• 한국수정란이식학회지
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• 제12권2호
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• pp.189-194
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• 1997

The ovaries of Korean native cows or heifers were obtained from a slaughter house and kept on 28~3O˚C and transported to laboratory within 2 hrs. The follicular oocytes were collected follicles. The oocytes were matured in vitro for 24 hrs. In TCM-199 supplemented with 35 $\pi$g /ml FSH, 10 $\pi$g /ml LH, 1 $\pi$g /ml estradiol-17 and granulosa cells at 39˚C under 5% $CO_2$ in air. The caudal epididymis of Korean native bulls were obtained from a slaughter house and transported to laboratory within 30 minutes. Swim-up of collected spermatozoa and freezing sperm was layered under 2ml fertilization B. 0. medium in two tissue culture tubes and held at a 45˚C angle for 0~2 hrs. They wrer fertilized in vitro by freezing sperm treated with heparin for 24 hrs, and then the zygotes were co-cultured in vitro with bovine oviductal epithelial cells for 7 to 9 days. The follicular oocytes recovered were classified into 41.7% as grade I, 51.5% as grade II and 6.8% as graed III. The number of oocytes recovered per ovary was averaged 8.3 and they were classifed into 2.3 as grade I, 2.5 as grade II and 2.3 as grade III. The cleavage rate of matured oocytes was significantly(P

• 대한수의학회지
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• 제40권1호
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• pp.187-195
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• 2000

The ejaculates from 67 HanWoo prove bull, bred in Livestock Improvement Main center of NLCF, were used to determine the correlation between the sperm motional characteristics and the pregnancy rate of artificial insemination(AI). The motional characteristics of sperm were analysed by Computer-assisted sperm analyser(CASA), thereafter inseminated equally 1,256 heads of cow regarding to parity, age, and live weight. There were no significant difference(p>0.05) in the pregnancy rate according to year from 1996 to 1998, but the LIN, ALH, STR, BCF, MAD and WOB of sperm in the year 1997, were highest pregnancy rate, were higher than those of sperm in the year 1998, were lowest pregnancy rate(p<0.05). The semen had no significant effect on pregnancy rate according to season(p>0.05). However spring, had a little higher pregnancy rate than that of autumn, were higher than autumn in VSL, VAP, LIN, ALH, BCF, MAD and WOB, but in DNM. The pregnancy rates of spring in the year 1996 and 1997 were higher than that of autumn in the year 1998(p<0.05). The spring in the year 1997, highest in pregnancy rate, were higher than the autumn in the year 1998 in VSL, VAP, LIN, STR, BCF, MAD and WOB, but in DNM(p<0.05). There were no the motion characteristic of sperm that was significant correlate with pregnancy rate of AI as the semen were analysed before artificial insemination and those, had some degree characteristics in motility, viability and abnormality, were used to AI. However there were a tendency that the higher the VSL, VAP, ALH, LIN, STR, BCF, MAD and WOB and the lower the DNM were, the higher the pregnancy rate of AI were.

• 한국동물생명공학회지
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• 제36권2호
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• pp.91-98
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• 2021

The objective of this study was to establish a selection process for high quality sperm in bovine semen using sperm separation by magnetic activation (MACS). For this, semen from 21 Nellore bulls was collected using an artificial vagina. To guarantee the presence of pathologies in the ejaculate, animals previously declassified in four consecutive spermiogram were used. Semen was analyzed in five statuses: (1) fresh semen (fresh); (2) density gradient centrifugation (DGC), percoll column; (3) non-apoptotic fraction after separation by MACS (MAC); (4) apoptotic fraction from the separation (MACPOOR); and (5) MAC followed by DGC (MACDGC). Using a computerized analysis system (CASA), motility was measured. The sperm morphology was evaluated by phase contrast, and the supravital test was completed with eosin/nigrosin staining. For DGC, 20 × 10⁶ cells were used in a gradient of 90% and 45% percoll. MACS used 10 × 10⁶ cells with 20 μL of nanoparticles attached to annexin V, and filtered through the MiniMACS magnetic separation column. Membrane integrity was assessed with SYBR-14/IP and mitochondrial potential with JC-1 by flow cytometry. Processing sperm by MACDGC, was more effective in obtaining samples with high quality sperm, verified by the total of abnormalities in the samples: 35.04 ± 2.29%, 21.50 ± 1.47%, 17.30 ± 1.10%, 30.68 ± 1.94% and 10.50 ± 1.46%, respectively for fresh, DGC, MAC, MACPOOR, and MACDGC. The subpopulation of non-apoptotic sperm had a high number of live cells (82.65%), membrane integrity (56.60%) and mitochondrial potential (83.98%) (p < 0.05). These findings suggest that this nanotechnological method, that uses nanoparticles, is efficient in the production of high-quality semen samples for assisted reproduction procedures in cattle.

