• Journal of Marine Life Science
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• v.8 no.1
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• pp.32-42
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• 2023

Fish reproduction is regulated by various neurohormones secreted from the brain and gonadotropic hormones secreted from the pituitary. Reproduction of eel (Anguilla japonica) is also regulated by these hormones. However, how the neurohormones regulate the secretion of pituitary hormones during sexual maturation is not completely understood. Previous studies have shown that neurohormones such as progesterone (P4), melatonin and serotonin (5-HT) are involved in the regulation of reproductive processes in some fish. In this study, the eel pituitary was primary cultured, and stabilized pituitary cells were treated with P4, 17β-estradiol (E2), melatonin, or 5-HT. The effect of these treatments on the expression of FSHβ, LHβ, GH and SL mRNA was, then, investigated. P4 increased the expression of FSHβ and LHβ in pituitary cells, and melatonin increased the expression of GH and SL as well as FSHβ and LHβ. However, 5-HT did not significantly affect the expression of these mRNA. These results suggest that P4 and melatonin may play some important roles in the early sexual maturation of eels.

• Journal of Practical Agriculture & Fisheries Research
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• v.22 no.2
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• pp.51-58
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• 2020

Continuous mortality in commercially cultured Japanese eel (Anguilla japonica), showing symptoms of dermal ulcerations and focal hemorrhages on the body, occurred on a private farm in November, 2019 in Korea. A series of mortality had been described in one local eel culture farm from November to December in 2019. From the three cases, three isolates of Vibrio spp. were recovered from the blood, ascitic fluid, and kidney of the dead fish, respectively. Based on the 16S rRNA sequence comparisons, the Vibrio isolates from the 1^st and 3^rd cases (strain named 1E1-2 and 3K1-2) were identified as V. fluvialis and the isolate from the 2^nd case was identified as V. plantisponsor (strain named 2A3-1). Moreover, the 16S rRNA-based phylogenetic analysis revealed that strain 1E1-2 and 3K1-2 were most similar to V. fluvialis NBRC 103150^T, and strain 2A3-1 was most similar to V. plantisponsor NBRC103148^T. According to the results of the antibiotic resistance determination, V. fluvialis 1E1-2 showed intermediate resistance to tetracycline and chloramphenicol, and was resistant to trimethoprim-sulfamethoxazole. V. plantisponsor 2A3-1 showed intermediate resistance to ciprofloxacin and levofloxacin, and was resistant to trimethoprim-sulfamethoxazole. V. fluvialis 3K1-2 showed intermediate resistance to tetracycline, and was resistant to ampicillin and trimethoprim-sulfamethoxazole. These results have provided the evidences on the occurrence of antibiotic-resistant Vibrio infection in commercially cultured Japanese eels are present in Korea.

• Development and Reproduction
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• v.28 no.3
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• pp.75-86
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• 2024

Artificial sexual maturation of eel (Anguilla japonica) involves rearing in seawater and injecting salmon pituitary extract (SPE). The salinity of seawater and components of SPE influence hormonal activities of the eel pituitary, leading to gonad development. This study investigated the direct effects of salinity change and SPE treatment on the eel pituitary gland using primary cell cultures. Pituitary cells were cultured into four experimental groups: control culture (control), SPE-treated culture (SPE), NaCl-treated culture (NaCl) and NaCl+SPE NaCl+SPE treated culture (NaCl+SPE). We investigated the expression of genes presumably related to reproduction and/or salinity, including luteinizing hormone (LHβ), follicle stimulating hormone (FSHβ), progesterone receptor-like (pgrl), prolactin (PRL), dopamine receptor D4 (drd4), neuropeptide B/W receptor 2 (NPBWR2) and relaxin family peptide receptor 3-2b (rxfp3-2b). Gene expression analysis revealed significant upregulation of LHβ in SPE and NaCl+SPENaCl+SPE groups compared to control and NaCl (p<0.05). FSHβ expression did not show any significant changes. PRL showed a significant decrease in the NaCl group (p<0.05). Pgrl, NPBWR2, drd4, and rxfp3-2b displayed the highest expression in the control group, with downregulation observed in all treatment groups (NaCl, SPE, and NaCl+SPE) (p<0.05). This study demonstrated the direct effects of salinity changes and SPE treatment on the eel pituitary. Results from this study also suggest that salinity change is necessary but work together with SPE to induce reproductive process, and that LHβ, pgrl, PRL, drd4, NPBWR2, and rxfp3-2b genes are obviously associated with reproduction and salinity changes in eels.

