• Clinical and Experimental Reproductive Medicine
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• 제42권2호
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• pp.33-44
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• 2015

The generation of artificial gametes is a real challenge for the scientific community today. In vitro development of human eggs and sperm will pave the way for the understanding of the complex process of human gametogenesis and will provide with human gametes for the study of infertility and the onset of some inherited disorders. However, the great promise of artificial gametes resides in their future application on reproductive treatments for all these people wishing to have genetically related children and for which gamete donation is now their unique option of parenthood. This is the case of infertile patients devoid of suitable gametes, same sex couples, singles and those fertile couples in a high risk of transmitting serious diseases to their progeny. In the search of the best method to obtain artificial gametes, many researchers have successfully obtained human germ cell-like cells from stem cells at different stages of differentiation. In the near future, this field will evolve to new methods providing not only viable but also functional and safe artificial germ cells. These artificial sperm and eggs should be able to recapitulate all the genetic and epigenetic processes needed for the correct gametogenesis, fertilization and embryogenesis leading to the birth of a healthy and fertile newborn.

• 한국수정란이식학회지
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• 제25권2호
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• pp.97-101
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• 2010

Intracytoplasmic sperm injection (ICSI) is one of the artificial fertilization methods when only a few sperm are available for insemination, and an important tool for the preservation of genetic materials of endangered animal species, especially the male is infertile. Different from other species such as mice and pigs, the conventional ICSI method which uses spiked pipette for injection (Spike-ICSI) is exhibited low success rates in cattle because the bovinesperm head membrane is hard to break during injection procedure. We chose piezo-assisted ICSI (Piezo-ICSI) for the improvement of the injection procedure including sperm head membrane rupture and efficient puncture of the plasma membrane of the oocytes. In this experiment, we compared the efficacy of the bovine ICSI embryo production between the Piezo-ICSI and Spike-ICSI. The second polar body extrusion, pronuclear formation, cleavage and blastocyst formation were evaluated after implementation of two different ICSI techniques. The Piezo-ICSI tended to show comparably higher rates of the second polar body extrusion (41.7%), the pronuclei formation (42.9%) and the two-cell cleavage (41.4%) than Spike-ICSI does (33.3%, 28.6% and 23.5%, respectively) although there is no statistic significance between two groups. In addition, the blastocysts were only obtained from the Piezo-ICSI group (10.3%). Our finding shows that the Piezo-ICSI may be used as an artificial fertilization method in cattle when in vitro fertilization is not applicable.

• Asian-Australasian Journal of Animal Sciences
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• 제27권10호
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• pp.1417-1425
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• 2014

The selection of morphologically normal spermatozoa is critical to obtain high breeding performances in boar breeding farms and artificial insemination (AI) centers. Parameters for the selection of semen mainly include total sperm motility, concentration, and morphology. However, these primary parameters are often not reliable for discriminating between normal and abnormal, non-fertilizable spermatozoa. The present study was designed to compare the motion characteristics, fertilization ability using in vitro fertilization (IVF), and acrosome formation of the semen from boars having low (boar number 2012) and normal (boar number 2004 and 2023) breeding performances. The ultimate goal was to identify additional simple and easy criteria for the selection of normal sperm. There was no significant difference between boar 2004 and boar 2023 sperm total motility in computer assisted sperm analysis. However, boar number 2012 semen presented a significantly reduced population of rapid moving spermatozoa and an increased population of slow moving spermatozoa compared to boar numbers 2004 and 2023. Analysis of detailed motion characteristics revealed that sperm from boar number 2012 had significantly reduced motility in progressiveness, average path velocity, straight-line velocity (VSL), curvilinear velocity (VCL), straightness, and linearity. The assessment of the fertilizing ability by IVF also showed that sperm from boar number 2012 showed a fertility rate of 3.4%, whereas sperm from boar number 2023 had a fertility rate of 75.45%. Interestingly, most of the sperm nuclei were found on the peripheral area of the oocytes, suggesting that the sperm from boar number 2012 lacked penetration ability into the oocyte zonapellucida. The acrosome formation analysis using Pisum sativum agglutinin staining demonstrated that the sperm from boar number 2012 had a defect in acrosome formation. Consequently, primary parameters for selecting semen before AI such as motility are not sufficient to select normal and fertilizable spermatozoa. In conclusion, the present study suggests that the acrosome staining and detailed motion characteristics such as progressiveness, VCL, and VSL should be included in determining semen quality together with primary parameters for successful AI and high breeding performance in the swine industry.

• 대한한방부인과학회지
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• 제24권4호
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• pp.194-204
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• 2011

Purpose: Many infertile women are receiving in vitro fertilization-embryo transfer (IVF-ET). But side effects occur after IVF-ET and pregnancy rate is still low. So this study is to report the effect of herbal medicine on a woman receiving IVF-ET and herbal medicine's low risk on a pregnant woman. Methods: The patient was underwent artificial insemination five times and IVF-ET three times. In this study, the patient steadily took a herbal medicine before IVF-ET to birth. Results: The side effects of IVF-ET and the miscarriage symptoms ware decreased. And by taking a herbal medicine during IVF-ET, the patient was pregnant and gave birth when she was 33 weeks pregnant by cesarian section. Conclusions: This case study shows that herbal medicine is effective for reducing side effects of IVF-ET and increasing pregnancy rate. And in this case the patient gave birth to healthy babies, althogh steadily taking herbal medicine during pregnancy.

