• Korean Journal of Fisheries and Aquatic Sciences
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• v.46 no.3
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• pp.252-258
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• 2013

The quality and quantity of food organisms in fish seed production are important. The marine microalgae Nannochloropsis oculata are used as initial food organisms in the field. We investigated the effects of salinity (0, 10, 20, 30, 40 and 50 psu) on the lipid and fatty acid composition of N. oculata. Cultivation of N. oculata at varying salinities showed the highest growth rate at 20 psu. Total lipid content ranged from 17.26 to 18.63% at salinities from 0 to 50 psu). The nonpolar lipid content increased markedly at 30 psu and was highest at 15.55%. The polar lipid content was lowest at 30 psu, by 84.45%. It was also found that the omega-3 and EPA contents were inversely proportional to salt concentration. For the polar and nonpolar lipid compositions, there was no significant effect of salinity. Omega-3 polyunsaturated fatty acid content especially the content of EPA in the seawater larvae is the essential fatty acid in this food organism. It is thus advantageous to culture N. oculata at 20 psu.

• Journal of Marine Bioscience and Biotechnology
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• v.2 no.4
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• pp.263-266
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• 2007

Vibrio vulnificusis a causative agent of serious diseases in humans resulting from the contact of wound with seawater or consumption of raw seafood. Several studies aimed at detecting V. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. In this study, we targeted the rpoS gene, a general stress regulator, to detect V. vulnificus. PCR specificity was identified by amplification of 8 V. vulnificus templates and by the loss of a PCR product with 36 non-V. vulnificus strains. The PCR assay had the 273-bp fragment and the sensitivity of 10 pg DNA from V. vulnificus. SYBR Green I-based real-time PCR assay targeting the rpoS gene showed a melting temperature of approximately $84^{\circ}C$ for V. vulnificus strains. The minimum level of detection by real-time PCR was 2 pg of purified genomic DNA, or $10^3$ V. vulnificus cells from pure cultured broth and $10^3$ cells in 1g of oyster tissue homogenates. These data indicate that real-time PCR is a sensitive, species-specific, and rapid method for detecting this bacterium using the rpoS gene in pure cultures and in infected oyster tissues.

• Journal of Microbiology
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• v.42 no.2
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• pp.74-79
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• 2004

Ingestion of shellfish-associated Vibrio parahaemolyticus is the primary cause of potentially severe gas-troenteritis in many countries. However, only Kanagawa phenomenon (hemolysin) positive (KP$\^$+/) strains of V. parahaemolyticus are isolated from patients, whereas >99％ of strains isolated from the environment do not produce this hemolysin (i.e. are KP$\^$-/). The reasons for these differences are not known. Following a temperature downshift, Vibrio parahaemolyticus enters the viable but noncultur-able (VBNC) state wherein cells maintain viability but cannot be cultured on routine microbiological media. We speculated that KP$\^$+/ and KP$\^$-/ strains may respond differently to the temperature and salinity conditions of seawater by entering into this state which might account for the low numbers of cul-turable KP$\^$+/ strains isolated from estuarine waters. The response of eleven KP$\^$+/ and KP$\^$-/ strains of V. parahaemolyticus following exposure to a nutrient and temperature downshift in different salinities, similar to conditions encountered in their environment, was examined. The strains included those from which the KP$\^$+/ genes had been selectively removed or added. Our results indicated that the ability to produce hemolysin did not affect entrance into the VBNC state. Further, VBNC cells of both biotypes could be restored to the culturable state following an overnight temperature upshift.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.5
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• pp.601-606
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• 1999

Physiological responses of cultured olive flounder (Paralichthys olivaceus) on lowering seawater temperature sharply and continuously were studied with 4 experiments of temperature changes (Exp.I$\~$IV). In Exp.1, the temperature was decreased from $18^{\circ}C$ to $9^{\circ}C$ by the rate of $1^{\circ}C$/hr, thereafter back to the initial temperature after 5 dars. With the same conditions of temperature rate and 5 days interval, the temperature changes for Exp.II, III and IV were $20^{\circ}C$ to $17^{\circ}C,\;23^{\circ}C$ to $14^{\circ}C$ and $23^{\circ}C$ to $17^{\circ}C$, respectively, Serum cortisol and glucose were measured during whole experiments. Hematocrit (Ht), hemoglobin (Hb), red blood cell (RBC) and mean corpuscular hemoglobin concentration (MCHC) were measured in the Exp.I, and osmolality, electrolytes ($Na^+,\;Cl^-,\;K^+,\;Ca^{2+}$), total protein, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) of serum, in Exp.II$\~$IV. Serum cortisol levels were significantly increased by the lowering temperature sharply during whole experiments, while serum glucose levels were increased only in Exp,III and IV. Ht, RBC and Hb were decreased as the water temperature was lowered, but MCHC was increased. The serum osmolality was reduced and the unstable changes of electrolytes were shown by the changes of seawater temperature. No significant changes in total protein, ALT and AST activity were observed.

