• Ocean and Polar Research
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• v.32 no.3
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• pp.247-254
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• 2010

Single cell PCR analysis and light and scanning electron microscopic techniques were utilized to identify free living bivalve larvae in the coastal waters of Tae-an, on the west coast of Korea. Through DNA sequencing, venerid clam larvae were isolated and identified as Ruditapes philippinarum (99% similarity) and Meretrix lusoria (99%). Under microscopic observation, the D-veliger stage of R. philippinarum exhibited symmetrical shoulder angles and an elliptical ventral form. In contrast, M. lusoria displayed asymmetrical shoulder angles and a round ventral form in the umbonal stage. Size of the R. philippinarum larvae was $156{\pm}22{\mu}m$ in length, $126{\pm}12{\mu}m$ in height, $92{\pm}14{\mu}m$ in width with a length: height ratio of 1.23. Meretrix lusoria was $202{\pm}44{\mu}m$ in length, $161{\pm}35{\mu}m$ in height, $96{\pm}38{\mu}m$ in width with a length: height ratio of 1.25. Experimental results indicate that morphological and molecular characteristics provide evidence for the larval identification of these two venerid clam larvae species in nature.

• Journal of Practical Agriculture & Fisheries Research
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• v.13 no.1
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• pp.131-136
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• 2011

For the establishment of seed production technique of warm water abalone species Haliotis sieboldii, development of the fertilized eggs and its biological minimum temperature were determined. The durations of each development stages at the six rearing temperature regimes were expressed as an exponential equation: 4 celled stage 1/h = 0.1346T - 2.1709(r² = 0.88) Morula stage 1/h = 0.0176T - 0.2184 (r² = 0.89) Trochophore 1/h = 0.0063T - 0.0512 (r² = 0.98) Veliger 1/h = 0.0045T - 0.0295 (r² = 0.99) 2nd c.t. 1/h = 0.0008T - 0.0047 (r² = 0.99) According to the equation, the biological minimum temperature for Haliotis sieboldii was estimated to be 9.7 ℃.

• The Korean Journal of Malacology
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• v.23 no.2
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• pp.189-198
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• 2007

The reproductive cycle, egg capsules in the egg-mass, first sexual maturity, and sex ratio of the bladder moon, Glossaulax didyma ($R\ddot{o}ding$) were investigated. The gastropods collected from the intertidal zone of Biin Bay, Seocheon, Korea were studied by using histological analysis and morphometric data. The gonadosomatic index (GSI) of females and males began to increase in March and reached maximum in May. Then their values sharply decreased from late in May to August due to spawning. The condition index (CI) began to increase in February and reached maximum in May, then gradually declined in the spawning period. The CI calculated for determination of the spawning period was coincided with changes in the GSI and gonadal phases. Spawning occurred between late in May to August in females and early in May to August in males. Spawning peak was observed between July and August when the seawater temperature rose to 19 $^{\circ}C$. Reproductive cycle with the gonadal development phases of this species can be divided into five successive stages in females and four in males: in females, early active stage (December to February), late active stage (February to March), ripe stage (April recovery stage (August to November); in males, active stage (December to March), ripe stage (March to July), copulation stage (early May to August), and recovery stage (August to January). Fully matured oocytes were approximately 250-270 ${\mu}m$ in size. The egg-mass was a hat in shape, and a number of egg capsules were found in an egg-mass. An egg capsule was 0.53-0.57 mm in size. An embryo (veliger larva) hatched from an egg capsule. Percentage of first sexual maturity in females and males were over 50% for individuals that are 40.1-45.0 mm in shell radius, and 100% for those that are over 45.1 mm. The sex ratio of female to male was significantly different from 1:1 $(x^2\;=\;57.22,\;p\;<\;0.05)$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.8 no.3
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• pp.185-195
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• 1975

The comparative studies Were conducted with respect to the artificial spawning early embryonic development, metamorphosis and growth of two species Meretrix lusoria and Cyclina sinensis collected from Inchon, Anmyon island and Buan areas from 1969 to 1974. The highest rate of artificial spawning of M. lusoria, which treated with a dilute ammoniun hydroxide(4/100-5/100N)-seawater solutions, was $25.0-33.3\%$, whereas in C. sinensis the rate of spawning was lower than that of M, lusoria under the similar experimental conditions$(12.5-19.0\%)$. However, the rate of artificial spawning of C. sinensis increased $40\%$ by repeated thermal stimulation. The rate of artificial fertilization of M. lusoria and C. sinensis showed highest value from those individuals which were treated with 1/1000N $NH_4OH$ solution. Their fertilized eggs, then, showed a normal development in the 1/1000N $NH_4OH$ solution. In the early embryonic development of M. lusoria and C. sinensis, the appearance of each of polar body, trochophore and D-shaped veliger were observed around 50min. 5-6 hours, and 23 hours after artificial fertilization respectively. The larval shell lengths of M. lusoria reached to $109,5{\pm}0.7\mu,\;144.6{\pm}1.3\mu$ and $208.0{\pm}0.0\mu$ around, 1, 11 and 20 days, after fertilization respectively. The larval shell lengths of C. sinensis reached to $110.5{\pm}0.6\mu,\;147.8{\pm}1.7\mu,\;and\;235.0{\pm}0.0\mu$ around 1, 10 ana 20 days, after fertilization respectively. The correlations of relative growth rate between the shell length(L) and sell height(H) found by the following simple formula from D-shaped veliger to metamorphosing stage. H=0.77L+6.82 for M. lusoria H=0.75L+8.50 for C. sinensis.

