• Reproductive and Developmental Biology
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• 제29권4호
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• pp.235-239
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• 2005

This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine $\beta-casein/human$ lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in $50\;{\mu}L$ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was $20.9\%$ (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 $(8.8\%)$ were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.

• 한국수정란이식학회지
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• 제26권1호
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• pp.9-12
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• 2011

This study was undertaken to determine whether the presence of fertility-associated antigen (FAA) in semen would influence semen characteristics and conception rate of artificial insemination in Hanwoo. The response to FAA of 36 heads of proven bull, 7 heads of young bull, and 27 heads of performance-tested bull was that one proven bull was FAA-negative and the others were FAA-positive, therefore FAA-negative bull was 1.4%. FAA-negative bull was lower in first and second semen concentrations than those of FAA-positive bull in 5,301 semen of 21 heads of proven bull, then FAA-negative bull was fewer as 11.5% in total sperm counts. The estrus of 22 heads was 70d-nonreturned in 36 cows first inseminated with frozen semen of FAA-negative bull, but that of 249 heads in 378 cows first inseminated with frozen semen of FAA-positive bull. Each conception rate was 61.1% and 65.9%, respectively. The difference of conception rates was 4.8%. These results indicate that the response of FAA to semen were influenced semen characteristics and conception rate of artificial insemination, but further investigations are needed to confirm the results.

• Clinical and Experimental Reproductive Medicine
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• 제11권2호
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• pp.69-76
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• 1984

In order to obtain high proportion of Y sperm the semen was laid over 6%,10% and 20% bovine serum albumin. Separated sperm were stained with quinacrine mustard in order to see F - body which could be seen in human Y sperm. But we could'nt find F-body in the bull sperm. So sperm were compared with size of sperm head. As a result of observation separated sperm was small in size of length and width of sperm head as compared with control sperm. So it was found that the proportion of Y sperm showed a marked increase in separated layer. Then the higher albumin density was, the higher the proportion of Y sperm which had smaller head and faster motility was. But the higher albumin density was, the lower the recovery rate of sperm was. So it was hard to separate Y sperm in oligospermia.

• 한국동물위생학회지
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• 제40권3호
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• pp.201-208
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• 2017

소에 있어서 BVDV는 반추류에서 중대한 감염을 야기하여 번식장애, 설사 및 유량감소를 야기하여 농가에 막대한 피해를 야기하고 있다. 특히 지속감염우의 확인과 도태는 농가 내 감염원을 제거하는 중요한 일이나 아직까지 PI개체의 바이러스의 감염과 전파에 관한 연구는 미진한 것으로 추정된다. 본 연구에서는 도입된 수컷에서 PI를 확인하였으며 전혈 검사를 실시 하면, PI 혈액 성상에서 림프구가 낮게 관찰되며 전체 WBC의 수는 정상범위에 속하나 낮게 나타나는 것을 알 수 있었다. PI 수컷에서 생산된 정액은 정자 수가 매우 낮으며, 신선 정액의 생존성도 불량한 것으로 조사되었다. 정자 기형율 또한 증가되었으며 특히 공포와 중편부의 소적을 가진 기형정자의 비율이 높았다. PI수컷은 나이가 들수록 정액 성상이 불량하여 불임화되었고 이는 BVDV가 정소의 정자 생산능력을 낮추어 사출된 정액내 정자수 감소 현상을 야기한다는 것을 알 수 있었다. 그럼에도 불구하고 확인된 PI 개체에 의한 전체군의 감염 현상은 관찰할 수 없었는데, 이는 적절한 백신 프로그램에 의하여 PI에 의한 간접적인 전파의 위험성이 낮다고 판단된다.

• 한국수정란이식학회지
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• 제21권2호
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• pp.95-100
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• 2006

한우 종모우에 있어 정액 채취시 승가 조건(의빈우 혹은 의빈대)이 정액성상에 미치는 영향을 구명하기 위하여 의빈우 승가 종모우 21두, 의빈대승가 종모우 9두 등 보증 종모우 30두를 공시하여 연구하였다. 정액은 $3{\sim}6$일 간격으로 채취하였으며 채취 당일 1시간 간격으로 총 2회 정액 채취하여 6개월간 정액성상을 조사하였다. 의빈대 승가 종모우가 의빈우 승가 종모우보다 1차 및 2차 정자농도에 있어 유의적으로 높아(p<0.05) 1차 및 2차 총 정자수에서도 많았으며 특히 l차 총 정자수는 유의적으로 많게 나타났다(p<0.05). 종모우 승 가 조건(의빈대, 의빈우)에 상관없이 5일 간격으로 정액 채취시가 정액량이 많아(p<0.05) 총 정자수가 많았으며 의빈대를 이용하여 5일 간격으로 정액 채취시가 총 정자수가 다른 모든 처리군보다 많았다. 의빈대 승가 종모우는 $4{\sim}5$세에서 총 정자 수가 높게 나타났으며 의빈우 승가 종모우는 $6{\sim}7$세에서 총 정자수가 높게 나타났다(p<0.05). 이러한 결과로 미루어 볼 때 한우 종모우에서 의빈대를 이용하여 정액을 채취하는 것이 의빈우를 이용 할 때 보다 정액 생산성이 좋을 것으로 사료된다.

