• Journal of Aquaculture
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• v.11 no.1
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• pp.11-17
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• 1998

Variations of hepatocyte in the larval liver of grouper, Epinephelus akaara wre examined to understand the effect of starvation during the first feeding period, 3 to 5 days after hatching. Total length of the fed larvae increased from the 5th day after hatching, although no significant difference between the fed and starved larvae was found untill the 4th day after hatching. Survival rate of the starved larvae decreased from the 4th day after hatching, and almost all of the larvae died by the 5th day after hatching. Nuclear size of hepatocyte in the starved larvae starterd to decrease from the 4th day after hatching. The sizes by 4th and 5th days after hatching in the starved larvae were 1.4 to 1.9 times smaller than those in the fed ones. Hepatocytes in the starved larvae showed irregular morphology in which the nuclei were irregularly shrunk and highly compacted from the 4th day, while hepatocyte nuclei in the fed ones maintained their uniform features during the whole experimental period. These results implied that the initial larval food should be supplied at least within the 4th day after hatching. Also, it suggested that the size of hepatocyte nucleus might be and indicator of starvation for wild and cultured grouper larvae.

• Journal of Aquaculture
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• v.11 no.2
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• pp.173-182
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• 1998

The study has been conducted to understand gonadal development and the effects of $17^{\alpha}$-methyltestosterone on sex inversion of the red spotted grouper, Epinephelus akaara. Fish were collected from Deukyand bay in the southern coast of Korea in August, 1996 and then they had been cultivated at the indoor tank until August, 1997. Gonad somatic index (GSI) in the females of both treated and control group began to increase from February when water temperature was rainse again, and reached the maximum value in August, whereas it had decreased from September adn thereafter maintained relatively low value until January. Unlike females, GSI in the male or intersex of treated groups decreased after June. Hepatosomatic index (HSI) of the control group tended to show the relatively low around Autumn, whereas it showed relatively highr value in April and June when the ovary was in the growing stage. Although the treated groups showed relatively higher value of the HSI than the control, hte paterns in monthly variation of HSI were similar to the control. Sexual change of the female grouper to the male was attempted by acceleration with oral administration of $17^{\alpha}$-methyltestosterone at the dose of 0.2 and 0.5mg/kg fish for 120days. Transitional hermaphroditic gonads were observed from the various size of groupers ranging 21.0 to 36.1 cm in total length, while the functional males could be induced from th individuals of 28.8 to 33.5cm in total length. This result indicated that larger groupers than 30cm in total length should be used for sex inversion to maleness with $17^{\alpha}$-methyltestosterone.

• Journal of Life Science
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• v.28 no.6
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• pp.639-647
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• 2018

A new heat shock protein 70 was identified in red-spotted grouper (Epinephelus akaara) based on an expression analysis. The cDNA of red-spotted grouper Hsp70 (designated RgHsp70) was cloned by the rapid amplification of cDNA ends (RACE) techniques. The full-length of RgHsp70 cDNA was 2,152 bp, consisting of a 5'-terminal untranslated region (UTR) of 105 bp, a 3'-terminal UTR of 274 bp, and an open reading frame (ORF) of 1,773 bp that encode a polypeptide of 590 amino acids with a theoretical molecular weight of 64.9 kDa and an estimated isoelectric point of 5.2. Multiple alignment and phylogenetic analyses revealed that the RgHsp70 gene shares a high similarity with other Hsp70 fish genes. RgHsp70 contained all three classical Hsp70 family signatures. The results indicated the RgHsp70 is a member of the heat shock protein 70 family. RgHsp70 mRNA was predominately expressed in the liver, with reduced expression noted in the head-kidney tissues. The expression analysis of different water temperatures (21, 18, 15 and $12^{\circ}C$) for sampled livers revealed that expression gradually increased at $12^{\circ}C$ compared to $21^{\circ}C$. In this study, the effects of water temperature lowering on the physiological conditions were investigated, and the results revealed that novel RgHsp70 may be an important molecule involved in stress responses.

