• Fisheries and Aquatic Sciences
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• v.16 no.1
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• pp.15-23
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• 2013

Normal embryonic development at a constant temperature ($18^{\circ}C$) has been described for the Siberian sturgeon Acipenser baerii (Acipenseriformes). Hormone-induced spawning and artificial insemination were performed to prepare embryonic batches for embryologic examination. After insemination, early cleavages of the Siberian sturgeon embryos continued for 7 h post-fertilization (HPF), showing the typical pattern of uneven holoblastic cleavage. Blastulation and gastrulation began at 9 HPF and 19 HPF, respectively. Epiboly formation (2/3 covered) was observed at 25 HPF during gastrulation. Neurulation was initiated with the formation of a slit-like neural groove from the blastopore at 33 HPF. During neurulation, the primary embryonic kidney (pronephros) and s-shaped heart developed. The embryos underwent progressive differentiation, which is typical of Acipenseriform species. A mass hatching was observed at 130 HPF, and the average total length of the hatched prolarvae was 10.5 mm. The hatched prolarvae possessed a typical pigment plug (yolk plug). The results of this study are valuable not only as a reference guide for the artificial propagation of Siberian sturgeon in hatcheries but also as the basis for the derivation of developmental gene expression assays for this species.

• The Korean Journal of Malacology
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• v.30 no.2
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• pp.127-134
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• 2014

Gonad maturation of Manila clam, Ruditapes philippinarum was induced in this study using a recirculation system over 8 weeks in early spring. Clams used in the experiment were collected in $15^{th}$ April 2010 from the west coast of Korea, as the surface water temperature remained $11^{\circ}C$. To induce gametogenesis and subsequent maturation seawater temperature was elevated $1^{\circ}C$ per day over 10 days to reach $20^{\circ}C$. For the experiment, clams were raised in 120 L quadrangle tank maintained with re-circulated seawater system over 57 days. Water quality parameters including the water temperature, salinity dissolved oxygen, ammonium ion and nitrate levels in the tanks were monitored daily. Mixture of concentrated microalgae including Tetraselmis, Isochrysis, Pavlova and Thalassiosira weissflogii was supplied to clams twice a day, and quantity of the daily ration was adjusted as 3% of clam body dry weight. Histology was applied to examine gonad maturation. Daily monitoring of the water quality parameters indicated that the recirculation system supplied suitable environment to Manila clam; the nitrogenous components stayed below toxic levels (< 0.2 mg/L). At the beginning of the study, clams were mostly in early developing stage. As the seawater temperature reached $20^{\circ}C$, 10 days after the experiment, 20% of clams reached late development at 12 days. First ripe clams were observed at 42 days and 40% of clams were in ripe and ready for spawning at the end of study, 57 days after the experiment. In this study, gametogenesis of Manila clam was successfully induced by elevating water temperature and supplying commercially produced microalgae in a recirculation tank system.

• Journal of Aquaculture
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• v.9 no.1
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• pp.3-10
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• 1996

Ovulation of maturing female yellow puffer, Takifugu obscrus, was induced by using single injection of human chorionic gonadotropin (HCG) or gonadotropin releasing hormone-analogue (GnRH-A) $des-Gly^{10}[D-Ala^6]$ GnRH-ethylamide plus pimozide. The response was evaluated using the fertilization and embryo-formation rate after insemination and the gonadotropin (GTH) level in blood plasma using radioimmunoassay. In the fertilization and embryo-formation, maximal effects were recorded by using 1,000 IU/kg HCG or $10\;{mu}g/kg$ GnRH-A plus 5 mg/kr pimozide. Pimozide (1, 5 mg/kg) or GnRH-A treatment alone was not effective in elevation of GTH level, however combinations of these treatments were particularly effective. Injection of dopamine blocked the rapid elevation of plasma GTH levels of blood. These data suggest that yellow puffer secrete GnRH and gonadotropin-releasing-inhibiting factor during the spawning or the other period.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.1
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• pp.41-47
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• 2017

