• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.90-90
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• 2002

The purpose of this study was to investigate the effect of kind and combination of sugars on the viability and acrosome damage of post-thaw spermatozoa in canine. The extender used was Tris-citric acid extender (Tris-buffer) supplemented with 20% Egg-yolk, 8% glycerol, 1% Equex STM paste, and 70 mM sugars such as monosaccharide (fructose and xylose) and disaccharide(trehalose). To evaluate of sugar combination, the sugars supplemented in Tris-buffer were combined such as single (fructose, xylose, trehalose), two combinations (Fruc+Tre, Fruc+xyl, Tre+xyl) and three combinations (Fruc+Tre+Xyl). The concentration of sperm collected were adjusted of 50${\times}$10$\^$6/ per straw for freezing. The frozen spermatozoa were thawed at 37$^{\circ}C$ for 1 min and then analysis for CASA program in Livestock Improvement Main Center, NACF. The motility of post-thaw spermatozoa in Fruc+Tre was higher than those in fructose, trehalose, xylose, Fru+xyl, Tre+xyl and Fru+Tre+Xyl (79％ vs. 63, 66, 70, 71, 74 and 75%). The progressive motility after CASA analysis in Fuc+Tre group was also higher than those in fructose, trehalose, xylose, Fru+xyl, Tre+xyl and Fru+Tre+Xyl (67% vs. 53, 57, 60, 61, 62 and 64%). The acrosome damage of post-thaw spermatozoa stained was not significantly different among treatment groups such as fructose, trehalose, xylose, Fru+Tre, Fru+xyl, Tre+xyl and Freu+tre+xyl (17.7, 18.3, 28.0, 17.0, 19.7, 20.0 and 19.0%). The results indicated that the motility and progressive motility of post-thaw spermatozoa in Fru+Tre group was better, and acrosome normality was not different among all groups. The use of Tris-buffer supplemented with Fru+Tre as sugar for frezing of canine spermatosoa could be better and apply to semen banking and artificial insemination.

• Korean Journal of Ichthyology
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• v.13 no.1
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• pp.74-84
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• 2001

The early developmental stages, growth and morphological changes of the Korean bullhead, Pseudobagrus fulvidraco, were studied from a series of reared specimens. Details of the early developmental stages are illustrated with special reference to morphological transformations. Egg and sperm of Korean bullhead were obtained from mature adults under hormonal treatment, fertilized artificially, and incubated in the aquarium. The incubation period of fertilized eggs was 55 to 66 hours at a temperature of 24.9${\pm}$0.34$^{\circ}$. Larvae were fed successively with Artemia salina and Daphnia magna for 2 to 15 days and artificial food after 20 days. Fertilized eggs were adhesive and spherical with a diameter of 2.04mm(n = 100). The mean total length of newly hatched larvae was about 4.92${\pm}$0.33 mm. Mouth opening occurred on one-day-old yolk-sac larvae, and initial feeding was observed on the third day after hatching. The morphological transitions from larvae to juvenile and juvenile to young stages occurred when the fish reached about 17 mm in total length (about 13days after hatching) and about 32 mm in total length (about 30 days after hatching), respectively. Many changes in proportion of body parts to total length were observed at about 7~8 mm and 30~32 mm, corresponding to the transformations from larvae to juvenile and from juvenile to young, respectively. In comparing relative growth of each body part against total length, those characteristics related to head parts showed positive growth in the pre-larval stages, while those concerning mobile abilities showed positive growth in the post-larval stage.

• Korean Journal of Veterinary Research
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• v.30 no.4
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• pp.361-371
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• 1990

