• 한국수정란이식학회지
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• 제24권4호
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• pp.271-274
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• 2009

To establish a protocol of estrus induction and synchronization in European mouflon, we performed artificial insemination using frozen-thawed semen and exogenous hormones. CIDR was inserted into vaginas of four mouflons for 16 days. A day before removal of CIDR, PG 600 was injected intramuscularly. $PGF_2{\alpha}$ was injected when removing CIDR. Artificial insemination was cervically conducted with injecting LHRH 48 hours after CIDR withdrawal. Even though no pregnancy was confirmed, estrous signs were notified like open cervix, congestion of vaginal wall and discharge of cervical mucus. Further research in the wild sheep would be needed for development of artificial breeding methods and advancing sustainability of domestic zoos.

• 한국가축번식학회지
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• 제17권4호
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• pp.271-278
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• 1994

The aim of this study was to compare the two culture systems 1) co-culture with cumulus cells and 2) chemically defined medium supplemented with amino acids (CR1aa) and fetal calf serum (FCS) of in vitro produced bovine embryos from follicular oocytes in vitro. Bovine follicular oocytes were collected from ovaries of slaughtered cows and matured in TCM199 supplemented with 10% FCS and hormones (1$\mu\textrm{g}$/ml FSH-P and 1$\mu\textrm{g}$/ml oestradiol-17$\beta$)24 hours at 39$^{\circ}C$ under 5% CO2 in air. The capacitation of spermatozoa from ejaculated or frozen bull semen was induced by centrifugation through Percoll density gradient (45%, 90%). Then capacitated spermatozoa (1$\times$106/ml) were inseminated into 50${mu}ell$ droplet containing matured follicular oocytes and incubated for 40~42 hours. Cleaved embryos of 2~4cell stage were transferred to the co-culture with cumulus cells and/or CR1aa medium supplemented with FCS. In semen source, the developmental rates to the blastocyst and the hatched blastocyst stages were higher in ejaculated semen(27.6% and 14.9%) than those of frozen-thawed semen(18.3% and 11.8%), respectively. In two culture systems, the proportions of embryonic development upto the blastocysts and the hatched blastocysts were higher of CR1aa medium (22.1% and 12.1%) than those of cumulus cell co-culture (16.8% and 5.1%), respectively. The number of cells in exapnded blastocysts was slightly higher in cumulus cells co-culture (122.6$\pm$8.5) than that in CR1aa medium (117.9$\pm$5.9). The present results indicated that the early development of in vitro produced bovine embryos can be maintained efficiently in CR1aa medium as well as in co-culture with cumulus cells.

• Animal Bioscience
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• 제36권10호
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• pp.1530-1535
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• 2023

Objective: Semen cryopreservation result in decreased sperm parameters and fertilization ability. Sericin exhibits antioxidant activity by reducing lipid peroxidation resulting from free radicals, which can potentially improve cryopreservation outcomes. The present study aimed to examine the efficacy of various sericin concentrations supplemented with a rooster semen-freezing extender on post-thaw semen quality and fertilizing ability of sperm after cryopreservation. Methods: Semen samples were collected from 40 roosters (5 reps), then were pooled, and divided into four groups by the levels of sericin supplementation (0%, 0.25%, 0.50%, and 0.75%) in a freezing extender. Semen suspensions were loaded in medium straw (0.5 mL) and cryopreserved with the traditional liquid nitrogen vapor method. Post-thawed semen was evaluated for sperm motility, sperm viability, and lipid peroxidation. Also, the fertility test was determined. Results: The results showed that supplementation of the freezing extender with 0.50% to 0.75% sericin resulted in greater total motility and progressive motility and lower malondialdehyde levels than the other groups after cryopreservation (p<0.05). However, the viability of 0.75% decreased compared with the value of 0.50% sericin supplementation (p<0.05). Moreover, the fertility and hatchability of total eggs were significantly higher in the 0.50% sericin group than in the other groups (p<0.05). Conclusion: In conclusion, 0.50% sericin is recommended as an alternative component of the freezing extender to improve cryopreserved rooster semen.