• Journal of fish pathology
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• v.1 no.1
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• pp.5-30
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• 1988

This is a comprehensive study for considering the effective treatment and control program of bacterial disease occurring in common carp, israel carp, color carp, crucian carp, eel and tilapia by clarifying the causes, mechanism of infection and onset and the diagnostic criteria. As a first step, the authors investigated the external views, gross and histopathologic findings of diseased fish using 450 infected fishes obtained from various farmer of Korea. This infection was characterized by hyperemia, hemorrhage and swelling of body surface and fins, congestion of liver, spleen, kidney, inflammation of intestine, hemorrhagic inflammation of various tissues, and necrosis and ulcer of various tissues were accompanied in serious cases. Bacteriologically, Aeromonas hydrophila and Edwardsiella tarda were isoiated from these fishes. Particularly in the regular check on 222 eels, 177 strains were isolated as 29.94% of Aeromonas hydrophila, 48.58% of Edwardsiella tarda and 21.47% of Flexibacter columnaris. Hexibacter columnaris was isolated from corroded gill of eels. The identical disease was occurred by innoculating the isolated Aeromonas hydrophila and Edwardsiella tarda and the identical strains were isolated from infected experimental fishes. The eels which were diagnosed Aeromonas disease from Kwangju, Pusan accompanied hemorrhage, swelling of body surface and fins, inflammation of stomach and intestine containing mucous fluids mixed with the pathogens. Color carp and crucian carp which were innoculated with the isolated 5 strins of Aeromomas hydrorphil died within 3 or 4 days accompanying with the characteristics of Aeromonas disease. Edward disease was characterized by abscesses of body surface, pus formation with concentration on phagocytes. The size of absecsses increased with progression elf disease. There were also various abscesses at internal organ and white nodules appeared in kidney. Histologically, various progressive granuloma were examined without inflammation of intestine. Columnaris disease of eels showed no hemorrhage except slight white body color. In autopsy, most of internal organs appeared normal and there were no septic odors. The only character was corrosion of gills. In order to treat these bacterial diseases, infected fishes must bathe in 20ppm chloramphenicol or kanamycin solution for 1 hour. Besides, medication program in oral ingestion of 75mg/kg chloramphenicol per day continuing for 5 to 7 days. After injecting the formalin treated Aermonas hydrophila antigen into carp, relatively high agglutination titer showed between 3 weeks and 6 weeks. Though this titer decreased from that time, it was continued for 18 weeks. In the case of injecting the formalin treated Edwardsiella tarda antigen into tilapia, the titer also increased. But tilapia which were immersed in the suspension fluid of the formalin treated Edwardsiella tarda showed no increase of the titer.

• Journal of fish pathology
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• v.24 no.3
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• pp.237-245
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• 2011