The present observations were focussed mainly on the morphological findings of the utero-vaginal(U-V) glands in normal laying domestic hens and the storage of cock spermatozoa in the U-V glands at various times after artificial insemination(AI). These domestic hens were assigned to three group of PMS-treated, GnRH-treated before last AI, and control group. The hens were sacrified at intervals of 1,3,7,12 and 19 days after AI. Histological sections of U-V junctions were prepared and the morphological structures of the U-V glands were observed and then were scored about the spermatozoa presence in the U-V gland. 1. The U-V glandular tubules were mostly unbranched with single columnar epithelium. Also these tubules were occassionally observed as one circular-rotated tubules or 2 to 3 branched convoluted tubules in special shapes. The numbers of the convoluted curves per tubule were $4.3{\pm}3.3$ and the ranges of convoluted curve number were straight to 16 curves. 2. The inside and outside diameters of the glandular tubules were $6.5{\pm}3.5{\mu}m$, and $35.2{\pm}4.7{\mu}m$, respectively, and the tubular lengths of the U-V glands were $219.3{\pm}115.7{\mu}m$. 3. Storaged spermatozoa in the U-V glands of all three group hens were intensively stained by hematoxylin, and packed in tight, longitudinally parallel bundles within the tubules. In addition, numbers of completely spermatozoa-filled glands were tend to increase or decrease in proportion to the numbers of partially spermatozoa-filled glands. Also U-V glands containing spermatozoa tend to be present collectively in the any zone of U-V junction. 4. In the control group, the numbers of glands containing spermatozoa in the hens at 1,3,7,12, and 19 days after AI were found to be 22.9, 33.3, 35.8, 8.6, and 0% respectively. 5. In the PMS-treated group, the numbers of glands containing spermatozoa in the hens at 1,3,7,12, and 19 days after AI were found to be 33.6, 29.7, 26.8, 8.2 and 0% respectively. 6. In the GnRH-treated group, the numbers of glands containing spermatozoa in the hens at 1,3,7,12, and 19 days after AI were found to be 19.7, 40.8, 20.4, and 0% respectively.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.193-201
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• 1999

In this study we determined fertilization processes and developmental ability of porcine oocytes following injection of round spermatid in the presence of artificial activation. Electrical stimulation at 3 h before spermatid injection significantly increased the incidence of normal fertilization as compared to those following injection without stimulation or with stimulation immediately after injection. The incidences of two pronuclear formation and apposition were not different in oocytes between following intracytoplasmic spermatid and spermatid nucleus injection. Indirect immunocytochemistry and laser scanning confocal microscopy study revealed that micro tubules were organized from the oocyte cortex following round spermatid injection, and this seemed to move both male and female pronuclei into the oocyte center. Paternal mitochondria which are introduced with spermatid have been observed up to 4-cell. Our study indicated that either round spermatid or it's nucleus can be used to produce viable bovine embryos by injection into unfertilized oocytes.

• Journal of Aquaculture
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• v.10 no.1
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• pp.25-32
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• 1997

Develoment precess and characteristics of eggs of the geoduck clam, Panope japonica are reporting in this study. Eggs and sperm were excised from gonad, artificially fertilized in an aquarium, reared under various temperature regimes, and record and record the larval period and the time need to reach a certain larval stage from ferilization. Unfertilized eggs of P. japonica appeared to be oval with a mean diameter of $70\mu$m and they became spherical after fertilization. The eggs of P. japonica can be classified as demersal. At a constant water temperature of $ 11^{citc}C$, it took 4 hours form fertilization to become four-cell stage, two days to become trochophore larvae, three days to become D-shape larvae, twenty-three days to become umbo stage, and thirty-six days to become fully grown veliger ready form settlement. A negative correlation was observed between the water temperature and the larval period of P. japonica. From fertilization to D-shape larvae, it took five days at 8$^{\circ}C$, while it was only two days to become D-shape larvae at $ 17^{citc}C$. Time required to D-shape larvae from fertilization was proportional to temperature, and the relationships were expressed as follows : To 8-cell stage, 1/t=0.0209 w-0.1167 (r=0.9967) To blastula stage, 1/t=0, 0055 w-0.0192 (r=0.9825) To trochophore stage, 1/t=0.0034 w-0.0155 (r=0.9907) To D-shape larvae stage, 1/t=0.0014 w-0.0023 (r=0.9843) (t, time in hours ; w, water temperature) Bioligical minimum temperature for egg development was calculated as 3.82$^{\circ}C$ in average.

• Korean Journal of Ichthyology
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• v.29 no.2
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• pp.101-108
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• 2017