• 한국수정란이식학회지
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• 제29권2호
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• pp.163-169
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• 2014

The objective of this study was to evaluate the characteristics of Korean Native Cattle sperm frozen-thawed with L-cysteine and/or catalase. The semen from bulls was collected by the artificial vagina method, and Triladyl containing 20% egg-yolk and/or L-cysteine (L), catalase (C) and L-cysteine + catalase was added to the diluted semen for cryopreservation. The results showed that sperm viability was significantly higher in the L-cysteine + catalase ($69.49{\pm}3.16%$) group than in the control ($60.5{\pm}3.94%$) group (p<0.05). Acrosome damage was significantly lower in the L-cysteine ($17.12{\pm}1.08%$) group than in the control ($21.46{\pm}1.14%$), catalase ($20.54{\pm}0.76%$), and L-cysteine + catalase ($19.29{\pm}0.65%$) groups (p<0.05). In addition, the level of intact mitochondria in the spermatozoa was significantly higher in the L-cysteine ($58.65{\pm}1.39%$) group than in the control ($50.63{\pm}2.37%$) group (p<0.05). The hydrogen peroxide level in the frozen-thawed sperm was significantly lower in the L-cysteine ($3.74{\pm}1.66%$), catalase ($4.65{\pm}1.87%$), and L-cysteine + catalase ($8.11{\pm}2.15%$) groups than in the control ($13.22{\pm}1.6%$) group (p<0.05). The glutathione level was significantly higher in the L-cysteine ($1.33{\pm}0.03%$) group than in the control ($1.08{\pm}0.06%$), catalase ($1.05{\pm}0.02%$) and L-cysteine + catalase ($1.11{\pm}0.03%$) groups (p<0.05). In conclusion, L-cysteine and catalase could protect the membrane of Korean Native Cattle sperm from damage during sperm cryopreservation. Especially, L-cysteine was more effective for keeping acrosomes and mitochondria intactness during sperm cryopreservation.

• Reproductive and Developmental Biology
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• 제40권3호
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• pp.27-31
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• 2016

The aim of this study was to evaluate effect of ${\alpha}$-linolenic acid (ALA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved in 20% egg yolk freezing extender containing ALA (0, 3, 5, and 10 ng/mL) with 0.05% ethanol. The frozen-boar spermatozoa were thawed at $37.5^{\circ}C$ for 45 sec in water-bath. The spermatozoa samples were evaluated the plasma membrane integrity, acrosome reaction, and mitochondrial integrity using flow cytometry. In results, population of live sperm with intact plasma membrane was significantly higher in control and 3 ng/mL ALA treatment group than ethanol group (p<0.05). In contract, dying sperms were higher in ethanol group than 3 ng/mL ALA treatment (p<0.05). Acrosomal membrane damage in all sperm population was reduced in 3 ng/mL ALA groups compared with ethanol treatment (p<0.05). However, acrosome damage in live sperm population was no significant difference among the all treatment groups. Mitochondrial integrity was not influenced by ALA treatments in both of live and all sperm population. In conclusion, this results show that supplement of ALA during the cryopreservation process could reduce the membrane damages including plasma and acrosomal membrane, whereas ALA did not influence to mitochondria in boar spermatozoa. Therefore, these results suggest that ALA can protect against the membrane damage derived cryo-stress, and cryopreservation efficiency of boar semen would be improved by use of ALA.

• Journal of Animal Science and Technology
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• 제63권1호
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• pp.36-45
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• 2021