• The Korean Journal of Malacology
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• v.17 no.1
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• pp.45-55
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• 2001

Gametogenes, reproductive cycle, first sexual maturity(biological minimum size), sex ratio and larval development of the marsh clam Corbicula japonica were investigated monthly by histological observations. Samples were collected in brackish water of Gokgang stream, Kyungsangbuk-Do, Korea, from August 1997 to July 1998. Sexuality of Corbicula japonica is dioecious and the species are an oviparous clam. The gonads are irregularly arranged from the sub-region of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary is composed of a number of ovarian sac which are branched arborescent. Oogonia actively proliferate along the germinal epithelium of ovarian sac, in which young oocytes are growing. The testis is composed of a number of testicular tubules, and the epithelium of the tubule has function of germinal epithelium, along which spermatogonia actively proliferate. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed between developing oocytes and spermatocytes in the early developmental stages. With the further development of the ovary and testis these tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and eosinophilic granular cells are considered to be related to the growing of the oocytes and spermatocytes. The spawning period is from July to September, and the main spawning occur between July and August when seawater temperatures reach above 22$^{\circ}C$. The reproductive cycle of this species can be divided into five successive stages; early active (February to April), late active (May to July), ripe (June to September), partially spawned (July to September), degenerative (September to October) and resting stage (October to February). Percentages of first sexual maturity of female and male clams ranging in length from 10 mm to 12 mm are over 50% and 100% for clams over 16.0 mm in shell length. Fertilized eggs or Corbicula japonica were 80-90 ${\mu}{\textrm}{m}$ in diameter. In the early embryonic development of C. japonica, the appearance of polar body, trochophore and D-shaped veliger were observed around 40 min., 27 hours and 4 days after spawning, respectively, at a water temperature of 26.5-28.$0^{\circ}C$. The size of larvae of early umbo stage was about 185-210 ${\mu}{\textrm}{m}$ in shell length, 160-180 ${\mu}{\textrm}{m}$ in shell height around 7 days after fertilization. The correlation of relative growth between the culture day (D) and shell length (SL) was expressed by the following simple formula from D-shaped veliger to metamorphosing stage; SL = 13.300D + 209.36($r^2$= 0.9078).

• 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.21 no.1
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• pp.25-31
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• 2005

The embryos of marine bivalves have been commonly used in bioassays for the quality assessment of marine environments. Although several standard protocols for developmental bioassay with bivalves have been already proposed, there have been few trials for applying these protocols in environmental assessment, or for developing new protocol with Korean species. So, there is a strong need to establish the standard bioassay protocols using bivalves commonly found in Korean waters. Prior to developing a new protocol, it is essential to know the optimum conditions for the reliable bioassay procedures. Here, we established the purpose of this study to determine the optimum bioassay conditions for successful development of a common mussel, Mytilus galloprovincialis. The conditions considered as critical for developmental bioassay, and determined in this study were; (1) temperature, (2) salinity, and (3) initial density of embryo. The optimal temperature for developmental bioassay of M. galloprovincialis was determined as $15^{\circ}C$. At this temperature, the required time for the embryo to become veliger larva was 48 hr. The acceptable range of salinity for the embryotoxicity test using M. galloprivincialis was from 30 to 35 psu, which was narrower than that of the natural habitat of adult populations. The optimum density of embryo at the beginning of bioassay was 100 embryos/ml. Over this density, the proportion of normally developed larvae decreased significantly. The results obtained in this study will serve as a basis for preparation of the standard bioassay protocol using embryo of M. galloprovincialis.

• 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.

• Development and Reproduction
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• v.6 no.1
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• pp.37-44
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• 2002

First sexual maturity, sex ratio, spawning frequency, deposition of the egg capsules and fecundity of the female Rapana venosa(Valenciennes) inhabited in the artificially closed slag deposit area, Gwangyang Bay were investigated by histologicai and visual observations for natural living resource management. The rate of individuals reaching the first sexual maturity was 51.6％ in females measuring 7.1~8.0 cm in shell height, and 100％ in those > 10.1 cm. The total number of egg capsules per individual and the mean number of eggs in an egg capsule were 192~382 and 500, respectively. However, the number of eggs per individual and sizes of egg capsules under lower salinity and deficient food conditions in the closed slag deposit area were smaller than those under the optimum salinity and sufficient food conditions in the open regions. Fecundities of the species were approximately from 96,000 to 191,000 eggs/individual with two to low broods(spawning frequencies) during the spawning season. The duration of development in egg capsules was 18~19 days at about 18~2$0^{\circ}C$. R. venosa is a species whose embryos hatch as veliger larvae, not juvenile snail. The sex ratio of female : male was not significantly different from 1 : 1($\chi$$^2$= 0.23, p>0.05).

• 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.