• Journal of Aquaculture
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• v.11 no.4
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• pp.565-572
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• 1998

Survival rate of larval red spotted grouper, Epinephelus akaara is very low, because mouth size of the larvae is too small to eat normal sizes of rotifers. In order to enhance larval survival, optimal culture conditions for larvae using different live foods and water temperatures were studied. Bigger live food produced larger size of rotifer, when the rotifer was cultured with three different live foods such as Tetraselmis tetrathele, marine Chlorella and Nannochloris oculata. Also, the size of rotifer produced was temperature-dependent showing that higher water temperature induced smaller rotifer. Survial rate of the larvae increased with the increase of water temperature up to $29{\circ}C$. Also, higher larval survials were found at the food density of 15 to 20 individuals per milliliter. The larvae fed L-type or small individuals of L-type rotifer died all within a week, although the larvae fed S-type or small individuals of S-type one survived successfully during this period. The size of small individuals of S-type rotifer collected through fine net became larger in a few hours within larval culture tank. Survival rate of grouper larvae with initial stocking density of 40 thousands in 6-ton concrete tank was 6.3% in 10 days and 0.2% in 53 days after hatching, respectively. Total length of the larvae reached around 29.5 mm in 53days after hatching.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.4
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• pp.557-562
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• 2020

As part of an effort to establish of novel grouper hybrids for the aquaculture industry, we compared the embryonic development of fertilized eggs of giant grouper Epinephelus lanceolatus ♀×kelp grouper E. bruneus ♂ (GGKG) and giant grouper ♀×red-spotted grouper E. akaara ♂ (GGRG) with a maternal purebred (giant grouper ♀×♂, GG) at 27-29℃. The hatching rates were lower in GGKG (2.59%) and GGRG (0.36%) than in GG (12.29%). The hatching times were considerably longer in the two hybrids than in GG (22 h), which were similar between GGKG (28 h 30 min) and GGRG (28 h). The total lengths of yolk-absorbed larvae of GGKG (2.546±0.132 mm) and GGRG (2.602±0.093 mm) were similar to that of GG (2.565±0.149 mm). The deformity rates of hatched larvae (64.29-75.00%) did not differ between the two hybrids and GG. Although the hatching and deformity rates of GGKG and GGRG were inferior to those of GG, the possibility of mass production of the two hybrids larvae was demonstrated in this study.

• Journal of Aquaculture
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• v.10 no.4
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• pp.463-472
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• 1997

Floating and hatching rates, size and survival activity index(SAI) of spawned eggs of red spotted grouper, Epinephelus akaara were studied to understand egg qualities during the spawning periods in 1992, 1993 and 1995. The fish spawned from mid-July to mid-August when temperature ranged 23~$28^\circC$. Floating, fertilization and hatching rates were higher in the eggs from early to middle than those from late during the spawning period. SAI ranged 1.8~9.6, and it was closely related to survival rate until mouth operning time (SMOT). Egg diameter decreased gradually with the time of spawning, while diameter of oil globule was constant during the spawning periods. Induced spawnings were carried out with one injection of HCG over 5, 000 IU/kg body weight. Floating, fertilization and hatching rates of the induced spawning eggs were higher than those of untreated ones. Thus, induced spawning method with HCG was recommended for collecting fertilized eggs in bulk. Optimal time for the collection of high quality eggs was supposed to be from early to middle during the spawning periods. Considering that egg qualities were vulnerable within the period, hatchability, incidence of malformed larvae, SMOT and SAI were factors to be considered together to get good quality of eggs.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.5
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• pp.612-619
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• 2016

In this study, we crossbred Epinephelus akaara and E. lanceolatus to produce a hybrid grouper with faster growth and adaptation to domestic aquaculture environments. The plasma cortisol and glucose levels and osmoregulation (stress response indicators) of the hybrid grouper, E. akaara ♀ × E. lanceolatus ♂, were investigated under several salinity levels (32, 24, 16, and 8 psu). The body lengths and weights of E. akaara (8.2 ± 0.1 cm, 8.3 ± 0.4 g) and the hybrid (8.6 ± 0.1 cm, 10.0 ± 0.4 g) were similar at the start of the experiment, but were significantly different at the end of the experiment. Juveniles of both E. akaara and the hybrid showed greater weight gain, specific growth, and feed conversion rate (FCR) under low salinity of 16 psu. Under the 8 psu treatment, the juvenile E. akaara all died, while the hybrid juveniles survived. Plasma cortisol levels were not affected by lower salinity in both species. The above results indicate that the hybrid is more tolerant of low salinity than is E. akaara, although both species exhibited higher growth and FCR at 16 psu, lower than the salinity of natural seawater. Thus, juveniles of both E. akaara and the hybrid can be more effectively cultured in brackish areas or waters with salinity lower than that of seawater.