• 한국수정란이식학회지
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• 제27권1호
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• pp.29-35
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• 2012

The purpose of this study was to examine the effect of growing stages of the Korean Native Striped Bull (KNSB) on the freezability and fertility of frozen-thawed semen. First, we investigated the total motility (TM) and progressive motility (PM) according to the diluent used for semen freezing. Second, we examined the effect of the age of KNSB on semen volume, TM and PM of fresh and frozen-thawed semen. Third, we examined the effect of frozen semen from the different age of KNSB on the $in-vitro$ fertilization rate, and the artificial insemination pregnancy rate. The diluents used in this experiment were Triladyl$^{(R)}$ and Tris-egg yolk extender (EYE). Semen was collected from 5 KNSB in the growing stage (15 months) and 5 adult KNSB (36 months). When Triladyl or Tris-EYE extender was used for semen freezing, there was no difference of the mean TM and the mean PM. However, the mean TM was significantly higher in Bull No. 1885 than Bull No. 4283 ($p$ <0.05). The mean volume of semen collected from the 15-month-old bulls (2.3 ml) was significantly lower ($p$ <0.05) than that from the 36-month-old bulls (5.0 ml). The mean semen concentration was similar for the 15-month-old ($2.1{\times}10^9$ spermatozoa/ml) and 36-month-old ($1.8{\times}10^9$ spermatozoa/ml) bulls. For the 15-month-old and 36-month-old bulls, the mean TM of fresh semen were 93.7% and 88.3%, respectively, and the mean PM were 97.0% and 88.3%, respectively; the 15-month-old bulls showed a particularly high PM ($p$ <0.05). For the 15-month-old and 36-month-old bulls, the mean TM (56.0% and 58.0%, respectively) and the mean PM (64.0% and 70.7%, respectively) of frozen-thawed semen did not differ. The development rates of embryos after $in-vitro$ fertilization and the pregnancy rate after artificial insemination using frozen-thawed semen did not differ according to the bull's age. In summary, semen volume differed according to the bull's age, but semen concentration and survival rate, the $in-vitro$ fertilization rate, and the pregnancy rate did not differ according to the stripe bull's age. Accordingly, semen from bulls in the growing stage can be collected and frozen for the preservation and multiplication of rare livestock.

• 한국자원식물학회지
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• 제31권4호
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• pp.372-377
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• 2018

본 연구는 천마의 종자생산을 위한 수확시기별 처리 방법에 따른 자마 생산 효과를 구명하고자 수행하였다. 봄수확 천마는 괴경의 무게를 60-99 g, 100-139 g, 140-179 g, 180 g 이상으로 처리하였고, 가을수확 천마는 $5{\pm}1^{\circ}C$의 저온을 2, 4, 6, 8, 10주 처리하였다. 그 결과, 봄수확 천마는 괴경의 무게가 100 g 이상일 경우, 인공수정률이 90.9%-94.8%, 원괴체 형성률이 3.1%-5.4%, 자마 형성률이 10.1%-15.3%로 나타났다. 가을수확 천마는 4주 이상의 저온처리 시 인공수정률이 70.4%-87.6%, 원괴체 형성률이 2.2%-2.6%, 자마 형성률이 8.7%-9.5%로 나타났다. 따라서, 종자 및 자마 생산을 위해서는 봄수확 천마는 100 g 이상의 괴경을 사용해야 하고, 가을수확 천마는 저온에서 4주 이상의 휴면타파가 필요했다. 가을수확 천마를 이용할 경우 봄수확 천마에 비해 인공수정률, 원괴체 및 자마 형성률이 낮았지만, 자연 발아 상태보다 월등히 높았다. 따라서 일정한 괴경의 무게와 저온처리 휴면타파를 이용하면 천마의 종자와 자마가 확보됨으로써 천마의 연중생산이 가능할 것으로 판단되었다.