Disease survey was conducted to investigate the cause of high mortality in 23 farms of eel, Anguilla japonica, during the period from 2000 to 2010. Seven kinds of fish pathogens were confirmed in the tested fish, which included: Pseudodactylogyrus sp. (infection rate: 65.5%, 19/29 samples), aquabirnavirus (ABV, 62.5%, 15/24 samples), Edwardsiella sp. (44%, 11/25 samples), anguillid herpesvirus 1 (AngHV1, 16.7%, 2/12 samples), Heterosporis sp. (10.3%, 3/29 samples), Vibrio sp. (8%, 2/25 samples), Trichodina sp. (3.4%, 1/29 samples). The rate of single infection was 44.8% (13/29 samples), while 62.1% (18/29 samples) showed mixed type of infection with 2 to 5 different pathogens. The most predominant mixed infection were Pseudodactylogyrus sp.-Edwardsiella sp. (10.3%, 3/29 samples), Pseudodactylogyrus sp.-ABV (10.3%, 3/29 samples), Edwardsiella sp.-ABV (6.9%, 2/29 samples). Infection by the above infectious parasites, bacteria and virus was confirmed in tested eels of juvenile to adult sizes that were cultured at $25{\sim}30^{\circ}C$ in almost the examined farms.

• Korean Journal of Environmental Biology
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• v.32 no.4
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• pp.382-388
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• 2014

Shortfin eel (Anguilla bicolor pacifica) is a species of commercial importance and its production is greatly affected due to the infection by Heterosporis anguillarum. In this study, we evaluated the effect of H. anguillarum infection on the growth of Shortfin eel. A disease that trunk muscle of cultured shortfin eel, Anguilla bicolor pacifica, were irregular and resulted in death, breakout of the commercial eel culture farm. We observed that the trunk muscle of infected eels were irregular and represented white or yellowish externally. Histopathologically, a great numbers of large or small spores and sporophorocysts were also observed in degenerated muscle layer. The cloning of specific gene of H. anguillarum, encoding small subunit ribosomal RNA (SSU-rRNA) was amplified by the polymerase chain reaction(PCR) from the muscle lesion of diseased eel. The size of clone gene is well matched with the size of small subunit ribosomal RNA of H. anguillarum and thus confirming the infection by H. anguillarum.

• Journal of fish pathology
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• v.25 no.3
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• pp.151-158
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• 2012

Diseased eel (Anguilla bicolor) displayed severe hemorrhages in the gills, and congestion and swelling in the liver. During the epizootic, the water temperature was $28^{\circ}C$ and the morality rates were about 5%. No parasites were found on the gills and skin. Bacteria were not cultured from any internal organs using TSA or SS agar at $28^{\circ}C$ for 48 hrs. Histopathologically, the gills showed epithelial hyperplasia in the base of secondary gill lamellae and hemorrhages in the capillaries. Some cells in the proliferated interlamellar epithelia exhibited marginal hyperchromatosis. And severe vacuolated changes in the parenchymal cells and congestion in the central veins were observed in the liver. The specific amplicon (396 bp) was detected from gills and opercula of affected eel PCR using Anguillid herpesvirus-1 (AngHV-1) -specific primer sets HVAPOLVPSD (5-'GTG TCG GGC TTT GTG GTG C-3') and HVAPOLOOSN (5'-CAT GCC GGG AGT CTT TTT GAT-3'). Sequencing analysis of the amplicon demonstrated that this gene was 99% homologous to the AngHV-1 sequence deposited in GenBank. This is the first report of AngHV-1 outbreak in the farmed shortfin eels (A. bicolor) in Korea. When diseased fish were maintained for 10 days at water temperatures of $32^{\circ}C$ and $35^{\circ}C$, the cumulative mortalities were 100% and 10%, respectively. Even though the AngHV-1 genome in the gills from the eel kept at $35^{\circ}C$ was detected using PCR, the structure of gill filaments was similar with that of normal fish. Increasing the water temperature to $35^{\circ}C$ was an effective way to diminish the mortality of AngHV-1 affected eel.