Perhaps the most common type of reproductive dysfunction in captive fish is failure of females to undergo final oocyte maturation and thus to ovulate and spawn. The success of aquaculture could therefore be improved by developing techniques to enhance natural spawning, artificial maturation, and/or to induce ovulation in farmed fish. This study aimed to investigate the effects of prostaglandin $E_2$ ($PGE_2$) and prostaglandin $F_{2{\alpha}}$ ($PGF_{2{\alpha}}$) on in vitro oocyte maturation (germinal vesicle breakdown, GVBD) and ovulation in the marine fish Chasmichthys dolichognathus. Post-vitellogenic follicles (0.80-0.94 mm diameter oocytes) were incubated with $PGE_2$ or $PGF_{2{\alpha}}$ at concentrations of 5, 50, or 500 ng/mL for 24 hours. A significant increase in GVBD was seen in 0.84 mm and 0.94 mm oocytes incubated with 50 ng/mL $PGE_2$ compared with the control. There was no significant increase in GVBD in any of the other experimental conditions (5 or 500 ng/mL $PGE_2$ or 5, 50, or 500 ng/mL $PGF_{2{\alpha}}$). Neither of the prostaglandins induced ovulation at the concentrations tested.These results suggest that GVBD was induced by incubation with 50 ng/mL $PGE_2$.

• Journal of Aquaculture
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• v.20 no.3
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• pp.160-167
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• 2007

Embryonic and pre-leptocephalic larvae development of the eel, Anguilla japonica, are described following natural fertilization in the indoor tank of $23^{\circ}C$ water temperature. Following a routine hormone treatment technique for the brood stock, female eels were artificially matured by weekly intramuscular injections of salmon pituitary extracts (SPE) at a dosage of 20 mg/kg body weight (BW) for a total of 10-14 doses to induce ovarian maturation, while male eels received weekly intramuscular injections of human chorionic gonadotropin (HCG) at a dosage of 1 IU/g BW for a total of 6-10 doses to induce testicular maturation in a separate aquarium and induced natural spawning. Fertilized eggs of about 1.0 mm in diameter were pelagic and showed a typical discoidal cleavage. Hatching occurs 38 hrs after fertilization at a water temperature of $23^{\circ}C$. The newly hatched larvae measured about 3.0 mm in total length and the number of myomeres averages 42. Their mouths and anuses were opened at 4.5 days and the yolk sacs of the pre-leptocephalic larvae were almost absorbed at 6.5 days after hatching. Pre-leptocephalic larvae survive for 14.5 days. At this time they are $5.87{\pm}0.25mm$ in total length and have about 98 myomeres. However, morphological characterization of embryonic and pre-leptocephalic larvae were not different between natural fertilization and artificial fertilization by the dry method.

• Korean Journal of Ichthyology
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• v.26 no.2
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• pp.89-98
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• 2014

Eggs development and early life history of the endangered Korean dwarf loach, Kichulchoia brevifasciata (Teleostei: Cobitidae) was investigated to provide basic information regarding biological characteristics and restoration. Adult fish specimens were sampled using a spoon net at Geurnsan-myeon, Goheung-gun, Jeollanam-do, Korea from June to July 2011. Since, spawning characteristics were analyzed, and females were induced to spawn by injecting Ovaprim (0.5 mL/kg) and their eggs were artificially fertilized with sperms by the dry method in the laboratory. Total length of mature female were 46~76 mm with GSI $9.6{\pm}3.77%$, and total length of mature male was 42~52 mm with GSI $3.5{\pm}1.04%$. Sex ratio (♂/♀) was 0.10, and there were no secondary sexual characteristics. The number of mature eggs was averaged $60{\pm}28.7$ per female. The lemon yellow eggs were slightly adhesive $1.46{\pm}0.07mm$ in diameter. The embryo hatched approximately 66 h after fertilization at $25^{\circ}C$, and the hatched larvae were averaged $5.5{\pm}0.07mm$ in total length (TL). At 6 days after hatching, the larvae averaged $9.0{\pm}0.29mm$(TL) and their yolk sac was completely absorbed. At 17 days after hatching, they entered the juvenile stage and reached $12.6{\pm}0.24mm$ (TL). At 80 days after hatching, the band patterns and external form of the juveniles were similar to those of adults, and they averaged $33.0{\pm}2.19mm$(TL).