• 한국가축번식학회지
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• 제26권3호
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• pp.275-289
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• 2002

본 연구는 종모우의 정자수정능력 평가방법의 개발과 정자 기능 및 성상 변화에 영향을 미치는 요인을 알아보기 위하여 시행하였다. 동결-융해된 종모우 정액을 대상으로 정자의 운동성과 정자의 형태를 분석하였고, 정자의 기능 검 사 항목으로서 체외수정(IVF), HOST, Ca-ionophore에 의한 첨체반응율, 정자의 ROS 측정을 위한 luminol, lucigenin-dependent chemiluminescence, LPO 분석을 위한 malondialdehyde의 측정 및 TUNEL (terminal deoxynucleotidyl transferase(TdT) dUTP nick end labelling) 기법을 이용한 정자의 DNA fragmentation를 측정하였으며 이들 각각의 조사 항목들의 분석치들과 체외수정율 및 배발생율과의 상관관계를 조사하여 다음과 같은 결과를 얻었다. 1. 고수정군과 저수정군의 체외수정율과 배반포 발생율의 평균은 각각 64.4%와 34.3%, 18.50%와 6.2%였으며 두 군간에 통계학적으로 유의한 차이를 보였다(P<0.05). 고수정군과 저수정군의 정자운동성과 첨체반응률은 각각 평균79.0 %와 66.2%, 40.7%와 22.9%로 두 군간에 통계학적으로 유의한 차이를 보였으나(P<0.05), 정상형태 정자의 비율과 HOST는 각각 평균 94.6%와 92.7%, 69.4%와 59.8%로 두 군간 유의한 차이를 보이지 않았다. 2. Luminol dependent chemiluminescence, LPO 및 DNA fragmentation의 평균은 고수정군과 저수정군에 있어서 각각 6.4와 6.5, 2.Onmol와 3.Inmol 및 2.6%와 7.4%로 두 군간 통계학적으로 유의한 차이를 보였으나(P<0.05), lucigenin dependent chemiluminescence는 4.7와 4.6로 두 군간 유의한 차이를 보이지 않았다. 3. 체외 수정율은 정자의 운동성 및 첨체반응율과 통계학적으로 유의한 정(positive)의 상관관계(r=0.87, p<0.01; r=0.81, p<0.05)를 나타내었으며, luminol dependent chemiluminescence, lipid peroxldation 및 DNA fragmentation과는 통계학적으로 유의한 부(negative)의 상관관계 (r= -0.81, p<0.05; r: -0.74, p<0.05; r : 0.81, p<0.05)를 나타내었다. 그러나 체외수정율은 정상형태 정자의 비율, HOST 및 lucigenin dependent chemiluminescence와는 유의한 상관 관계를 나타내지 않았다. 4. 배반포 발생율은 첨체반응율과 통계학적으로 유의한 정의 상관관계(r=0.71, p<0.05)를 나타내었으며, luminol dependent chemiluminescence, lipid peroxidation 및 DNA fragmentation과는 통계학적으로 유의한 부의 상관관계(r= -0.71, p<0.05; r= -0.89, p<0.01; r= -0.71, P<0.05)를 나타내었다. 배반포 발생율은 정자의 운동성, 정상형태 정자의 비율 및 HOST, lucigenin dependent chemilumihescence와는 유의한 상관관계를 나타내지 않았다. 이상의 결과를 종합해 보면 정액질의 저하에 ROS의 영향이 밀접히 연관되어 있음을 알 수 있으며, 또한 본 연구에서 적용된 기법들은 정액질의 평가 및 정자 수정능력 향상을 위한 기술개발에 있어서 유용한 평가 방법으로 이용될 수 있을 것으로 사료된다.

• Animal Bioscience
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• 제37권2호
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• pp.203-209
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• 2024

