• Korean Journal of Ichthyology
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• v.31 no.1
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• pp.23-29
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• 2019

The embryonic development and hatchability of the artificially fertilized eggs of convict grouper (Hyporthodus septemfasciatus) ♀${\times}$giant grouper (Epinephelus lanceolatus) ♂ (CGGG) were compared to those of the maternal pure species (convict grouper ♀${\times}$♂, CG) to establish a novel grouper hybrid for aquaculture industry. The fertilized eggs were divided into nine 5-L beakers (3,000~5,000 eggs/beaker) filled with UV sterilized seawater and incubated at a temperature range of $23.5{\sim}24.8^{\circ}C$ (32.1~32.8 psu). The percentages of fertilization and hatching of CGGG were $69.4{\pm}1.5%$ and $59.0{\pm}5.1%$, respectively and were significantly lower than those of CG (p<0.05). The CGGG proceeded normal embryonic development similar to that of CG, but showed an irregular cleavage, immature embryonic body and spinal deformity in hatched larvae. The incubation time from fertilization to hatching of CGGG was 31 hrs, which was approximately 2 hrs slower than that of CG. Our study provided the possibility of mass production of grouper hybrid CGGG larvae.

• Journal of fish pathology
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• v.37 no.1
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• pp.97-110
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• 2024

The Serranidae is high-quality fish species with good meat quality and is traded at high price, and is attracting attention in South Korea as a cultured species that creates high added value. However, the high-density fish farming for mass production increases the risk of mass mortality due to infectious diseases, leading to enormous economic losses. Therefore, in order to safely prevent and protect farmed fish from serious infectious diseases, it is necessary to conduct disease monitoring on a regular basis. In this study, Hyporthodus septemfasciatus, Epinephelus moara, and the hybrid longtooth grouper (E. moara ♀×E. lanceolatus ♂) were collected once a month from fish farm of Garorim and Aquabiotech Co., Ltd for a total of six months, from July to December 2023. We investigated infections of five species of bacterial diseases, including Flavobacterium columnare, six species of viral diseases, including LCDV (lymphocystis disease virus), and parasitic pathogens in grouper farms. As the result, Vibrio vulnificus and V. harveyi were detected in H. septemfasciatus in August, in the case of viral diseases, NNV was detected in H. septemfasciatus from July to August using RT-PCR or PCR. Finally, In the case of parasitic diseases, Tricodina sp. was detected in E. moara and the hybrid longtooth grouper from August to December.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.1
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• pp.31-37
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• 2021

The mass production of fertilized eggs of the convict grouper Hyporthodus septemfasciatus was studied from 2013 to 2020 for industrial aquaculture. The experiment was divided into two groups. Group 1 broodstock was raised from wild-caught fry and used from 2013 to 2020. Group 2 broodstock was raised from artificially propagated fry and used from 2019 to 2020. Males used to collect sperm for artificial insemination weighed more than 7 kg. The effects of various hormones on artificial ovulation were investigated from 2013 onward. Among these, luteinizing hormone-releasing hormone analogue (LHRHa) at 100 ㎍/kg body weight showed the most effective results and was used for artificial egg collection from 2014 onward. In Group 1, the average total egg production per year, average egg production per individual, fertilization rate, and hatching rate were 26,143 mL, 609.7 mL, 93.3%, and 91.8%, respectively, and in Group 2, were 2,750 mL, 316.5 mL, 92.1%, and 90.4%, respectively. Based on these results, we showed that a large number of fertilized eggs for artificial seeding could be produced consistently. Moreover, the mass production of fertilized eggs in Group 2 establishes a foundation for the complete aquaculture cycle of H. septemfasciatus.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.5
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• pp.527-533
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• 2017

We investigated the infection of nervous necrosis virus (NNV) in seven band grouper Hyporthodus septemfasciatus broodstocks, which have been reared in aquaculture farms in South Korea during 2012-2014. To investigate the prevalence of NNV within the broodstock, egg, sperm, and blood were sampled in the spawning season. The egg and sperm samples were subjected to a nested reverse transcription (RT) polymerase chain reaction (PCR) assay to detect NNV and were inoculated on SSN-1 cells to culture the virus. Blood samples were used to detect antibodies against NNV using enzyme linked immunosorbent assay (ELSIA). Positive values from ELISA were found in 39 of 162 samples (24%) in 2012, and 13 of 28 samples (46%) in 2014. Additionally, 4 of 34 broodstocks (11%) investigated in 2013-2014 were determined to be carriers from the nested RT-PCR and in vitro cultivation. The broodstocks in which antibodies against NNV were detected by ELISA, or in which NNV was detected by the nested RT-PCR assay, posed a risk of vertical transmission of NNV. Therefore, it is necessary to select virus-free broodstocks in seed production to reduce the possibility of the vertical transmission of NNV.

• Fisheries and Aquatic Sciences
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• v.22 no.12
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• pp.28.1-28.8
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• 2019

Background: In the present study, we evaluated four commonly used housekeeping genes, viz., actin-β, elongation factor-1α (EF1α), acidic ribosomal protein (ARP), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as internal references for quantitative analysis of immune genes in nervous necrosis virus (NNV)-infected seven-band grouper, Hyporthodus septemfasciatus. Methods: Expression profiles of the four genes were estimated in 12 tissues of healthy and infected seven-band grouper. Expression stability of the genes was calculated using the delta Ct method, BestKeeper, NormFinder, and geNorm algorithms. Consensus ranking was performed using RefFinder, and statistical analysis was done using GraphpadPrism 5.0. Results: Tissue-specific variations were observed in the four tested housekeeping genes of healthy and NNV-infected seven-band grouper. Fold change calculation for interferon-1 and Mx expression using the four housekeeping genes as internal references presented varied profiles for each tissue. EF1α and actin-β was the most stable expressed gene in tissues of healthy and NNV-infected seven-band grouper, respectively. Consensus ranking using RefFinder suggested EF1α as the least variable and highly stable gene in the healthy and infected animals. Conclusions: These results suggest that EF1α can be a fairly better internal reference in comparison to other tested genes in this study during the NNV infection process. This forms the pilot study on the validation of reference genes in Hyporthodus septemfasciatus, in the context of NNV infection.

• 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.

• Journal of fish pathology
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• v.35 no.1
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• pp.121-128
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• 2022

Recent studies revealed that histone proteins are involved in innate immune responses during pathogen invasion as well as DNA packing. This study characterized the histone genes (H2A.V) of sevenband groupers and analyzed gene expression in NNV-infected sevenband groupers. The open reading frame (ORF) of H2A.V is 387 bp which encoded 128 amino acid residues. The deduced amino acid sequence of H2A.V harbor a highly conserved domain for H2A/H2B/H3 and H2A_C binding domain. Quantitative real-time PCR analysis showed that H2A.V had a high gene expression level in the brain and blood after being NNV-infected. An increase in extracellular histone protein in the blood has been identified as a biomarker for vascular function in humans. More research is required to understand histone's immune response at the protein level or in aquatic animals.