• Journal of Aquaculture
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• v.20 no.2
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• pp.96-105
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• 2007

A pair of maroon clownfishes with an indonesian native, reared in recirculation culture system to develope its aquaculture techniques. Courtship, spawning behavior, egg developments and rearing of the maroon clownfish larvae were documented. The larval development were described with illustrative figures. The spawning was occurred 8 times between Feburary and August 2004. The gravid female spawned during 15:00-20:00. The male mainly took care of the eggs supplying oxygen by water currents using their pectoral fins, anal fin and mouth. The fertilized eggs were separative-adhesive and oval in shape, and $1.99{\pm}0.03\;mm$ in longer diameter and $0.88{\pm}0.03\;mm$ in shorter diameter. The fertilized eggs were in deep-orange color. Cleavage occurred in 30 minutes after fertilization, and the egg reached 2 cells stage in 1 hour 10 minutes after fertilization at $27.0^{\circ}{\pm}0.5^{\circ}C$. The embryo was formed in 23 hours 40 minutes after fertilization. Hatching began in between $120{\pm}2$ hours and $150{\pm}12$ hours after fertilization at $27.0^{\circ}C$ in the incubator. Total length (TL) of the newly hatched larvae was 3.22 mm with mouth and anus opened. Ten days after hatching, mean TL of the larvae were 6.21 mm with 28 dorsal fin rays, 17 anal fin rays and 28 caudal fin rays. Nineteen days after hatching, mean TL of the larvae were 9.34 mm. At this stage the larva had three white bands on the body, and they began to feed on commercial diet.

• Applied Microscopy
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• v.20 no.1
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• pp.17-35
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• 1990

Effect of 5-hydroxytryptamine(5-HT, serotonin) and its precursor tryptophan on the cell proliferation of brain and somite parts of 4 day chick embryo in Dulbeco's modified essential medium was examined morphologically at cellular level. It was realized that the externally added 5-HT and/or tryptophan disturbed cell proliferation and severve necrosis occured. Electron micrograph showed that the development of cell organelles were greatly impaired. The activities of both acetylcholine esterase and $Mg^{2+}$ -dependent ATPase of the brain tissues of 5 day chick embryo, which received 1mg of tryptophan and/or 0.1mg of 5-HT at primitive streak stage after 24 hrs incubation of the fertilized egg, were much lower(about 20-25%) than those of control group. These results were supported by the electron micrographs of chemically treated cells. Control cells showed clear densed bands of acetylcholine esterase activity around nucleus and rough endoplasmic reticulum but tryptophan or 5-HT treated groups showed discontinued activity bands. In the case of $Mg^{2+}$-ATPase, the control groups showed clear continuous activity bands but tryptophan and/or 5-HT treated groups were discontinuous. From the previous and present studies, it seems that the intracelluar 5-HT level is very important for the cell proliferation and normal morphogenesis.

• Journal of Korean Ophthalmic Optics Society
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• v.3 no.1
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• pp.9-13
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• 1998

Visual system of Korean Salamander(Hynobius leechi) was morphologically studied. Fertilized eggs in egg sacs were collected and were developed in sterile saline solution. Various sized larvae of 5-30mm in length were fixed. Specimens were paraffin sectioned and were observed under light microscope. In 5mm length larva, lens rudiment induced by optic cup was combined with sensory ectoderm. The shape of lens was changed as spherical in 12mm length larva, but the retinal layer did not differentiated into three layers. The differentiation of retinal layer was clear in 14-16mm length larva. The central region of lens fibers was degenerated. Iris and ciliary body were formed from the marginal zone of optic cup in 20mm length larva. Choroid was thicker in elder eye of 30mm length larva and cartilage developed at outer region of optic cup. The outer segment of photoreceptor cell layer grew longitudinally. Optic nerve was connected to the ventral part of brain through cartilage.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.41 no.6
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• pp.478-484
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• 2008

To elucidate the spawning behavior and early life history of Liobagrus mediadiposalis, mature male and female fish were collected from a branch of the Seomjin River. Spawning was induced by injecting hormones, and then the spawning process and development of fertilized eggs, larvae, and juveniles were observed. Observations of spawning behavior showed that the female established a territory and built a spawning nest, and frequently pressed on the upper ventral part of the male to release her eggs. When spawning was finished, the fish supplied fresh water to the egg mass using their pectoral and caudal fins. Hatching began 189 h 20 min after fertilization at $21.5-23.5^{\circ}C$ (mean $22.7^{\circ}C$). The mean total length (TL) of newly hatched larvae was 7.18-7.39 mm (mean 7.31 mm). Their mouth and anus were already open and they had 14+24=38 myotomes. Eighteen days after hatching, the larvae were 12.71-13.79 mm (mean 13.27 mm) in TL and the yolk sac was absorbed completely. At 35 days after hatching, when all the fin-rays had formed, the juveniles were 15.84-17.92 mm (mean 16.33 mm) in TL.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.12
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• pp.1652-1656
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• 2004

