• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.239-246
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• 1997

The objective of this study was to evaluate the variation of fertilizing ability following the morphologically normal sperm ratio in porcine IVF using epididymal sperm The results obtained in this experiment were summarized as follows: 1. When the penetration rate (PR), polysper my rate (PSR), pronuclei formation (2PNF) and mean number of sperm (MNS) per oocyte were evaluated according to the percentage of morphologically normal epididyrnal sperm at insemination($\leq$lO%, 10~30% and $\geq$50%). the PR and PSR of $\leq$50% group (82.4, 87.4%) were significantly higher than those of other two groups ($\leq$lO%; 29.7%, 22.6% and 10~30%; 20.3, 37.0%) (p<0.01). Also, the 2PNF per examined oocytes was significantly high in $\geq$ 50% group (p<0.01). 2. When the $\geq$50% group in epididymal sperm was adjusted to 100% (5x1$^5$ cells/ml) , the PSR and 2PNF were not different between epididymal sperm (86.7, 35.1%) and frozen-thawed ejaculated sperm (86.0. 39.4%) although the PR in epididymal sperm (79.7%) was significantly lower than that in frozen-thawed ejaculated sperm (95.5%)(p<0.01). 3. Also. when the PR, PSR, 2PNF and MNS of epididymal sperm were evaluated according to the oocyte: sperm ratio (1:6000, 1:6650. 1:7700 and 1: 10000) at insemination. the PR, PSR and MNS were increased as the oocyte:sperm ratio increases. However, this result indicated that the 2PNF was high in the oocyte:sperm ratio (1:6000 and 1:6650). Therefore. these results suggested that when the percentage of morphologically normal epididymal sperm was more than 50. the fertilizing a ability was very similar to that of frozen-thawed ejaculated sperm and that the detailed evalu¬a ation of morphological normality in porcine IVF using epididymal sperm should be prerequisite to obtain the more effective fertilizing ability.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.19-33
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• 2000

Eighty dairy farms (38 farms responded) in Palmerston North area of New Zealand were questioned by mail questionnaire on: 1) general characteristics, 2) milk yield and feed supplementary, 3) reproductive efficiencies (12 questions) and 4) reproductive disorders (12 questions) from February to July, 1998. Among those 4 items, the reproductive efficiencies (3) and disorders (4), various diseases and culling rates were surveyed and analyzed for Korean dairy farmers (especially in Cheju island) and compared with New Zealand. The results are as follows: 1. Fifteen farms in 38 dairy farms relied entirely on artificial insemination, the rest of 23 dairy farms (60.5%) raised 5∼6 bulls to increase conception rate. The dairy farmers in Palmerston North used artificial insemination from Oct 4th to Dec 10th for 42.8 days, and then used bulls from that point to coming Jan 10th for 41.4 days. The submission rate within 3, 6 and 10 weeks following the initiation of AI season was 84.7, 93.9 and 97.9% respectively. 2. The average age of heifers at the first estrus, pregnancy and calving was 11.0, 18.0 and 24.7 months respectively, and an average 1.4 estrus cycles were required for conception. The intervals of estrus recurrence and the following conception after calving were 38 and 68 days respectively. 3. Among inseminated cows, calving, abortion and empty cow was 90.9, 1.6 and 7.4% respectively. Calving rate decreased according to increasing farm size, while the number of empty cows decreased. 4. Stillbirth, retained placenta and delivery abnormalities were 5.3, 3.7 and 5.5% respectively, not different depend on herd size. 5. The incidence of milk fever, grass tetany, and ketosis was 3.6, 3.0 and 1.0%, respectively. The delivery abnormality and mastitis treated with medicine were 3.1 and 6.7%, but decreased according to farm size. Lameness was 8.6% on average, but over 10% in farms which has more than 400 milking cows. 6. Among the culled cows (15.5% of the total), those culled due to an old age, lameness and other diseases were 2.9, 1.8 and 4.3% respectively and those culled due to low milk production, reproductive abnormality reduced with farm size. 7. Compared with the data collected in Korea, the reproductive efficiency was better, and lameness, metabolic problem and culling rate were higher in New Zealand

