureaplasmas have not revealed poorer motility, fewer spermatozoa and more aberrant forms. The objective of this study were to investigate the incidence rate of mycoplasma in semen and to investigate whether the presence of mycoplasma in semen makes significant difference to the semen volume, sperm motility and sperm counts. The results were that the rate of isolation of mycoplasma species was 70.3%. Semen volume is $2.84{\pm}1.01ml$ for culture negative and $3.15{\pm}1.42ml$ for culture positive group. Sperm motility is $46.23{\pm}15.80%$ for culture negative and $50.09{\pm}15.69%$ for culture positive group, and sperm count is $95.47{\pm}47.14({\times}(P)10^6/ml)$ for culture negative and $86.73{\pm}47.59({\times}10^6/ml)$ for culture positive group. In conclusion, we suggest that the presence of mycoplasma in semen makes no significant differences to the sperm parameters.