Presently, there is an increased demand for livestock products all over the world which has led to more devotion on improving livestock population. Although goats have been bred for a long time in Korea, but there is not much research conducted on traditional Korean black goat (Capra hircus coreanae) compared to other livestock populations. Mutton consumption has been dramatically changing from medicinal use to edible meat and this trend directs the black goat populations declining and also mutton import quantities are increasing consistently. The present study introduced a new estrus synchronizing technique with subsequent artificial insemination (AI) for Korean black goats to enable crossbreeding with non-native breeds for the small or subsistent farmers. Our data highlighted that, the percentage of motile sperm from the electro-ejaculated samples declined significantly after freezing and melting. In addition, the sperm motility significantly declined with regard to sperm incubation period (0, 5, 60, and 120 min at 37℃) and was negatively correlated (64.2 ± 7.9%, 63.3 ± 5.8%, 49.9 ± 6.3%, and 35.9 ± 7.6%, respectively) in frozen-thawed sperm samples. Moreover, the E2 levels were unchanged even 24 h after controlled internal drug releas (CIDR) withdrawal. But, 48 h and 72 h after CIDR removal, E2 levels increased significantly. These data helps us to consider the two time points for AI; CIDR removal after 24 h, at which E2 decreases, and after 48 h, as the time at which progesterone increases. Additionally, the AI after 48 h of CIDR removal group exhibited significantly higher pregnancy and parturition rates (42.9%) compared to AI after 24 h after CIDR removal 28.6% group. In conclusion, these studies will propose an optimal estrus synchronisation process with subsequent timing of AI and also will promote the Korean black goat breeding industry.

• 동물자원연구
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• 제29권4호
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• pp.158-165
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• 2018

본 연구는 Canada에서 수입한 동결정액으로 인공수정된 모돈의 번식성적을 조사하여 동결정액을 이용한 인공수정(동결정액 인공수정)의 효율을 개선할 수 있는 통찰력을 얻기 위해 수행되었다. 이를 위해 A(GGP) 농장에서 2016년 5월과 9월 사이 총 626회의 동결정액 인공수정과 B(GGP) 농장에서 2015년부터 2017년 기간 동안 수행한 총 2,429 동결정액 인공수정의 결과 기록을 분석하였다. A종 돈장에서 동결정액을 썼을 때 총산자수 및 실산자수는 9월 보다 5월에 높았다(p<0.05). B종돈장의 분만율, 총산자수 및 실산자수는 조사연도 간에 차이가 없었다. A종돈장 결과와 B종돈장 결과를 통합하여 분석했을 때, 분만율은 A종돈장이 높았으나(p<0.01) 총산자수와 실산자수는 두 종돈장간에 차이가 없었고, 동결정액 인공수정을 했을 때가 액상정액 인공수정을 했을 때보다 낮았다(동결정액:액상정액 인공수정에 대한 총산자수와 실산자수는 각각 $10.9{\pm}0.3:13.4{\pm}0.1$두 및 $10.0{\pm}0.3:12.0{\pm}0.1$두 이었음; p<0.01). 결론적으로, 이상의 결과는 동결정액 인공수정이 액상정액 인공수정보다 번식 효율이 낮긴 하지만 연중 적절한 시기에 동결정액 인공수정을 실시하면 번식 효율이 증가될 수도 있음을 시사한다.

• Reproductive and Developmental Biology
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• 제28권3호
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• pp.167-172
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• 2004

Flow cytometry를 이용하여 개 정자의 생존율 평가를 수행하고자 2-4세의 수캐 5두가 이용되었고, 분석을 위해 PI염색을 실시하였다. Flow cytometry를 이용한 개 신선 정액의 생존율 평가는 생존 정자와 죽은 정자의 비율을 1:0, 1:1, 1:3으로 조성하여 이를 flow cytometry로 평가하고 광학현미경검사, CFDA/PI 염색검사, HOS test에 의한 생존율과 비교하여 flow cytometry와의 상관관계를 알아보았다. 또한 개 정액을 동결하여 응해 후의 개 정자의 생존율 평가에도 동일한 방법으로 상관관계를 조사하였다. 신선 정액에서 생존 정자와 죽은 정자의 비율이 1:0, 1:1, 1:3 모든 경우에서 flow cytometry를 이용한 생존율은 HOS test에 의한 생존율과 높은 상관관계를 나타내었다 (p<0.01). 신선 정액에서 생존 정자와 죽은 정자의 비율이 1:0과 1:3일 때 광학현미경적 검사에 의한 생존율은 flow cytometry 분석에 의한 생존율과 유의 적인 상관관계를 나타내었으나 (p<0.05), 1:1 비율의 경우 상관관계를 보이지 않았다. 신선 정액에 생존 정자와 죽은 정자의 비율이 1:0과 1:1일 때 CFDA/PI 염색 검사에 의한 생존율은 flow cytometry분석에 의한 생존율과 높은 상관관계를 보였으며(p<0.01), 1:3 비율에서는 유의적인 상관관계를 보였다 (p<0.05). 동결 및 응해 후의 개 정자의 생존율 평가에서 HOS test 결과는 flow cytometry분석에 의한 생존율과 높은 상관관계를 보였으며 (p<0.01), 광학현미경적 검사를 통한 생존율은 유의적인 상관관계를 보였으나 (p<0.05), CFDA/PI 염색 검사결과는 상관관계를 보이지 않았다. 이상의 결과 flow cytometry는 신선정액 및 동결 융해 후 개 정자에 대한 생존율 검사에 정확한 평가 방법 인 것으로 판단되었다.