• 한국가축번식학회지
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• 제5권2호
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• pp.35-42
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• 1981

A total 2,488 ejaculates during 4 years from 80 Korean native bulls over the age from 3 to 12 years of bull herds of Artificial Breeding Center, National Livestock Federatives Cooperation were collected and analyzed to study the effects of collection year, age of bulls, month of years, collection interval and ejaculation frequency per day on semen volume, sperm concentration, total sperm per ejaculate and sperm motility. 1. Semen volume, sperm concentration, total sperm per ejaculate and sperm motility index for the all ejaculates using least squares procedure averaged 5.73ml, 9.133${\times}$108/ml, 52.527${\times}$108 and 62.81, respectively. 2. Semen volume varied significantly with collection year, collection interval and ejaculation frequency per day(P<0.01), but effects of age of bulls and month of years on semen volume were not significant. Eight to 15 days collection interval showed the highest volume, and 1st ejaculate contained 5.6% more volume than 2nd ejaculate. 3. There were significant differences among collection years, months of years and ejeculation frequencies per day except collection intervals in sperm concentration per ml(P<0.05, P<0.01). Six to 8-year-old bulls was the highest concentration. Higher sperm concentration per ml was in April to July and lower month was October and December. Sperm concentration in 2nd ejaculate was higher than in 1st ejaculate. 4. Total sperm per ejaculate affected by all environmental factors studied(P<0.05, P<0.01). Age of bulls, collection interval and ejaculation frequency per day showed the highest total sperm was 6 to 8-year-old bulls, 8 to 15 days interval and 1st ejaculate, respectively. Higher total sperm per ejaculate was in April to July and lower total sperm was in September to December. 5. In sperm motility, there were significant differences among collection years, ages of bulls and collection intervals except months of years and ejaculation frequencies (P<0.01). Higher sperm motility was in 6 to 12-year-old bulls and in 5 to 7 days collection interval.

• Applied Microscopy
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• 제41권1호
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• pp.31-35
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• 2011

본 연구는 물윗수염박쥐(Myotis daubentonii ussuriensisi)의 정자 미세구조적 특징을 알아보기 위하여 투과전자현미경(TEM)으로 관찰한 결과는 다음과 같다. 물윗수염박쥐 정자두부의 길이는 $4.5{\mu}m$, 폭은 $2.0{\mu}m$이었으며, 핵은 그 중 $4.3{\mu}m$로 정자두부의 대부분을 차지하고 있었으며, 핵의 후방부에서 전방부쪽으로 갈수록 폭이 가늘어지며 두부의 전체적인 모양은 탄환형(bull-shaped)을 취하고 있었다. 또한, 핵과 첨체는 Apical body에 의해 분리되어져 있었다. 정자경부는 basal plate, capitulum, segmented columns으로 구성되어져 있었다. 12~14개의 가는띠로 구성된 Segmented columns들의 밑부분은 중편부의 외측섬유와 연결되어져 있었다. 중편부의 미토콘드리아 초들은 나선상으로 배열되어져 있었으며, 그 수는 57개이었다. Satellite fibers는 외측섬유들 사이에 불규칙하게 관찰되어 졌으며, 중편부를 제외하고 다른 부분에서는 관찰되지 않았다. 중편부의 외측섬유는 1, 5, 6, 9번이 다른 것보다 크며, 꼬리의 축사구조는 전형적인 9+2구조이었으며, 미부의 미세소관들은 불규칙하게 배열되어져 있었다.

• 한국수정란이식학회지
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• 제25권4호
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• pp.247-250
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• 2010

• 한국수정란이식학회지
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• 제26권3호
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• pp.187-193
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• 2011

This study was carried out to establish most suitable freezing condition, to evaluate the different glycerol concentration of freezing and thawing rates on motility, viability, membrane integrity and acrosome intecrity of frozen Korean Jeju Black Bull spermatozoa, Semen was collected from a Korean Jeju Black Bull using an artificial vagina and transported to the laboratory. The semen was extended gradually 1:5 then cooled slowly for 2 hrs to 4$^{\circ}C$. The semen was diluted 1:1 with cryoprotectant extenders (3%, 5% and 7% glycerol) and equilibrated for 2 hrs at cold chamber and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 minutes, above 5 cm for 10 min and above 8 cm for 10 min. And then the frozen straw was plunged into LN$_2$. The presented straws were examined the viability and motility after thawed at 37$^{\circ}C$ water bath. The viability and membrane integrity immediately post-thawing were significantly higher in samples frozen in 7% glycerol than 3% and 5% glycerol (p<0.05). After CTC staining to assess acrosome integrity, F pattern was significantly increased, but B pattern was significantly decreased in 7% glycerol (p<0.05). Freezing distance of 5 cm from liquid nitrogen and pre-cooling for 10 min yield better survival and membrane integrity, but not significant difference. However, AR pattern according to CTC staining was significantly decreased in 3 cm for 5 min.