The aim of this study was to obtain mature ova or embryos at a single cell stage, which can be used in avian transgenesis and nuclear transfer through multiple ovulations, in vitro fertilization and culture. Chicken anterior pituitary extract (CAPE) or acetone-dried chicken anterior pituitary extract (ACAPE) was used to induce multiple ovulations in hens pretreated with pregnant mare' serum gonadotrophin (PMSG). In vitro fertilization of the multiple ovulated ova was performed by inseminating sperm onto the germinal disks in m-Ringer' solution and incubating the ova at 41$^{\circ}C$, 5% $CO_2$ for 10 h in DME-F12 medium containing 20% liquid albumen. The in vitro fertilization process was observed using an environmental scanning electron microscope. When normal laying hens (white Leghorn) were administered daily with PMSG (100 IU), egg laying ceased in most hens within 3 to 8 days. Ovulation began to occur about 7.5 h after injection of CAPE and ACAPE. The number of ovulated ova was 1.00${\pm}$0.00, 2.33${\pm}$0.52 and 2.20${\pm}$0.45, respectively, after receiving 100, 200 and 300 mg CAPE. The number of ovulated ova was 2.00${\pm}$0.00, 2.86${\pm}$0.69 and 3.00${\pm}$1.22, respectively, after receiving 10, 15 and 20 mg ACAPE. The fertilized and cultured ova were able to develop into embryos up to the 32 cell stage. The present experiments demonstrated that multiple ovulations can be induced by CAPE and ACAPE successfully, and the ova resulted from the treatment retained the capability for further fertilization and embryonic development. These data provide new information to support the establishment of an in vitro culture system for future avian transgenesis studies.

• Korean Journal of Ichthyology
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• v.32 no.3
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• pp.166-181
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• 2020

The egg development, early life history and spawning site characters of Korean endemic fish, Coreoleuciscus splendidus (Gobioninae), were investigated at the part of Jo-jong stream in Korea from May 2020. The fertilized eggs were 2.05~2.23 mm (mean, 2.13 mm) in diameter and had no oil globules. The embryo began to hatching about 98 hrs after fertilization under water temperature of 20±1℃. The newly-hatched larvae were 5.03~5.68 mm (mean, 5.31 mm) in total length (TL), and their mouth and anus were not opened. 4 days after hatching, Several rod-like cupulae were observed on the head and lateral side of the body at 6.95~7.89 mm (mean, 7.51 mm). 7 days after hatching, the post-larva stage were 8.39~9.29 mm (mean, 8.78 mm) in total length, and their york were completely absorbed. The cupulae were completely distinguished. They entered the juvenile stage when all fin-rays were formed at 29 days after hatching, and their TL were 14.16~17.04 mm (mean, 14.99 mm). Squamation was initiated on the caudal body at approximately 18.21~23.74 mm (mean, 30.28 mm), 38 days after hatching, and completed at 26.82~33.33 mm (mean, 19.42 mm), 73 days after hatching, the external characteristics from of juveniles were same to adults. The spawning site was characterized by bottom structure of pebble (64~16 mm) and gravel (16~2 mm), environmental conditions of the spawning sites were 6~18 cm in water depth, 0.43~0.73 m/sec in bottom water velocity. In spawning sites, an egg mass or separated eggs were located a small gap of under the pebble and gravel.

• Journal of Aquaculture
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• v.17 no.3
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• pp.180-186
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• 2004

This study was carried out to obtain the fundamental data for the mass seedling production of grunt, Hapalogenys nitens in terms of the natural spawning and some characteristics of the eggs spawned. The wild grunt were reared at indoor tanks for three years. The adults spawners were 34.0∼44.0 cm (38.6$\pm$4.0 cm, n=7) in total length, 1.00∼2.23 kg (1.62$\pm$0.50 kg, n=7) in body weight. Spawning were observed 9 times from September 22 to October 1, 2000 and 37 times from August 22 to October 3, 2001, with a water temperature range of 19.8$\pm$28.5$^{\circ}C$. The total number of eggs collected was 2.29${\times}$10$^{7}$ (1.7${\times}$10$^{3}$/ml). The relative proportion of floating eggs to total eggs was 41.7%. The fertilization rate of floating eggs was ranged between 85.0 and 99.9% and the hatching rate was ranged between 2.9 and 93.0%. Fertilized eggs were buoyant and spherical in shape, and were 0.85∼0.98 mm in diameter. Each egg contained 1-5 oil globules which were, 0.18∼0.25 mm in diameter. The incubation time from fertilization to blastodisc formation was 10 minutes, to blastula was 3 hours, and to the hatched larvae at 26$^{\circ}C$ was 20 hours 30 minutes. The newly hatched larvae attained total length of 1.81$\pm$0.18 mm. The time required from fertilization to hatching was 31∼34 hours at 23$^{\circ}C$ and 17∼20 hours at 29$^{\circ}C$.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.1
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• pp.21-28
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• 2002

