• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.72-72
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• 2003

The present study examined the possibility of cryopreservation of the D-shaped and umbo larvae of arkshell (Scapharca broughtonii), in terms of the survival rates after freezing and thawing. D-shaped and umbo larvae of arkshells were obtained from a shellfish farming on Yosu city. The average shell lengths were $93.3 \pm 10.1 \mu$m and $201.7 \pm 13.5 \mu$, respectively. Five cryoprotectants (CPAs), dimethyl sulfoxide (DMSO), glycerol, ethylene glycol (EG), propylene glycol (PG), and methanol, were tested at the concentrations of 1.5, 2.0 and 2.5 M. After larvae suspended in CPAs, cryoprotectants were loaded in 0.5 ml straws at a larval density of 50-100 larvae per straw, and epuilibrated for 10 and 20 minute at room temperature ($23^{\circ}C$), repectively. Straws were cooled at a rate of $1^{\circ}C$/min from $0^{\circ}C$ to $-12^{\circ}C$, held for 5 min at $-12^{\circ}C$, and then cooled at $2^{\circ}C$/min to $-35^{\circ}C$ and equilibrated for 5 min followed by plunging in liquid nitrogen. After storage in liquid nitrogen for 1 day, straws were thawed in a $30^{\circ}C$ water. As soon as straws were observed to melt, larvae were diluted with an equal volume of ASW and then washed twice with a large volume of ASW at an interval of 2 min to unload the CPAs. The results showed that after equilibration for 10 and 20 minute at room temperature, no larvae survived using methanol as CPAs, and it was observed that larval shells all open slightly, and larval flesh broke down and slopped over the shells. The highest survival rates (D-shaped larvae: 77.6%, umbo larvae: 59.3%) were obtained with 2M DMSO, and 1.5M glycerol yielded survival rates of 53.8% for D-shaped larvae and 37.5% for umbo larvae. The surviving D-shaped larvae showed active rotary motion and perfect membrane integrity and cytoplasmic normality, and the vigorous movement of veliger cilia was observed inside the closed shells. The breakdown of tissue occurred in the abnormal larvae, and the isolated cell often run out of shells.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.7 no.1
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• pp.1-6
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• 1974

Meretrix lusoria is one of the most favorite edible bivalves inhabiting wide inter-tidal flats along the western coast of Korea. Over the period of July to September 1973, some specimens from a tidal flat near AnMyun Island were submerged in sea water with various concentrations of ammo-mum hydroxide added and careful observations were made on their fertilization, early development, and metamorphosis of the larvae. The highest rate of fertilization was demonstrated by individuals treated with 1/1000 normal solution of ammonium hydroxide, and their fertilized eggs followed normal development, i. e., two cell stage in 1.2 hours after fertilization, gastrula stage after 4.7 hours, and trochophore stage after 5.6 hours. Within 24 hours after fertilization M. lusoria larvae have acquired the form of early straight-hinge veliger with the mean prodissoconch I length of $112\mu$. It takes seven days to get the umbo stage with the mean shell length of $172\mu$ and twenty days to get the metamorphosing stage with the mean shell length of $232\mu$. The larvae were cultured to the metamorphosing stage with the shell length of $272\mu$ in the laboratory condition. The relationship between the shell length (L) and the shell height (H) in veliger stage is shown as H=1.02325L-24.46425 with a significant difference.

• Animal cells and systems
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• v.3 no.4
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• pp.375-383
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• 1999

Gonadal development, gametogenesis, reproductive cycle, and first sexual maturity of Reishia clavigera were investigated monthly from July 1998 to June 1999 through cytological and histological observations. R. clavigera had separate sexes, and was an internal fertilizer. The ma1e penis was located near the two tentacles. The ovary and testis were composed of a great number of oogenic lobules and spermatogenic tubules, respectively. The size of ripe oocyte ranged from 130 to 140 ${\mu}$m in diameter. The peripheral cytoplasm of the germinal vesicle of the ripe oocyte in many cases were surrounded by smaller yolk granules, while the eccentric cytoplasm was occupied with larger ones. The reproductive cycle of R. clavigera could be classified into five successive stages: early active, late active, ripe, spawning, and recovery. Spawning of females occurred from early July to August when the seawater reached above 24.8$^{\circ}C$. Spawning of males occurred from early June to August in the water above 22.8$^{\circ}C$. Minimum size for sexual maturity of both sexes was above 10.0 mm in shell height. Each egg capsule was a cylinder or spindle in shape, 4-6 mm in length and 1-2 mm in width. Colors of newly spawned egg capsules showed yellowish white or pale yellow, while those with veliger larvae showed pale black, and released larvae or dead egg capsules showed black violet. The fecundity in an egg capsule ranged from 70 to 91 eggs (mean＝80.28 eggs).