• Journal of Aquaculture
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• v.10 no.4
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• pp.473-483
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• 1997

Yolk resorotion and commencement of external feeding in larvae of red spotted grouper, Epinepheus akaara were studied to understand the high mortality of larvae. Consumption of yolk was higher than that of oil globule for the first 24hs after hatching. The resorption rates of yolk and oil globule increased with the increase of temperature. The time yolk was comsumed up to 99% was 84hs after hatching in 23~$25^{\circ}C$,72hs at ~$27^{\circ}C$, and 60hs in 29~~$31^{\circ}C$, while the time for oil globule was 96hs in 23~$25^{\circ}C$, 84hs at $27^{\circ}C$, and 60hs in 29~$31^{\circ}C$. The relationship between mouth openning time (Y) and water temperature (X) was repersented as a linear regression, Y=-2.53X+118.4, (r2=0.89). In 72~96hs after hatching, upper jaw sizes of the larvae reached around 0.114 mm. By the time the larvae comsumed most of the yolk and oil globule, and started external feeding. The relationship between upper jaw length (Y) and total length (X) was represented as a linear regression, Y=0.139X-0.165, (r2=0.88). Rotifer was not found in the guts of larvae before 72hs after hatching at $28^{\circ}C$, when larvae resorbed their yolk and oil globule over 99%. But, at 96hs after hatching, about 33% of the larvae had the rotifer in their guts. Delayed feeding experiments revealed that the earlier food supply within 84hs after hatching showed the better survival rates for the larvae. The larvae fed at 96hs after hatching died all within 120hs.

• Journal of Life Science
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• v.24 no.9
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• pp.995-1000
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• 2014

The studies of marine viruses in terms of viral isolation and detection have been limited due to the high mutation rate and genetic diversity of marine viruses. Of the modern methods currently used to detect marine viruses, serological methods based on enzyme-linked immunosorbent assay (ELISA) are the most common. They depend largely on the quality of the antibodies and on highly purified suitable antigens. Recently, a new experimental system for using viral capsid protein as an antigen has been developed using the yeast surface display (YSD) technique. In the present study, the capsid protein gene of the red-spotted grouper nervous necrosis virus (RGNNV) was expressed and purified via YSD and HA-tagging systems, respectively. Two regions of the RGNNV capsid protein gene, RGNNV1 and RGNNV2, were individually synthesized and subcloned into a yeast expression vector, pCTCON. The expressions of each RGNNV capsid protein in the Saccharomyces cerevisiae strain EBY100 were indirectly detected by flow cytometry with fluorescently labeled antibodies, while recognizing the C-terminal c-myc tags encoded by the display vector. The expressed RGNNV capsid proteins were isolated from the yeast surface through the cleavage of the disulfide bond between the Aga1 and Aga2 proteins after ${\beta}$-mercaptoethanol treatment, and they were directly detected by Western blot using anti-HA antibody. These results indicated that YSD and HA-tagging systems could be applicable to the expressions and purification of recombinant RGNNV capsid proteins.

• Journal of fish pathology
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• v.34 no.1
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• pp.99-104
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• 2021

In this study, we investigated safety and efficacy of a low temperature immunization protocol with NNV in red spotted grouper, Epinephelus akaara and long tooth grouper, Epinephelus bruneus and seven band grouper, Hyporthodus septemfasciatus. Further, growth rate between immunized and naïve fish was evaluated during the experiment to check side effect of immunization. Three grouper species were immunized by immersion method with live NNV at 10^5.0 TCID₅₀/mL at 16.5℃ for 30 min and reared for 120 days at natural sea water temperature. To evaluate growth rate, total length and wet weight was measured 7 times after immunization. Immunized three grouper species were challenged by intramuscular inoculation with NNV at 10^4.2 TCID₅₀/100 µL/fish. Immunization at low temperature with live NNV did not show any clinical symptoms of infection, mortality and inhibition of growth. After challenge, cumulative mortality of naïve seven band grouper, red spotted grouper, long tooth grouper were 45, 10, 20 %, respectively. However no mortality was observed at immunized groupers. Thus, it was demonstrated that immunization at low temperature with live NNV are able to protect three different species of groupers without inhibition of growth.