• Asian-Australasian Journal of Animal Sciences
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• 제16권10호
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• pp.1424-1428
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• 2003

Present investigation was conducted to determine the post-thaw sperm motility and acrosomal damage of filtered and non-filtered frozen semen of Murrah buffalo bulls. Twenty semen ejaculates (from four Murrah buffalo bulls collected at weekly interval) were diluted in Tris egg yolk glycerol extender and divided into two parts. One was filtered through sephadex G-100 column and the other portion was kept as such (non-filtered). Both fractions were frozen in liquid nitrogen ($-196^{\circ}C$) by the standard method developed in the laboratory. After 24 h of freezing, non-filtered and filtered semen samples were thawed at $37^{\circ}C$ for 1 min. These samples were incubated at $37^{\circ}C$ in a water both. The different seminal characteristics i.e. percent progressive sperm motility, live and abnormal spermatozoa and spermatozoa with damaged acrosome were assessed at hourly interval till they remained motile. The filtered frozen and thawed semen showed significantly (p<0.05) high sperm viability and acrosomal integrity as compared to non-filtered semen.

• Animal Bioscience
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• 제34권2호
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• pp.198-204
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• 2021

Objective: The aim of this study was to ascertain the effects of adding green tea extract (GTE) to skim milk-egg yolk (SM-EY) extender on both the quality of post-thawed bull semen and the pregnancy rates of the recipient cows. Methods: Twelve ejaculates from four Simmental bulls, aged 3 to 5 years and weighing 900 to 950 kg, were diluted SM-EY extender, added with 0, 0.05, 0.1, and 0.15 mg GTE/100 mL extender and then frozen. After four weeks storage in liquid nitrogen, the sperm were thawed and evaluated for viability, motility, intact plasma membrane (IPM), and DNA fragmentation. Meanwhile, the estrus cycles of 48 recipient cows were synchronized by intramuscular administration of a single injection of 5 mg prostaglandin F2α. Estrus cows were divided into four equal groups and inseminated artificially 18 to 20 h after the onset of estrus by using semen from each extender group. Pregnancy was diagnosed by measuring serum progesterone levels at 21 days, followed by transrectal palpation 90 days after insemination. Results: The findings revealed that adding 0.1 mg of GTE/100 mL extender produced the highest percentages of sperm viability (70.67%±1.75%), motility (69.17%±1.47%), and IPM (69.23%±1.21%) and the lowest percentage of DNA fragmentation (3.00%±0.50%). The pregnancy diagnosis revealed that all cows (36/36) inseminated using frozen semen in GTE addition extender were pregnant (pregnancy rate 100%), whereas the pregnancy rate of the control group was 83.33% (10/12). Conclusion: It may be concluded that 0.1 mg GTE/100 mL extender yields the best quality of spermatozoa and that all variants doses of GTE in extender produce a higher pregnancy rate among recipient cows.

• Journal of Animal Science and Technology
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• 제44권6호
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• pp.693-700
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• 2002

한우 종모우에 있어 소 성장호르몬 유전자의 출현빈도를 알아보고 소 성장호르몬 다형과 정액성상과의 관계를 살펴보기 위하여 실시하였다. 한우 종모우 109두의 소 성장호르몬 유전자를 Alu I 제한효소로 처리하여 PCR-RFLP로 분석하였다. Leucine(Leu)과 valine(Val) 유전자의 출현빈도는 각각 0.88과 0.12이었다. 소 성장호르몬 VV 유전자형을 지닌 한우 종모우가 다른 유전자형을 지닌 종모우보다 정액성상(정액량, 정자농도, 총정자수)이 떨어지는 경향을 보였지만 소 성장호르몬의 유전자형이 정액성상에 유의적인 영향은 미치지 못하였다. 채취순번에 따른 영향에 있어서도 VV 유전자형을 지닌 종모우가 다른 유전자형을 지닌 종모우보다 정액성상이 떨어지는 경향을 보였지만 유의적인 차이는 나타내지는 못하였으며 다만 년도에 따른 영향에 있어 1998년도 VV 유전자형을 지닌 종모우가 다른 유전자형을 지닌 종모우보다 총정자수에 있어 유의적으로 적게 나타났다(P<0.05). 전체적으로 소 성장호르몬 유전자의 VV 유전자형을 지닌 종모우의 정액성상이 다른 유전자를 지닌 종모우의 정액성상보다 낮은 경향을 보였지만, 조사한 109두의 한우 종모우 중 VV 유전자형을 지닌 종모우가 1두로 분석되어 이 종모우의 정액성상만을 이용하여 소 성장호르몬 유전자다형이 정액성상에 미치는 영향을 살펴보았기에 종모우 선발시 기초자료로 이용하기에는 추가적인 시험이 필요할 것으로 사료된다.