• 한국동물생명공학회지
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• 제35권3호
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• pp.223-231
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• 2020

Karyotype analysis is a major work in the process of triploid abalone production for the purpose of productivity and quality improvement. However, the metaphase spreads for karyotype analysis have been prepared just from the larvae at trochophore stage, which has restricted the spectrum of sample correction inhibiting more efficient analysis. Here, we investigated the feasibility of preparing metaphase spreads from the larvae at veliger stage that is the next developmental stage of trochophore. For this, diploid and triploid larvae at trochophore and veliger stages from Pacific abalone (Haliotis discus hannai) were subjected to metaphase spread preparation and its efficiencies were measured and compared each other. As the results, although the efficiencies of metaphase spread preparation were significantly lower in the larvae at veliger stage compared to the ones at trochophore stage regardless of ploidy status, we found that the preparation of metaphase spreads, which showed the clear chromosomal images containing the normal number of chromosomes, was possible from the veliger stage larvae. On the other hands, all larvae used in this study regardless of developmental stage and ploidy did not show colchicine sensitivity. Moreover, no significant difference was observed in cell cycle distribution of the cells comprising larvae between two developmental stages regardless of ploidy status. These suggested that the details of protocol to prepare metaphase spreads from abalone larvae should be optimized depending on its developmental stages. Taken together, we demonstrated the feasibility of preparing metaphase spreads from H. discus hannai veliger stage larvae for karyotype analysis.

• 한국발생생물학회지:발생과생식
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• 제17권4호
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• pp.337-343
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• 2013

Post-thawed larval rearing in Pacific oyster Crassostrea gigas was performed to investigate the survival rate with time course in three kinds of larvae cryopreserved. The highest survival rate and larval activity index (LAI) of post-thawed larvae were obtained from the permeation in 0.2 M sucrose and 2.0 M ethylene glycol (EG) at $-1^{\circ}C/min$ in freezing speed showing the survival rates just after thawing of 63.8% in trochophore, 84.1% in D-shaped veliger and 56.3% in early umbo veliger. In post-thawed larval rearing with food supply, the larvae lasted their lives until 24 hours in trochophore, 75 hours in D-shaped veliger and 57 hours in early umbo veliger. The results suggested that each larval stage post-thawed revealed no more further development to subsequent respective stage.

• 한국패류학회지
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• 제32권1호
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• pp.37-43
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• 2016

This study was carried out to determine the optimal water temperature for the embryonic development and laboratory culture of larvae of an intertidal mud snail, Nassarius festivus. The embryos and hatched veliger larvae of N. festivus were incubated at six different temperatures (5, 10, 15, 20, 25 and $30^{\circ}C$). Developmental time for each stage decreased as water temperature increased. The elapsed time to develop to the veliger larva at 15, 20, 25 and $30^{\circ}C$ was 559, 155, 131 and 103 hrs, respectively. At 5 and $10^{\circ}C$, embryo developed to veliger larvae but failed to hatch out of the egg capsule. In contrast, all embryos successfully hatched in the temperature range from 15 to $30^{\circ}C$. The biological minimum temperature during the embryonic development of N. festivus was estimated to be $9.5{\pm}0.4^{\circ}C$. The cumulative water temperatures for blastula, gastrula and veliger stages were calculated as $111{\pm}84$, $486{\pm}185$, $1,164{\pm}72^{\circ}C$, respectively. Temperature also affected the larval survival. Five days after hatching, more than 84% of larvae survived at all experimental temperatures. However, survival began to decrease after 6 days. It was 0% at $30^{\circ}C$. Survival of larvae incubated for 8 days was higher at 15 and $20^{\circ}C$ than other experimental temperatures. We therefore suggest that the optimal range of temperature for embryonic development and larval survival of N. festivus is $15-20^{\circ}C$.

• Environmental Engineering Research
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• 제22권1호
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• pp.108-115
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• 2017

Chlorination has been the most common antifouling method, but alternatives are under searching. In this article, we report how the hydrogen peroxide could enhance the effect of chlorination to prevent fouling by inhibiting larvae settlement and abatement of mussel colonization or by extinct of them; through marine mussel Mytilus edulis. The addition of hydrogen peroxide shows synergic effect on the veliger larvae (up to 19 folds) and effectively reduces required time of mussel mortality by 8-22%. For resolution of micro- and macro-fouling caused by the marine mussel, as well as diminishing of time and conventional chlorine dose could be important factor in favour of environment and economics.

• Ocean and Polar Research
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• 제37권1호
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• pp.37-47
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• 2015

The effects of low salinity (fertilization success and larval survival) on the limpet Cellana grata were studied at early stages of development using the marine bioassay technique. It was shown that, under normal conditions for development from fertilization to the post veliger stage, the salinity must be not less than 20.0~35.0 psu. However, the fertilization rate and larval survival of C. grata was obviously reduced at 5.0 psu and 10.0 psu, respectively. Mass mortality was estimated to occur at <20.0 psu (48-h $EC_{50}=19.54psu$) and the survival rate of normal veliger larvae decreased with experimental time during exposure. No observed effective concentration (NOEC) and lowest observed effect concentration (LOEC) of post veliger were estimated at 30.0 psu and 25.0 psu, respectively, during 48-h exposure. The tolerance limits of the test species to salinity revealed various concentration ranges of salinity, which may reflect the physiology and ecology of the initial development stages of C. grata. These results demonstrate that reduced salinity is detrimental to the reproductive success and larval survival of C. grata, and if salinity is lowered by natural or anthropogenic sources during spawning, this would lead to decreased reproductive success and larval settlement.