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.97-108
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• 2000

The objectives of this study were performed to increase the efficiency of the culture conditions of embryos produced in vitro, and to assess the developmental potential after transfer of those embryos into recipients. The mean number of folliclular oocytes recovered from an ovary was 10.7. The rates of maturation and fertilization in Grade I oocytes were significantly (P＜0.05) higher than Graden and III. Developmental rate into blastocyst in the culture group of TCM-199 with BOEC were significantly higher (P＜0.05) than the groups of TCM-199 and conditioned medium (24.7% vs. 12.4% and 18.2%). The survivability of post-thawed blastocysts equilibrated for 3 min in EFS solution was significantly (P＜0.05) lower than l0 for 1 and 2 min (32.1% vs. 82.9% and 73.3%). Significantly higher (P＜0.05) survival rate in blastocysts was seen after freezing than in morulae stage embryos. Out of all 105 recipients, 49 (46.7%) were confirmed in pregnant. On pregnancy of cattle, 48 calves were born from 40 recipients. The ratio of twin and single calves was 30.5% (32/40 and 7.6% (8/40), respectively. However, the others composed of abnormal, as judging as 6 (12.2%) for abortion and 3 (6.1%) for stillbirth during the pregnant period.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.6
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• pp.702-708
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• 2002

Although Umboniunm thomasi is one of marine mollusc (Archaeogastropoda: Trochidae) inhabiting the sands in the intertidal zone of the west coast of Korea, aspects of its reproductive biology are still not too well known. Reproductive cycle, gametogenesis, and first sexual maturity of U. thomasi collected at the west coast of Buan-gun, Jeollabuk-do, Korea were investigated monthly from January to December 1999. U. thomasi was dioecious, and an oviparous. The gonad was placed in the rear of the flesh part in the spiral shell. The external colors of the ripe ovary and testis appeared to be green and milk-white or yellowish white, respectively. Meat weigh rate peaked in July ($37.5\%$). And then the value sharply decreased in September ($28.3\%$), thereafter, gradually increased in November ($31.7\%$). Fully ripe oocytes were approximately 100$\~$110 $/mu$m in diameter, and their cytoplasm contained a great number of yolk Branules. Based on the monthly changes of the Bonadal development, gametogenesis, and meat weight rate, the reproductive cycle of U. thomasi could be devided into five successive stages: early active (November to April), late active (February to May), ripe (April to August), spawning (July to October), and recovery (September to February). Gonadal development and spawning were closely related to the seawater temperature, the main spawning occurred in September when the temperature reached above 24.2$^{\circ}C$. Individuals of 4.4 mm and less in shell height could not take part in reproduction in both sexes. Percentages of first sexual maturity of female and male shells ranging from 5.5 to 6.4 mm were $55.0\%$ and $61.9\%$, respectively, and $100\%$ of those over 7.5 mm in shell heights in both sexes participated in the reproduction.

• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.141-148
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• 1999