• 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 Fisheries and Aquatic Sciences
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• v.34 no.2
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• pp.131-136
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• 2001

Relative growth and composition of larval development stages in brood pouch of the melania snail, Semisulcospira libertina libertina were investigated based on monthly field samples for a year. Total number of specimens examined was 1,200. Size values (mean$\pm$S. D.) of shell height (SH), shell diameter (SD), total weight (TW), meat weight (MW) and shell weight (SW) were $15.96\pm3.41\;mm$, $7.82\pm1.38\;mm$, $0.55\pm0.31\;g$, $0.24\pm0.15\;g$ and $0.39\pm0.23\;g$, respectively, Water temperature (WT) of the sampling site varied from $1.3^{\circ}C\;to\;22.5^{\circ}C$. Relative growth equations among SH, SD, TW, MW and SW of S. libertina libertina specimens pooled were as follows: SD=0,3583SH+2.1648 $(R^2=0.8391)$, $TW=0.0005SH^{2.4853}\;(R^2=0.8391)$,$MW:\;0.0005SH^{2.849}\;(R^2= 0.8391)$ and $SW=0.0004SH^{2.3798}\;(R^2=0.7057)$. Female S. libertina libertina had larvae in its brood pouch throughout the year, Average number of larvae in the brood pouch varied from $91\pm33(July)$ to $396\pm174\;(November)$, decreasing in spring and increasing in autumn. In the monthly composition of larval stage in the brood pouch, trochophore, pre-veliger, veliger and juvenile showed maximum as $66.7\%$ in May, $70.7\%$ in November, $60.9\%$ in February and $13.9\%$ in January, respectively. Percent composition of the juveniles decreased in spring and autumn. In conclusion, S. libertina libertina is likely to have two main parturition periods, March to May and September and October.

• The Korean Journal of Malacology
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• v.29 no.2
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• pp.91-96
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• 2013

Although the method of seedling production of Haliotis discus hannai is well known, the optimum benthic diatom species as a live food at early larval stage are not fully developed. In this study three Pennales diatom species, Caloneis schroederi, Rhaphoneis sp., and Cocconeis californica were examined on settlement, metamorphosis, survival, and growth of Haliotis discus hannai larvae. The larvae fed Raphoneis sp. or C. californica showed high settlement rate with 80-82% within 48 hrs, which was significantly higher than those fed C. schroederi or mixed diets with three diatom species. The larvae fed the former microalgal species also showed higher metamorphosis rate with 32-34% than the latter species with 10-12% within 4 days. With regard to survival and growth of the larvae, single diet with Rhaphoneis sp. or C. californica had better dietary value than the mixed diets for the early larvae of H. discus hannai.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.36 no.6
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• pp.625-631
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• 2003

The aim of the present study was to investigate the ecological relationship between female body size (=shell length) and fecundity in a Korean population of protandric hermaphrodite, Crepidula onyx Sowerby, using 6 reproductive variables (the number of capsules per brood, capsule size, the number of embryos per capsule, total number of embryos per brood, egg size, and larval size). The investigation was based on 32 females, 107 egg capsules, 263 eggs, and 250 veliger larvae sampled in January 2000, from Yangpo, the southeastern coast of Korea. All foregoing reproductive variables, except larval size, were significantly correlated with the female body size (p<0.001$\~0.01$). There was a significant increase both in the number of capsules per brood and the number of embryos per capsule with female body size (p<0.001). Consequently, the fecundity of the female C. onyx increased with female body size to over 14.65 mm (the minimum size of egg brooding in the present study). This investigation, therefore, provides additional evidence that female fecundity of protandric hermaphrodites is positively correlated with female body size.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.3 no.3
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• pp.177-186
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• 1970