• 한국가금학회지
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• 제41권1호
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• pp.21-27
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• 2014

본 연구는 닭 정액의 동결 보존을 위하여 비 글리세롤성 동결 보호제 중 MA 농도가 정자의 생존율과 융해된 동결 정자를 인공수정을 실시하여 생산된 수정란의 수정율, 부화율을 조사하고자 실시하였다. 동결 보조제로써 MA의 효율성은 7%, 9% 및 11% 범위에서 동결을 실시하였을 때, 정자의 생존율은 $35.16{\pm}6.12%$, $67.83{\pm}15.3%$ 및 $66.2{\pm}16.3%$로 관찰되었으며, 융해된 정자를 인공수정을 실시하여 생산된 수정란의 수정율은 21.5%, 34.7% 및 25%로 관찰되었으며, 수정된 수정란의 부화율은 100%, 89.5% 및 87.5%로 관찰되었다. 대조군으로써 신선 정액은 수정율이 96.0%로 관찰되었고, 부화율은 92.2%로 관찰되었다. 9% MA를 이용한 간이 동결법으로 생산된 동결 정자를 이용하여 3주간 수정란을 검사하였을 때, 수정율은 비록 35.3%로 관찰되었으나, 부화율은 90.3%로 관찰되었다. 이러한 결과에 따르면, 9% 농도로 MA 동결 보호제를 이용할 경우, 동결 및 융해된 정자를 이용하여 생산된 수정란에서 수정율을 감소시킬 수 있음을 보여주고 있으며, 부화율에는 영향을 주지 않음을 추정할 수 있다. 그러므로 수정율과 부화율에 나쁜 영향을 미치지 않고 가금 유전자원의 보존에 중요한 요인이 될 수 있는 적절한 농도의 MA 동결 보호제 범위는 7~9% 농도로 추정된다.

• 한국수정란이식학회지
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• 제18권2호
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• pp.143-149
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• 2003

한우의 유전자원을 보존하기 위하여 정액동결 시험을 수행하였으며 고급육계통은 육질에 대한 육종가 상위 10% 이내의 자손에서 선발하였고 다 유계통은 어미의 이유시 체중에 대한 육종가 상위 10%이내의 자손에서 선발하였으며 2개 계통에서 종모우 총 13두를 선발하여 공시하였다. 정액채취는 인공질법으로 실시하였고 의빈대에 수소를 계류하고 채정대상우를 승가시켰으며, 3회 가승가후에 채정하였다. 채정후 10분 이내에 실험실로 옮겨와 검사항목을 조사하고 37$^{\circ}C$에서 1차 희석을 하고 5$^{\circ}C$까지 하강한 후 2차 희석을 하였으며, 액체 질소 5cm 위에서 5분간 평형후 침지하여 동결하였다. 이들 종모우 정액의 일반적 성상에 있어서 채정된 정액량은 1차와 2차를 합하였을 때 평균 채정량이 5.7 ml였고 정자농도는 975${\times}$$10^{6}$개였으며 정액의 색상은 유백색이었고 pH는 6.8이었다. 채취시 생존성은 90.2% 그리고 동결 융해후에는 65.7 %가 생존하였으며 총 6,870스트로우의 동결 정액을 생산하여 보존하였다.