This study was to test whether the viability of bovine hatched blastocysts (HBs) can be maintained after vitrification and thawing. The HBs were produced in vitro at Day 9 and Day 10 after IVF, and they were classified to small (S-HBs; ø$\leq$300 ${\mu}{\textrm}{m}$) and large(L-HBs; ø＞300 ${\mu}{\textrm}{m}$) on the basis of embryo diameter using eyepiece micrometer. As freezing solution, we used EFS35 which consisted of 35% ethylene glycol (EG), 18% ficoll, 0.3 M sucrose and 10% FBS added in mDPBS. Vitrification was taken by two-step method, the HBs were equilibrated in 10% EG for 5 minutes and then shortly exposed in EFS35 and plunged into L$N_2$for 30~45 sec. After thawing, the survival rates were assessed by the re-expansion of the blastocoel during 2 h and 16 h of culture. The results obtained in these experiments were summarized as follows; 1) When the blastocysts(40.8%) recovered at Day 8 after IVF were further cultured for 24 h(Day 9 after IVF) and 48 h(Day 10 after IVF), the rates of HBs were 20.5% and 6.7%, respectively. Also, the total cell number of HBs on Day 9 was significantly higher than that of HBs on Day 10 (p＜0.01). 2) When the effects of freezing solution to the survival of Day 9 L-HBs were examined, the rate of vitrified group (75.7%) was significantly lower than 100% of control and exposed group(p＜0.05). 3) When the survival rates of vitrified HBs according to size and developmental age were examined, the data of L-HBs (75.5%) and S-HBs(63.6%) on Day 9 were slightly higher than those of L-HBs(64.3%) and S-HBs(60.7%) on Day 10. 4) Also, when the in vitro survival of Day 9 HBs was evaluated under different culture condition after thawing, the result in culture medium only (79.3%) was significantly higher than 43.2% in co-culture group (p＜0.05). These results demonstrated that bovine HBs can be successfully cryopreserved by two-step vitrification method using EFS35.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.407-417
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• 2000

The present study was to examine effects of various electrical stimulus treatments used for electro-fusion on the preimplantation development of bovine nuclear transfer (NT) embryos with fetal fibroblast cells. Fetal fibroblast cells were isolated from one fetus at day 45 of gestation in Holstein cow, and passaged 3 to 4 times before being transferred into enucleated oocytes. Single fibroblast cells were individually placed into the perivitelline space of enucleated oocytes by using a micromanipulator. At first, the fusion and developmental rates of reconstructed oocytes were compared between different electric stimulation conditions. When fusion of the reconstructed oocyte was induced by different electric pulse periods (15, 30 and 45 $\mu$sec) at a DC pulse of 1.8 kV/cm, 15 (45.5%, 120/264) or 30 $\mu$ sec group (43.9%, 106/241) showed a higher fusion rate than 45 $\mu$sec group (23.2%, 58/250, P＜0.05). However, no difference was detected in the development rate of the fused oocytes to blastocysts between groups. Next experiment was to examine the effects of different electrical field strengths (1.5, 1.8 and 2.1 kV/cm) for 15 $\mu$sec at electrofusion on in vitro development of the NT embryos. As results, there was no difference in the fusion and developmental rates of the NT embryos between electrical strength (P＞0.05). Finally, developmental competence of bovine NT embryos with somatic cells was compared with IVF-derived embryos. Of enucleated oocytes fused with fibroblast cells, 27.4% (75/274) developed to the blastocyst stage, which is similar to that (24.5%, 58/237) of IVF-derived embryos. However, mean nuclei number of NT blastocysts was smaller than that of IVF-derived blastocysts. Thus, we have established an optimal condition (1.8 kV/cm, 15 $\mu$sec) for electric fusion of bovine NT oocytes with somatic cells. The present study indicates that bovine reconstructed embryos with somatic cells normally develop to blastocyst stage in vitro, although having smaller nuclei numbers of blastocysts as compared to IVF-derived embryos.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.49-57
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• 2000

This study was conducted to investigate the developmental potential of cloned embryos derived from bovine fetal fibroblast cells, and the effect of quiescent treatment, passage number and origin of donor cells on in vitro development of cloned embryos. Fetal skin and liver-derived fibroblast cells were transferred to enucleated oocytes after serum starvation or nontreatment (cycling). After electrofusion. reconstituted embryos were activated with $Ca^{＋＋}$-ionophore and cycloheximide, and cocultured for 7~9 days with BRL cells. Some blastocysts were transferred to recipient cows 7~8 days post estrus. The development rate to the blastocyst stage of serum starved cell-derived embryos was higher (25.3%) than that of actively dividing cells-derived embryos (15.9%), The rates of blastocyst formation were 23.1~25.0% after transfer of cell passaged 4 to 6 times, and 23.8 and 25.2% after transfer of fetal skin and liver cells, respectively. After embryo transfer, 34.4% and 15.6% of recipient cows were pregnant on Day 60 and 120, respectively, and one male calf was produced from skin-derived vitrified blastocyst. The result of this study showed that the development of cloned embryos. was enhanced by quiescent treatment, but did not different among the cells passaged 4 to 6 times, and between skin and liver cells. This result also confirms that offspring can be obtained from the vitrified clone embryo derived from fetal skin cell.

• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.327-337
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• 2001

This study investigated the effects of superoxide dismutase (SOD) on lipid peroxidation and fertilizing ability in vitro of boar spermatozoa frozen-thawed. The percentages of motile sperm were highest when SOD of 10 units/$m\ell$ was added to washing medium for spermatozoa. However, the rates of motile sperm were not significantly different in different concentrations of SOD. On the other hand, the motile rates of sperm washed with SOD were lower in sperm inculbated for 120 min than 30 min regardless of the different concentrations of SOD. The percentage of spermatozoa that reached acrosome reaction were increased with incubation periods prolonged. No significant differences, however, were observed in acrosome reaction rates between sperm incubated with and without SOD supplementation for 0, 60 and 120 min. When oocyies inseminated with different concentrations of SOD, the penetration rates were significantly (P＜0.05) higher in medium with 1 unit/$m\ell$ than 0, 10 and 100 units/$m\ell$ of SOD. However, the proportions of polyspermit oocytes were significantly (P＜0.05) lower in medium with 10 and 100 units/$m\ell$ than 0 unit/$m\ell$ of SOD. In another experiment, the sperm suspension were also treated with different concentrations of SOD and were assayed far sulfhydryl(-SH) group content. In the groups treated with 100 units/$m\ell$ of SOD, sperm-SH group were higher than another groups. However, sperm-SH group content were not siginificantly different in spermatozoa treated with different concentrations of SOD. Under the same conditions, the lipid peroxidation of sperm was evaluated on the basis of malondialdehyde production. The addition of SOD to sperm suspension decreased the formation or malondialdehyde. However, there were not significantly different in sperm treated with different concentrations of SOD. The activity of sperm binding to zona pellucida was also evaluated through binding to salt-stored porcine oocytes. The sperm binding to zona pellucida were gradually increased with SOD concentrations added. The number of spermatozoa binded to zona pellucida were significantly (P＜0.05) higher in medium with 100 units/$m\ell$ than 0 units/$m\ell$ of SOD. These findings suggested that SOD cause an enhancement penetrarion ability and sperm zona binding in boar spermatozoa frozen-thawed.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.1
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• pp.124-136
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• 2002