• Journal of Marine Life Science
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• v.7 no.2
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• pp.171-179
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• 2022

The present study investigated the survival rate, respiration rate, plasma stress index, and histological changes according to exposure time of cultured red seabream (Pagrus major) and olive flounder (Paralichthys olivaceus) exposed to Cochlodinium polykrikoides red tide. Fish cultured in natural seawater were used as the control group. Cochlodinium polykrikoides density was set to 5,500±200 cells·ml^-1 in the experimental groups. All red seabreams died within 1 hour of exposure to red tide, whereas all olive flounders died within 5 hours of exposure. Analysis of physiological response revealed that in red seabream, plasma glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), catalase (CAT), and glutathione peroxidase (GPx) concentrations were increased; plasma glucose and superoxide dismutase (SOD) concentration were decreased. Meanwhile, in olive flounders, plasma cortisol, GOT, and GPT concentrations were increased; plasma glucose concentrations were increased during the first hour of exposure, followed by decrease after 5 hours; and plasma SOD, CAT, and GPx concentrations decreased during the first hour of exposure. Histological analysis revealed structural damage to the gills of both red seabream and olive flounder. In conclusion, the exposure of red seabream and olive flounder to Cochlodinium polykrikoides red tide at the density of 5,500 cells·ml^-1 induces oxidative stress, which activates antioxidant defense mechanisms, ultimately leading to liver and gill damage.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.6
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• pp.812-813
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• 1995

Lactococcal cells are nutritionally fastidious and thus, generally cultured either in milk or M17 medium (Terzaghi and Sandine, 1975). In this study, Lactococcus cremoris wild-type (KH) and its less­proteolytic mutant (KHA1) cells were grown on the M17 medium or with modified M17 medium by replicated parallel experiments. The modified M17 medium had the same composition as M17 medium, except that lactose was replaced by glucose. Analyses of culture-broth samples, in which the M17 and the modified M17 media were used, were conducted by high-performance liquid chromatography (HPLC). But, working with these media created noisy problems in analyses of samples. Therefore, a new semi-synthetic medium was developed on the basis of nutritional requirements (Morishita et al., 1981). The composition of the semi-synthetic medium determined on the basis of the nutritional requirements and the composition of milk, is presented in Table 1. The composition of M17 medium is also presented and compared in the table. L. cremoris KH and KHA1 cells were grown again on the new synthetic medium containing glucose or lactose. The broth samples were then drawn and analyzed by HPLC. Clearer separations of fermented products were achieved from the new medium than those with the M17 and the modified M17 media. In comparison with the M17 or the modified M17 media, growth on the new medium was good (Kim et al, 1993). Additional fermentations were also carried out at a controlled pH of 7.0, where enhanced growth of lactococcal cells was obtained. In the fermentations, samples were also analyzed for the concentrations of sugar and lactic acid. The results showed that the new synthetic medium was as good as or better than the M 17 and the modified M 17 media. This is because casein hydrolysate in the synthetic medium provided a ready supply of amino acids and peptides for L. cremoris KH and KHA1 cells. Lactic acid bacteria (LAB) including Lactococcal cells have been known to be an effective means of preserving foods, at the same time as giving particular tastes in fields of dairy products. LAB also have always occupied an important place in the technology of sea products, and marine LAB have known to be present in traditional fermented products (Ohhira et al, 1988). To apply the new synthetic medium to marine LAB, two different LAB were isolated from pickled anchovy and pollacks caviar and were grown on the new media in which various concentrations of NaCl $(3, 5, 7 and 10\%)$ added. They were also grown on the medium solution in natural seawater $(35\%o\;salinity)$ and on the solution of natural seawater itself, too. As seen in Fig. 1, Marine LAB were grown best on the synthetic medium solution in natural seawater and the higher concentrations of NaCl were added to the medium, the longer lag-phase of growth profile appeared. Marine LAB in natural seawater were not grown well. From these results, the synthetic medium seems good to cultivate cells which are essential to get salted fish aged. In this study, it showed that the new synthetic medium provided adequate nutrition for L. cremoris KH and KHA1 cells, which have been used as cheese starters (Stadhouders et al, 1988). Using this new medium, the acid production capability of starter cultures could be also measured quantitatively. Thus, this new medium was inferior to the M17 or the modified M17 medium in culturing the cheese starters and in measuring fermentation characteristics of the starter cells. Moreover, this new medium found to be good for selected and well-identified marine LAB which are used in rapid fermentations of low-salted fish.