• 한국양식학회지
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• 제10권2호
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• pp.113-121
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• 1997

강원연안에 서식하는 코끼리조개의 인공종묘생산 기술개발을 목적으로 산란유발 방법 및 수정과 부화를 위한 수온과 염분농도의 최적조건 등을 연구한 결과를 요약하면 다음과 같다. 코끼리조개의 산란유발은 간출자극과 자외선조사해수자극에서는 전혀 반응이 없었다. 수온자극에서는 5월에 15.0-25.0%의 반응을 보였고, 6월에는 10.0%의 반응을 보였다. 그러나 생식소를 절개한 것은 15.0-45.0%로 가장 높은 반응을 보였다. NH 하(4) OH 용액을 해수에 첨가하는 자극에서는 8/1000N-10/1000N에서 15.0%의 반응율을, 그리고 NH 하(4) OH 용액을 생식소에 주사하는 자극은 5/100N-7/100N에서 5.0-10.0%의 반응율을 보였다. 코끼리조개의 수온에 따른 수정율은 수온 11-17$^{\circ}C$범위에서 74.2-89.2%였고, 부화율은 수온 8-$14^{\circ}C$에서 84.3-90.5%로 가장 높았다. 염분 농도에 의한 수정율은 염분 25-$35tetperthousand$범위에서 72.5-88.5%, 부화율은 염분 30-$35tetperthousand$에서 82.7-86.9%였다. 수정과 부화를 위한 수온과 염분농도의 최적조건은 수온 11-$14^{\circ}C$, 염분농도 30-$35tetperthousand$로 나타났다.

• 한국수산과학회지
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• 제54권1호
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• pp.31-37
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• 2021

The mass production of fertilized eggs of the convict grouper Hyporthodus septemfasciatus was studied from 2013 to 2020 for industrial aquaculture. The experiment was divided into two groups. Group 1 broodstock was raised from wild-caught fry and used from 2013 to 2020. Group 2 broodstock was raised from artificially propagated fry and used from 2019 to 2020. Males used to collect sperm for artificial insemination weighed more than 7 kg. The effects of various hormones on artificial ovulation were investigated from 2013 onward. Among these, luteinizing hormone-releasing hormone analogue (LHRHa) at 100 ㎍/kg body weight showed the most effective results and was used for artificial egg collection from 2014 onward. In Group 1, the average total egg production per year, average egg production per individual, fertilization rate, and hatching rate were 26,143 mL, 609.7 mL, 93.3%, and 91.8%, respectively, and in Group 2, were 2,750 mL, 316.5 mL, 92.1%, and 90.4%, respectively. Based on these results, we showed that a large number of fertilized eggs for artificial seeding could be produced consistently. Moreover, the mass production of fertilized eggs in Group 2 establishes a foundation for the complete aquaculture cycle of H. septemfasciatus.

• 한국수산과학회지
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• 제8권3호
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• pp.185-195
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• 1975

1. 본 논문은 1969연부터 1974연에 이르기까지 인천 송도, 안면도, 부안 근해에서 채집한 대합 M. lusoria와 가무락 C. sinensis의 산란 유발, 초기 발생 및 유생 성장에 관한 연구를 한 것이다. 2. 대합은 4/100와 5/100N의 $NH_4OH$ 해수용액을 난소에 직벚 주사하여 $25.0\~33.3\%$ 산란유발이 가능했고, 가무락에서는 1/100, 1/180, 3/100 N의 $NH_4OH$의 해수 용액으로 $12.5\~19.0\%$의 산란 유발을 할 수 있었다. 그러나 가무락의 경우는 반복온도 자극에 의해서 $40.0\%$까지 방란 및 방정 유발을 가능하게 했다. 3. 대합과 가무락에 있어서의 각각 $36.9\%$와 $39.3\%$였다. 4. 1/1000N $NH_4OH$, 해수용액에 의한 대합과 가무락의 초기 발생과정에서는 수정란 50분에 극체가 출현하고 5시간 40분이 경과되면 담륜자 유생으로 되고 2시간이 지나면 D형 피면자 유생이 됨을 관찰하였다. 5. 두 종의 유생을 사육한 결과 대합은 수정 후 1일이 지나면 평균 각장이 $10.9.5{\pm}0.7\mu$인 초기 D형기에, 11일 경과하면 $14.6{\pm}1.3\mu$ 인 각정기에, 20일이면 $208.0{\pm}0.0\mu$인 변태기에 달하였고, 가무락은 1일이 지나면 평균 각장이 $110{\mu}5{\pm}0.6\mu$인 초기 D형기에, 10일 이면 $141.8{\pm}1.7$인 각정기에, 20일이면 $235.0{\pm}0.0\mu$의 변태기에 달함을 관찰하였다. 6. 초기 D형으로부터 변태기까지의 각장(L)과 각고(H)의 상대성장 관계는 대합이 H=0.77L+6.82 이고, 가무락은 H=0.75L+8.50의 식으로 나타낼 수 있다.

• Asian-Australasian Journal of Animal Sciences
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• 제11권2호
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• pp.192-195
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• 1998

Biomass conversion efficiencies (B.C.E) of six fish species viz, Catla catla, Labeo, rohita, Cirrhina mrigala, Hypophthalmicthys molitrix, Ctenopharyngodon idella and Cyprinus carpio cultured under artificial feed (T1), broiler manure (T2), buffalo manure (T3), N:P:K (25:25:0) (T4) and control pond (T5) have been determined for the period of one year. The overall biomass conversion efficiencies under the influence of T2, T3, and T4 were statistically similar. However, the best (0.40) efficiency was determined under feed supplement-ation (T1).