Objective: This study aimed to assess the impact of the dilution ratio of Tris diluent, storage at 0℃, and long-distance transportation on the spermatozoa of Simmental cattle. It also validated the feasibility of the regional distribution of fresh semen. Methods: In experiment 1, semen was diluted at four dilution ratios (1:6, 1:9, 1:12, and 1:15) to determine the optimal dilution ratio of Tris diluent. In experiment 2, we assessed sperm viability, progressive motility (objectively assessed by computer-assisted sperm analyzer), and acrosome intactness in Tris dilutions kept at constant 0℃ for 1, 3, 6, 9, and 12 days. We compared them to Tianshan livestock dilutions (Commercial diluent). In experiment 3, semen was diluted using Tris diluent, and sperm quality was measured before and after long-distance transport. Artificial insemination of 177 Simmental heifers compared to 156 using Tianshan Livestock dilution. Results: The outcomes demonstrated that 1:9 was the ideal Tris diluent dilution ratio. The sperm viability, Progressive Motility, and acrosome integrity of both Tris and Tianshan dilutions preserved at 0℃ gradually decreased over time. sperm viability was above 50% for both dilutions on d 9, with a flat rate of decline. The decrease in acrosome integrity rate was faster for Tianshan livestock dilutions than for Tris dilutions when stored at 0℃ for 1 to 6 days. There was no significant difference (p>0.05) in sperm viability between semen preserved in Tris diluent after long-distance transportation and semen preserved in resting condition. The conception rates for Tris dilution and Tianshan livestock dilution were 49.15% and 46.15% respectively, with no significant difference (p>0.05). Conclusion: This shows that Tris diluent is a good long-term protectant. It has been observed that fresh semen can be successfully preserved for long-distance transport when stored under 0℃ conditions. Additionally, it is feasible to distribute semen regionally.

• 한국가축번식학회지
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• 제12권1호
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• pp.24-30
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• 1988

These experiments were carried out to develop new techniques for In Vitro separatin of X-and Y-bearing spermatozoa. The bull semen was applied to the various Gel-Columns filled with swellen Sephadex G-50 Fine and then elutriated wtih Locke solution (elutriation rate; 1ml/3-4min., 1ml/1-2min.). Elutriated solution was fractionated into 1ml by automatic Fraction Collector and spermatozoa included in each fraction were subjected to the estimation of viability and recovery rate, and to B-body test. The results obtained in these experiments were summarized as follows: 1. When the column size and the elutriation rate were adjusted to 15$\times$1.6cm and 1ml/3-4min., respectively, the highest sperm concentration was obtained from the 8th to the 12th fraction. 2. As a trend, the viability of spermatozoa was improved by chromatography, and the degree of improvement ranged 5 to 10 percentage. 3. The average recovery rate of spermatozoa applied to column was 73.2 percentage and ranged 52.6 to 81.3 percentage. 4. The lowest rate of B-body bearing spermatozoa following chromatography was obtained when the column size and the elutriation rate were adjusted to 15$\times$0.8cm and 1ml/1-2min., respectively.

• 한국수정란이식학회지
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• 제15권1호
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• pp.95-102
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• 2000

Techniques for manipulation of spermatozoa and oocytes have been widely used for in vitro production(IVP) of Hanwoo. This study was conducted to examine the effects of theophylline and heparin on frozen-thawed Hanwoo sperm for enhancing the efficiency of IVP technique. Oocytes were inseminated with forzen bull semen treated with either theophylline or heparin for examining the effect of each substance on fertilization and subsequent development. More (P<0.05) oocytes formed pronucleus and develop to the morula and blastocyst stages after inseminated with sperm treated with heparin than after inseminated with sperm treated with theophylline. The pregnancy rate after embryo transfer was higher after heparin treatment than after theophylline treatment, but did not differ significantly. There was no significant difference of offspring delivery between two groups. In conculsion, theophylline and heparin can be used for enhancing the efficiency of IVP system for Hanwoo. Considering characteristics of these substance, theophylline may be useful in the artificial insemination system, which requires vigorous sperm motility. While, heparin supporting sperm viability in vitro can be effectively used for improving in vitro-fertilization system.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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• pp.56-56
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• 2001

This study was carried out to investigate on the improvement of fertilizing and developing ability of in vitro matured oocytes from individuals of bulls, sperm type, pretreatment of sperm or oocytes obtained by intracytoplasmic sperm injection(ICSI). 1. The male pronuclear formation and developmental rates of oocytes obtained by ICSI treated individual of bulls were 73.9%-87.0% and 33.3%-60.9%, respectively. 2. The male pronuclear formation and developmental rates of oocytes obtained by ICSI treated fresh and frozen sperm, tail-cutting and tail-scoring sperm were 82.0%, 78.0%, 42.2%, 51.1% and 56.0%, 42.0%, 17.8%, 22.2% respectively. and these values of fresh sperm injection were higher than that of frozen sperm, tail-cutting and tail-scoring. 3. The male pronuclear formation and developmental rates of oocytes obtained by sperm pretreated heparin, BFF(bovine follicula fluid), His, Ca Ionophore(Ⅰ) and Ⅰ ＋ caffeine methods were 66.7%-82.2% and 33.3%-60.6%, respectively. and these values of treatment of Ⅰ＋ caffeine were higher than that of other methods. 4. The male pronuclear formation and developmental rates of oocytes obtained by ICSI treated with or without zona pellucida were 80.0%, 72.0% and 46.0%, 36.0%, respectively.