Egg development and early life history of the Korean natural monument fish Hemibarbus mylodon (Pisces: Cyprinidae) were investigated to provide basic data on biological characteristics and ecological recovery. Adult fish were collected from nature and transferred to the laboratory. For the first time, artificial maturation of females and males succeeded after 15 months of indoor culture. Mature eggs and sperm were obtained using Ovaprim injections (0.5 mL/kg) and then the eggs were fertilized using the dry method in the laboratory. The mature eggs were adhesive, turbid, and greyish; they averaged $2.21{\pm}0.06mm$ (n=30) in diameter. The embryos began to hatch about 78 h after fertilization at a water temperature $20{\pm}1^{\circ}C$, and the newly-hatched larvae were $6.6{\pm}0.75mm$ in total length (TL). At 14 days after hatching, the post-larval individuals were $13.5{\pm}0.23mm$ (TL), and their yolk sacs were completely absorbed. Twenty one days after hatching, they entered the juvenile stage and reached $13.5{\pm}0.23mm$ (TL). At 100 days after hatching, their band patterns, external form, and a pair of barbels were similar to those of adults, and they averaged $33.0{\pm}4.25mm$ (TL). The breeding technology and characteristics of early life history obtained in this study will be very helpful for conservation of H. mylodon in nature.

• Korean Journal of Medicinal Crop Science
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• v.2 no.1
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• pp.81-85
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• 1994

This experiment was carried out to investigate the fertilization and the size of mature female and male gametophytic parts of Pulsatilla koreana after artificial pollination. The size of pollen is $26.5{\mu}m$ at the time of anther dehiscence, that is, about $3{\sim}4$ days after pollination. Synergid nucleus, egg nucleus, and polar nucleus are 10.0, 15.0, and $32.5{\mu}m$ respectively at the time of completing egg apparatus formation, that is, about 2 days after pollination. Poller tubes germinate on stigma about 10 hours, passing lower part of style about 30 hours, penetrating into micropyle about 35 hours after pollination. Sperm nucleus penetrates into polar nucleus about 40 hours and egg cell about 48 hours after pollination. But, there seems to be different among the individuals. Multinuclei and multinucleoli are formed in egg cell, synergid, and polar nucleus about the time of fertilization. Proembryo is formed about 4 days, being changed to large globular form about $6{\sim}8$ days after pollination. Endosperm nuclei divide into free nuclei after fertilization and change to cotyledon in gymnosperm. There seems to be same phenomena in Pulsatilla koreana.

• Korean Journal of Poultry Science
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• v.41 no.1
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• pp.21-27
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• 2014

To preserve chicken genetic materials like cryopreserved spermatozoa, various kinds of freezing agents like glycerol, dimethylsuloxide, dimethylformamide or dimethylacetamide have been used for rooster semen preparation. Recently, the usage of N-methylacetamide (MA) for Ogye rooster semen preservation resulted in hatched chicken successfully. In this study, we investigated the effects of 7, 9 and 11% of MA on the viability, fertility and hatchability of frozen-thawed rooster semen using artificial insemination. The results of viability, fertility and hatchability in frozen semen with 7%, 9% or 11% MA were $35.16{\pm}6.12%$, $67.83{\pm}15.3%$ and $66.2{\pm}16.3%$ of motile sperm rate, 21.5%, 34.7% and 25% of fertility rate, and 100%, 89.5% and 87.5% of hatchability rate. The results of control group with frozen semen were 96.0% of fertility rate and 92.2% of hatchability rate. With these results, the concentration range of MA as a freezing agent of rooster semen could be 7~9% of media. The higher concentration of 9 % MA could decrease the fertility rate of thawed semen not the rate of hatchability rate. So the use of MA without affecting fertility rate would be a key point of freezing method of rooster semen for poultry genetic resource preservation.

• Korean Journal of Poultry Science
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• v.39 no.4
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• pp.291-295
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• 2012

The use of methylacetamide (MA) as a cryoprotective agent for freezing Korean Native Black rooster Ogye semen was examined with artificial insemination. The diluted Ogye semen with HS-1 was subjected for 2 step dilution method of cryopreservation in which the final concentration of MA was adjusted to 7.5%. The sperm viability after thawing was reduced from $95.17{\pm}0.93%$ to $55.93{\pm}1.38%$ which was confirmed by live-death analysis based on Fluorescence-Activated Cell Sorting (FACS). The rates of fertilized eggs with fresh or frozen-thawed semen were reduced from $94.98{\pm}3.93%$ to $66.36{\pm}8.43%$ at day 7 with significant difference. However, the hatching rates of experiments at day 21 did not shown difference between $92.64{\pm}2.33%$ and $90.45{\pm}8.05%$ (P<0.05). With these results, the utilization of MA for freezing of Ogye spermatozoa could affect on viability of frozen-thawed semen but not on the fertility of lain eggs and hatchability of fertilized eggs and also provide possible tools of freezing for poultry genetic resource conservation.