Objective: To evaluate whether co-culture of oocytes on vero cell monolayers from Day 0 (Day 0 group) after egg retrieval results in an increase in developmental capacity such as fertilization rate, embryo quality, blastulation and clinical pregnancy rate compared with co-culture of oocytes from Day 1 (Day 1 group). Methods: Sperms were treated with Hams F-10 supplemented with 10% human follicular fluid (hFF). Vero cells for co-culture were prepared in TCM-199 with 10% FBS. Oocytes were co-cultured from Day 0 and fertilized oocytes were co-cultured from Day 1 on vero cell monolayers in DMEM with 10% and 20% hFF, respectively after egg retrieval. On day 1, 2 and 5, fertilization rate and grade of embryos and blastocysts were evaluated. Results (fertilization rate, cleavage rate, grade of embryos and blastocysts and pregnancy rate) were considered statistically significant when p value was less than 0.05 using t-test and $x^2$. Results: In sibling oocytes of same cycles, no differences were found in fertilization rate (94.6 vs. 91.4%), cleavage rates (94.6 vs. 91.4%), embryo grade (on day 2 and 3) and blastulation (65.6 vs. 57.0%) and their grade. In different oocytes of different cycles (patients), no differences were found in fertilization (79.8 vs. 78.3%), cleavage rates (77.7 vs. 76.4%) and blastulation (56.0 vs. 45.3%), but pregnancy rate was higher in the Day 0 group than in the Day 1 group (60.0 vs. 42.9%). Conclusions: This study revealed that the embryonic development capacities were not affected by the different co-culture time in the sibling oocytes of same cycles. Although no statistical significance, because of small size of study, there was a trend for higher pregnancy rates in Day 0 group compared to Day 1 group in different oocytes of different cycles.

• Development and Reproduction
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• v.20 no.2
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• pp.119-125
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• 2016

This study describes results on sexual maturation and characteristics of natural spawned eggs to develop a method for the production of stable, healthy fertilized eggs from captive-reared yellowtail kingfish, Seriola lalandi. A total of 59 yellowtail kingfish were captured off the coast of Jeju Island, after which the broodstock was cultured in indoor culture tank ($100m^3$) until they were 6.1-14.9 kg in body weight. As part of the rearing management for induced sex maturation, the intensity of illumination was maintained at 130 lux. The photoperiod (light/dark; L/D) was set to a 12 L/12 D from October 2013 to January 2014, and 15 L/9 D from February 2014 to June 2014. Feeds comprised mainly EP (Extruded Pellets), with squid cuttlefish added for improvement of egg quality, and was given from April to June 2014. The first spawning of yellowtail kingfish occurred in May 3, 2014, at a water temperature of $17.0^{\circ}C$. Spawning continued until June 12, 2014, with the water temperature set at $20.5^{\circ}C$. Time of spawning was 26 times at this period. The total number of eggs that spawned during the spawning period was $4,449{\times}10^3$. The buoyant rate of spawning eggs and fertilization rate of buoyant eggs during the spawned period were 76.1% and 100%, respectively. The diameters of the egg and oil globule were $1.388{\pm}0.041mm$ and $0.378{\pm}0.029mm$, respectively, which was higher in early eggs than in those from late during the spawned period.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2001.11a
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• pp.74-76
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• 2001

Comparing to mammals, male bird has the homozygote ZZ and female has the heterozygote n. Therefore, the sex of fertilized eggs is defined by female chromosome constitution. Although this cytological observation had been established, the molecular and cellular mechanism of germ cell differentiation are essentially unknown in aves. Especially, the differentiation of germ cells in mixed-sex chimeras has not yet been clearly elucidated. Primordial germ cells, which are the progenitors of sperm or egg after sexual maturity, firstly arise in the epiblast and migrate to embryonic gonads through the blood vessel. During the embryo development, these PGCs differentiate in the pathway of mate or female, respectively and develop the sperm or egg cells after sexual maturity. In this paper, we confirmed that the female PGCs could migrate into the recipient male gonads after transferring and differentiate into germ cells in the embryonic stages. The primordial germ cells were isolated from the female embryonic gonads of 5.5-day-old incubation and re-injected into the male recipient embryos of 2-day-old incubation, which produced mixed-sex chimera in the germline. The finding in this study demonstrated the ability of migration and differentiation of gonadal primordial germ cells in mixed-sex chicken.