The spawning of the abalone, Haliotis discus hannai, was induced In October 1969 by air ex-position for about 30 minutes. At temperatures of from 14.0 to $18.8^{\circ}C$, the youngest trochophore stage was reached within 22 hours after the egg was laid. The trochophore was transformed into the veliger stage within 34 hours after fertilization. For $7\~9$ days after oviposition the veliger floated in sea water and then settled to the bottom. The peristomal shell was secreted along the outer lip of the aperture of the larval shell, and the first respiratory pore appears at about 110 days after fertilization. The shell attained a length of 0.40 mm in 15 days, 1.39 mm in 49 days, 2.14 mm in 110 days, 5.20 mm in 170 days and 10.00 mm in 228 days respectively. Monthly growth rate of the shell length is expressed by the following equation :$L=0.9981\;e^{0.18659M}$ where L is shell length and M is time in month. The density of floating larvae in the culture tank was about 10 larvae per 100 co. The number of larvae attached to a polyethylene collector ($30\times20\;cm$) ranged from 10 to 600. Mortality of the settled larvae on the polyethylene collector was about $87.0\%$ during 170 days following settlement. The culture of Nauicula sp. was made with rough polyethylene collectors hung at three different depths, namely 5 cm, 45 cm and 85 cm. At each depth the highest cell concentration appeared after $15\~17$ days, and the numbers of cells are shown as follows: $$5\;cm\;34.3\times10^4\;Cells/cm^2$$ $$45\;cm\;27.2\times10^4\;Cells/cm^2$$ $$85\;cm\;26.3\times10^4\;Cells/cm^2$$ At temperatures of from 13.0 to $14.3^{\circ}C$, the distance travelled by the larvae (3.0 mm In shell length) averaged 11.36 mm for a Period of 30 days. Their locomation was relatively active between 6 p.m. and 9 p.m., and $52.2\%$ of them moved during this period. When the larvae (2.0 mm in shell length) were kept in water at $0\;to\;\~1.8^{\circ}C$, they moved 1.15cm between 4 p.m. and 8 p.m. and 0.10 cm between midnight and 8 a.m. The relationships between shell length and body weight of the abalone sampled from three different localities are shown as follows: Dolsan-do $W=0.2479\;L^{2.5721}$ Huksan-do $W=0.1001\;L^{3.1021}$ Pohang $W=0.9632\;L^{2.0611}$

• The Korean Journal of Malacology
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• v.28 no.1
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• pp.7-12
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• 2012

The effect of water temperature on spawning induction, larval development, spat settling and its growth of the cockle shell, Fulvia mutica, were investigated to obtain the basic data for effective seed production. The eggs, which were randomly divided into 6 groups of water temperatures of 14, 17, 20, 23, 26 and $29^{\circ}C$, were transferred into 1 L beaker, respectively. The relationships between the water temperature and the required time (1/h, hour) by each egg developmental stage were calculated. Biological minimum water temperature and the cumulative water temperature until egg development of the veliger stage were calculated to be $0.1^{\circ}C$ and $397.3^{\circ}C$, respectively. The optimal water temperature for developmental bioassay of F. mutica was clarified to be $23^{\circ}C$. The required time for the embryo to become veliger larvae was 20 hours at $23^{\circ}C$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.6 no.1_2
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• pp.76-80
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• 1973

Cyclina sinensis is an edible bivalve inhabiting wide tidal flats which are exposed to the air at ebb tide along the western coast of Korea. Over the period of June to September 1971, some specimens from a tidal flat near Inchon were submerged in sea water with various concentrations of ammonium hydroxide added and careful observations were made on their fertilization, early development, and metamorphosis of the larvae. The highest rate of fertilization was demonstrated by individuals treated with 1/1000 normal solution of ammonium hydroxide and their fertilized eggs followed normal development, i.e., two cell stage 1.5 hours after fertilization, blastular stage after 4 hours, and trochophore stage after 6 hours. Within 24 hrs after fertilization C. sinensis larvae have acquired the form of early straight-hinge veliger with the mean prodissoconch I with the length of $110\mu$. It takes seven days to get the umbo stage with the mean shell length of $190\mu$ and twenty days to get the morphosing stage with the mean shell length of $260\mu$. The larvae were cultured to the metamorphosing stage with the shell length of $270\mu$ in the laboratory condition.