As an official journal of the Asian-Australasian Association of Animal Production Societies (AAAP), the Asian-Australasian Journal of Animal Sciences (AJAS) was born in February 1987 and the first issue (Volume 1, Number 1) was published in March 1988 under the Editorship of Professor In K. Han (Korea). By the end of 2001, a total of 84 issues in 14 volumes and 1,761 papers in 11,462 pages had been published. In addition to these 14 volumes, a special issue entitled "Recent Advances in Animal Nutrition" (April, 2000) and 3 supplements entitled "Proceedings of the 9th AAAP Animal Science Congress" (July, 2000) were also published. Publication frequency has steadily increased from 4 issues in 1988, to 6 issues in 1997 and to 12 issues in 2000. The total number of pages per volume and the number of original or review papers published also increased. Some significant milestones in the history of the AJAS include that (1) it became a Science Citation Index (SCI) journal in 1997, (2) the impact factor of the journal improved from 0.257 in 1999 to 0.446 in 2000, (3) it became a monthly journal (12 issues per volume) in 2000, (4) it adopted an English editing system in 1999, and (5) it has been covered in "Current Contents/Agriculture, Biology and Environmental Science since 2000. The AJAS is subscribed by 842 individuals or institutions. Annual subscription fees of US$ 50 (Category B) or US$ 70 (Category A) for individuals and US$ 70 (Category B) or US$ 120 (Category A) for institutions are much less than the actual production costs of US$ 130. A list of the 1,761 papers published in AJAS, listed according to subject area, may be found in the AJAS homepage (http://www.ajas.snu.ac.kr) and a very well prepared "Editorial Policy with Guide for Authors" is available in the Appendix of this paper. With regard to the submission status of manuscripts from AAAP member countries, India (235), Korea (235) and Japan (198) have submitted the most manuscripts. On the other hand, Mongolia, Nepal, and Papua New Guinea have never submitted any articles. The average time required from submission of a manuscript to printing in the AJAS has been reduced from 11 months in 1997-2000 to 7.8 months in 2001. The average rejection rate of manuscripts was 35.3%, a percentage slightly higher than most leading animal science journals. The total number of scientific papers published in the AJAS by AAAP member countries during a 14-year period (1988-2001) was 1,333 papers (75.7%) and that by non- AAAP member countries was 428 papers (24.3%). Japanese animal scientists have published the largest number of papers (397), followed by Korea (275), India (160), Bangladesh (111), Pakistan (85), Australia (71), Malaysia (59), China (53), Thailand (53), and Indonesia (34). It is regrettable that the Philippines (15), Vietnam (10), New Zealand (8), Nepal (2), Mongolia (0) and Papua New Guinea (0) have not actively participated in publishing papers in the AJAS. It is also interesting to note that the top 5 countries (Bangladesh, India, Japan, Korea and Pakistan) have published 1,028 papers in total indicating 77% of the total papers being published by AAAP animal scientists from Vol. 1 to 14 of the AJAS. The largest number of papers were published in the ruminant nutrition section (591 papers-44.3%), followed by the non-ruminant nutrition section (251 papers-18.8%), the animal reproduction section (153 papers-11.5%) and the animal breeding section (115 papers-8.6%). The largest portion of AJAS manuscripts was reviewed by Korean editors (44.3%), followed by Japanese editors (18.1%), Australian editors (6.0%) and Chinese editors (5.6%). Editors from the rest of the AAAP member countries have reviewed slightly less than 5% of the total AJAS manuscripts. It was regrettably noticed that editorial members representing Nepal (66.7%), Mongolia (50.0%), India (35.7%), Pakistan (25.0%), Papua New Guinea (25.0%), Malaysia (22.8%) and New Zealand (21.5%) have failed to return many of the manuscripts requested to be reviewed by the Editor-in-Chief. Financial records show that Korea has contributed the largest portion of production costs (68.5%), followed by Japan (17.3%), China (8.3%), and Australia (3.5%). It was found that 6 AAAP member countries have contributed less than 1% of the total production costs (Bangladesh, India, Indonesia, Malaysia, Papua New Guinea and Thailand), and another 6 AAAP member countries (Mongolia, Nepal and Pakistan, Philippine and Vietnam) have never provided any financial contribution in the form of subscriptions, page charges or reprints. It should be pointed out that most AAAP member countries have published more papers than their financial input with the exception of Korea and China. For example, Japan has published 29.8% of the total papers published in AJAS by AAAP member countries. However, Japan has contributed only 17.3% of total income. Similar trends could also be found in the case of Australia, Bangladesh, India, Indonesia, Malaysia and Thailand. A total of 12 Asian young animal scientists (under 40 years of age) have been awarded the AJAS-Purina Outstanding Research Award which was initiated in 1990 with a donation of US$ 2,000-3,000 by Mr. K. Y. Kim, President of Agribrands Purina Korea Inc. In order to improve the impact factor (citation frequency) and the financial structure of the AJAS, (1) submission of more manuscripts of good quality should be encouraged, (2) subscription rate of all AAAP member countries, especially Category B member countries should be dramatically increased, (3) a page charge policy and reprint ordering system should be applied to all AAAP member countries, and (4) all AAAP countries, especially Category A member countries should share more of the financial burden (advertisement revenue or support from public or private sector).

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.167-176
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• 1997

This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.