• Journal of Aquaculture
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• v.20 no.4
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• pp.219-225
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• 2007

This study investigated the correlationship between artificial maturation season and reproduction coefficient of cultured eel Anguilla japonica from May (spring) to next January (winter). The brood stock, female eels ($400{\sim}600\;g$) were artificially matured by weekly intramuscular injections of salmon pituitary extracts (SPE, 20 mg/fish) to induce a completion of vitellogenesis. After completion of vitellogenesis, final oocyte maturation and ovulation was induced by injection of $17{\alpha}$, $20{\beta}-dihydroxyprogesterone$ (DHP) at about $2\;{\mu}g/g$ body weight. Most fish ovulated $15{\sim}18\;h$ following the DHP injection. The ovulated fish were induced to natural spawning or artificial fertilization by the dry method. Males ($200{\sim}350\;g$) were received weekly intramuscular injections of human chorionic gonadotropin (HCG) at a dosage of 1 IU/g body weight to induce testicular maturation and spermiation. Seasonal reproduction coefficient which includes the rate of ovulation, buoyancy, fertilization and hatching of eggs in the artificially matured eel during spring to summer ($May{\sim}July$) were significantly higher than the other season, while there were no significant difference among spring and summer (P<0.05). Furthermore, the number of eggs spawned and larvae hatched in the artificially maturated eel during spring to summer ($May{\sim}July$) were significantly higher than the other season, while there were no significant difference in spring and summer (P<0.05). These results indicate that artificial maturation by hormone treatment of A. japonica was successful only during spring to summer, which is the maturation period in the wild stock in nature. Consequently, it is possible to determine the period of artificially induced sexual maturity by the reproduction coefficient which includes the rate of ovulation, buoyancy, fertilization and hatching of eggs in the cultured eel A. japonica.

• Journal of Fisheries and Marine Sciences Education
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• v.27 no.3
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• pp.672-681
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• 2015

Edwardsiellosis has become a serious diseases problem in cultured eels for many years. This study was performed to investigate possibility of vaccination against edwardsiellosis caused by Edwardsiella tarda. We conducted a immersion and/or oral vaccination using formalin-killed E. tarda in eel Anguilla japonica. Three groups of fish ($26.8{\pm}1.2g$, $7.1{\pm}0.7g$ and $2.2{\pm}0.4g$) were used in this study. The protection (relative percentage survival, RPS) and serum antibody response (agglutination titer) were evaluated in the vaccinated fish. No correlation between agglutination titer and survival rate was observed in vaccinated fish. However, there was a satisfactory protective (RPS>50%) in vaccinated fish. Immersion (10 mg/mL, 1 hr) and immersion (10 mg/mL, 1 hr) plus oral (10 mg/g, 10 days) of $26.8{\pm}1.2g$, immersion (10 mg/mL, 1 hr) plus oral (10 mg/g, 10 days) of $7.1{\pm}0.7g$ showed RPS of 62.6%, 52.2% and 56.8%, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.2
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• pp.282-290
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• 1997

Hepatocytes of Anguilla japonica have been prepared using a collagenase perfusion technique. The isolated cells attached efficiently to fibronectin-coated culture dishes and subsequently formed monolayers in serum-free medium. These cultures maintained in appropriate medium at least for 10 days with minimal cell loss. The effects of estradiol and pituitary hormones on vitellogenin (Vg) synthesis were examined in primary hepatocyte culture of the immature eels. In fish, as in other oviparous vertebrates, estrogen is a major inducer of Vg synthesis. However, $estradiol-17\beta(E_2)$ alone was insufficient to induce Vg synthesis in cultures of eel hepatocytes. Combination of $E_2$ with growth hormone (GH) and/or prolactin (PRL) markedly stimulated Vg synthesis. Even in cultures exposed to $E_2$ or precultured without hormones for 8 days, $E_2$ alone could not fully induce Vg synthesis. The synthesis of Vg was dramatically increased when hepatocytes were cultured in medium supplemented with $E_{2}+GH+PRL$ for 6 days. At this point, even though GH and/or PRL were eliminated from the medium, Vg synthesis was not influenced by these factors during culture of further 3 days. These results indicate that pituitary hormones, in particular GH and PRL, play important roles in the regulation of Vg synthesis in primary cultures of eel hepatocytes.