• Environmental Engineering Research
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• 제19권4호
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• pp.375-380
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• 2014

Macro fouling due to blue mussels (Mytilus edulis) has affected negatively on the operation efficiency and eventual system failure of offshore structures and coastal power stations. A certain range of chlorine (0.05, 0.1, 0.3, 0.5, 0.7 and 1.0 mg/L) was applied on the mussel larvae to identify the survival rate with respect to various exposure times under laboratory condition. The ciliary movement of the larvae was used to check their survival. The 1.0 mg/L of chlorine shows to 97% of larvae mortality whereas 0.7 mg/L of chlorine shows only 16% of larvae mortality. Minimum exposure times for 100% larvae mortality ranged from 300 to 20 min for increasing concentrations of chlorine (0.05~1.0 mg/L). It was found that 1 mg/L of chlorine was 4 times more efficient than 0.7 mg/L of that, and 15 times more than 0.05 mg/L of chlorine dose. Data collected and analyzed here will help plant operators to optimize chlorine dosage and its scheduling.

• 한국패류학회지
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• 제25권1호
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• pp.35-40
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• 2009

The trumpet shell Charonia sauliae is an endangered and valuable species with potential for aquaculture. For artificial propagation of C. sauliae, the effects of three different food microalgae on the development, growth, and survival rate of the larvae and spat were investigated. For the larval feeding experiments, we utilized six microalgae species as food sources, namely Pavlova lutheri, Tetraselmis suecica, Nannochloris oculata, Isochrysis galbana, Chaetoceros calcitrans, and Phaeodactylum tricornutum; for the larval and spat growth and survival experiments, we utilized T. suecica, C. calcitrans, and P. tricornutum. The results showed that the temporal digestion index (TDI) for the veliger larvae was significantly different for C. sauliae fed the different microalgae species (p < 0.05), that the T. suecica, C. calcitrans, and P. tricornutum cultivars were better suited for larval consumption (p < 0.05), and that the growth and survival of the larvae and spat were significantly influenced by food type, specifically P. tricornutum (p < 0.05). Further research is needed to evaluate the effects of other microalgae species, different algal concentrations, and biochemical composition on the growth and survival of C. sauliae.

• 한국발생생물학회지:발생과생식
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• 제18권1호
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• pp.25-31
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• 2014

This study was investigated spawning behavior, structure of egg masses and egg development in Aplysia kurodai inhabiting the coastal waters of Jeju Island, Korea. The mating and courtship behavior of A. kurodai occurred in the form of unilateral copulating with chain formation. In chain copulation, only the first animal acted as a female; the second and succeeding animals acted as males (sperm donors) to the animals in front and as females to the animals behind. The fertilized eggs were packaged in capsules that are embedded in jelly to form a cylindrical string called an egg masses. The number of capsule per cm of the egg masses was 55 to 60 capsules and each capsule within the egg masses held 15 to 25 eggs. After spawning, the egg masses were bright yellow or orange in color. This egg masses color not changed until embryos developed into trochophore stage. Thereafter, as embryo developed from trochophore stage to veliger stage the egg masses color became brownish. The fertilized eggs were spherical, with a diameter of approximately $80{\pm}1{\mu}m$ at spawning. At 5 to 6 days after spawning, the embryo developed into trochophore stage and began to rotate within the egg capsule. In the trochophore stage, the precursor of the velum, called the prototroch or prevelum, developed. At 10 days after spawning, the prevelum is transformed into the velum, and the trochophore developed into veliger stage. Between 10 to 15 days after spawning, the veligers broke out of the egg capsule, and hatched as free-swimming larvae.

• 한국패류학회지
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• 제19권1호
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• pp.33-40
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• 2003

To obtain the basic information on culture conditions for the larvae of Saxidomus purpuratus, experiments were conducted on the population from southern coast for (1) the success in fertilization and development from artificial fertilization among different months of a year, (2) the viability of sperms after exposure to seawater, (3) and the effects of temperature, salinity, and food organism on the survival and growth of larvae. Gametes obtained from dissection showed high rate of fertilization at all months. But the rate of development was higher only May-July. Developmental success seemed to be related with the quality of eggs at the time of fertilization. Developmental times for 2-cell, 4-cell, 8-cell, blastula, trochophore larva, and veliger larva at 20$^{\circ}C$ were 1.5, 2, 4, 18, 24, and 32 hr, respectively. Sperms could survive for more than 8 hr, however, actively swimming sperms could be found within 1 hr after exposure to seawater. It is recommended that sperms should be used for fertilization as soon as possible when they are exposed to seawater. At temperature of 35$^{\circ}C$, all the larvae died during 48 hr. Larval survival decreased when salinity was either lower than 20 psu or higher than 40 psu, and was 0% when salinity was 10 psu. Optimal range of temperature and salinity for rearing larvae of S. purpuratus were 20-25$^{\circ}C$ and 20-40 psu, respectively. Larvae grew from 111.5 to 235.3 ${\mu}$m during 21 days. Larvae fed mixed diets grew faster than unialgal diets. The fastest growth was observed when larvae were fed on the mixture of Isochrysis galbana and Nannochloris oculata.

• Ocean and Polar Research
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• 제32권3호
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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.