• Environmental Analysis Health and Toxicology
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• v.25 no.1
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• pp.11-18
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• 2010

Toxicity assessment of ocean dumping wastes (dye waste, urban sewage, food waste) were examined in the fertilization and embryo development rates of the Sea Urchin, Hemicentrotus pulcherrimus. Spawning was induced by injecting 1 mL of 0.5 M KCl into coelomic cavity. Males released white or cream-colored sperms and females released yellow or orange-colored eggs. Experiments were began within 30 min the collection of both gametes. The fertilization and embryo development rates test were performed for 10 min and 64 h after fertilization, respectively. The fertilization and embryo development rates in the control condition (not including ocean dumping wastes sludge elutriate) were greater than 90%, but suddenly decreased with increasing of ocean dumping waste sludge elutriate concentrations. The fertilization and normal embryogenesis rates were significantly inhibited in all waste sludge elutriate from dye waste ($EC_{50}$=4.37; $EC_{50}$=1.76), urban sewage ($EC_{50}$=5.79; $EC_{50}$=2.00) and food waste ($EC_{50}$=7.68; $EC_{50}$=2.16), respectively. The NOEC (<3.13) and LOEC (3.13) of fertiliztion and normal embryogenesis rates very similar in all waste sludge elutriate. These results suggest that biological assay using the fertilization and embryo development rates of H. pulcherrimus are very useful test method for the ecological toxicity assessment of ocean dumping wastes.

• Journal of Environmental Science International
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• v.17 no.7
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• pp.775-781
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• 2008

Effects of salinity and standard toxic metals on fertilization and embryo development rates were investigated in the sea urchin, Strongylocentrotus nudus. Spawning was induced by injecting 1ml of 0.5 M KCl into the coelomic cavity. The fertilization and embryo development rates were below 20% when salinity was 25 psu or lower, but were above 90% when salinity was between 30 and 35 psu. The fertilization and embryo development rates in the control condition (not including Cu and Cd) were greater than 90%, but decreased with a high negative correlation (r) of 0.89 and 0.91 with the increasing of Cu and Cd concentrations, respectively. These results suggest that salinity concentrations for successful fertilization and normal embryogenesis of S. nudus are between 30 and 35 psu, and the biological assays of fertilization and embryo development rates using S. nudus are useful methods for the ecological toxicity test of marine pollution elements.

• Journal of Aquaculture
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• v.13 no.1
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• pp.47-53
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• 2000

Hormone induction in maturation and ovulation was carried out with spotted halibut (Verasper variegatus) which normally does not spawn spontaneously in captivity. Prior to spawning females were treated with human chorionic gonadotropin(HCG 265-678 IU/kg BM) 17 ${\alpha}$-hydroxy 20-${\beta}$-dihydroprogesterone (17 ${\alpha}$ 20${\beta}$ OHP 0.5-1.0${\mu}$g/kg BW) and luteinizing hormone-releasing hormone analogue pellet (LHRHa pellet 63-81 ${\mu}$g/kg BW) In all the trials HCG injection succeeded in inducing ovulation at 265-300 IU/kg BW. HCG at 620 or 678 IU/kg BW was ineffective in inducing a good rate of ovulation. Final maturation and ovulation were not stimulated by treatment of 17 ${\alpha}$ 20${\beta}$OHP and LHRHa pellet. These results suggest that HCG injection (around 300 IU/kg BW) is more effective compared with LHRHa pellet implantation.

• Environmental Analysis Health and Toxicology
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• v.24 no.1
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• pp.9-16
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• 2009

Effects of salinity and standard toxic metals on fertilization and embryo development rates were investigated in the sea urchin, Hemicentrotus pulcherrimus. Spawning was induced by injecting $1{\sim}2$ mL of 0.5 M KCl into the coelomic cavity. Males released white or cream-colored sperms and females released yellow or orange-colored eggs. The fertilization rate was below 30% when salinity was 20 psu and lower, but was almost above 90% when salinity was 25 psu and higher. The embryo development rate was below 60% when salinity was 25 psu and lower, but was above 80% when salinity was between 30 and 35 psu. The fertilization and embryo development rates in the control condition(not including Cu and Cd) were greater than 90%, but decreased a high negative correlation with the increasing of Cu(r=-0.80, r=-0.78) and Cd(r=-0.90, r=-0.82) concentrations, respectively. The fertilization and embryo development rates were significantly inhibited in the addition of Cu($EC_{50}$=17.30 ppb, $EC_{50}$=10.32 ppb) and Cd($EC_{50}$=364.57 ppb, $EC_{50}$=244.04 ppb), respectively. These results suggest that salinity concentrations for successful fertilization and normal embryogenesis of H. pulcherrimus are above 25 psu and 30 psu, respectively, and the biological assays of fertilization and embryo development rates using H. pulcherrimus are useful methods for the ecological toxicity test of marine pollution elements.