• Journal of Aquaculture
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• v.19 no.1
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• pp.19-24
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• 2006

The objective of this study was to develop an efficient control method for the Alella macrotrachelus parasitic cultured black porgy, Acanthopagrus schlegeli utilizing the superior osmoregulatory ability of the host fish. The average number of the parasites on a fish was 5 worms (all female), which were attached to the cartilage of gill filament by their bulla. Morphologically, the parasite was tripartite with each length of $1822.1{\pm}521.5{\mu}m$ for the trunk $1825.0{\pm}495.8{\mu}m$ for the cephalothorax, and $134.2{\pm}43.1$ for the bulla. In histological observations, it was found that the female parasite took gill lamellae. Damage and loss of gill lamellae by the parasite caused hemorrhage and anemia in the fish. All parasites died within 48 hours by osmotic shock of freshwater or $5{\sim}15\%_{\circ}$ seawater treatment. These results suggest that Alella macrotrachelus could be controled by hyposmotic treatment.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.25 no.2
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• pp.26-41
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• 2020

In order to culture a life for the physiological and ecological research of the meiofauna, this study aimed to identify the most ideal condition in which the meiofauna can be cultured within a laboratory by setting various environmental conditions. The sediment deposits and seawater were collected from the intertidal zone in Mallipo of the west coast. A aquarium in which the internal environment can be controlled by constantly maintaining the temperature and humidity was fabricated and the culture experiments of the collected meiofauna were conducted together with the sea water and sediment deposits collected. The experiment 1 was conducted after establishing the similar environment as the collecting location. Under the same condition as the experiment 1, the experiment 2 verified a difference between when live foods were supplied and were not. In the experiment 3, the changes in the meiofauna colony were checked according to with or without light and live foods. In the results of culturing experiments, the habitat density and the number of appeared classification groups of the meiofauna colony were relatively higher both in the water tank with supplying the live foods and under the condition of having light in 12-hour cycle than those in the aquarium without live foods and under no light condition. In addition, the habitat density of meiofauna cultured within a laboratory exhibited relatively higher value than that under the natural state.

• Journal of fish pathology
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• v.19 no.1
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• pp.7-15
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• 2006

Amyloodinium sp. was found on the gills of mullet (Mugil cephalus) cultured on land. No external symptoms in the diseased fish were found except decoloration of the gills. In fresh preparations of the gills the parasites were opaque round or oval shape with a bright nucleus and 43.5 ㎛ (18.2～72.7, n=20) in size. In preparations added a drop of Lugol solution, they were black with the same shapes in fresh preparations and 43.5 ㎛ (n=20) in size. The parasites were stained black and blue in a droplet of Lugol solution and Diff-Quick III solution, respectively and their sizes were a little larger than in wet preparations. After stained with May-Grunwald Giemsa, the parasites appeared granular eosinophlic in the peripheral cytoplasm and granular strong basophilic in the center. In silver impregnated specimens the peripheral granules were negative and the central ones positive. The granules appeared brown in purplish cytoplasm after staining with Lugol solution. The parasites developed by binary division when they were cultivated in filtered seawater at 20℃. Histopathologically severe epithelial hyperplasia and fusion in the gill filaments resulted in clubbing, especially the proximal region of the filament. Epithelial hyperplasia was also found in the basal regions of the gill filaments and some epithelial cells were occasionally detached from the filaments. Some pear-shaped trophonts of the parasites with rhizoid attached on the gill filaments showing hyperplasia of the epithelial cells and mucous cells.

• Journal of Veterinary Clinics
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• v.26 no.3
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• pp.226-230
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• 2009

We evaluated the efficacy of $\beta$-hemolytic Streptococcus(S.) iniae vaccine on cultured olive flounder. Three hundred flounders(weight $50{\pm}5$ g) were obtained from two farm at Wando and Taean in the southern and western coast of Korea at May and June 2007, respectively. Twenty of flounders moved in 0.5 tons aquaria in land-marine tank system of National Veterinary Research and Quarantine Service. Seawater was transported from the sea of Inchon in western Korea, and water temperature maintained to $22^{\circ}C$ and $25^{\circ}C$ during the vaccination and challenge test, respectively. We used the formalin-inactivated $\beta$-hemolytic S. iniae vaccine produced by domestic manufacturers. The vaccine was intraperitoneally administered to fish. The vaccinated and control group were challenged with intraperitoneal injection by virulent S. iniae SI-36 isolates with $5.0{\times}10^8$ CFU/fish at 3 weeks after vaccination. We evaluated the vaccine efficacy by calculating numbers of dead fish, and observing of clinical signs, exterior and gross lesions, and examining bacteria isolation and identification. Thirty-four(25.2%) of 135 control and vaccinated group fish were dead with serious anemia, abdominal extension, and hernia of intestine during 3 weeks post vaccination. We isolated Neoheterobothrium hirame from the buccal cavity and Edwardsiella tarda from kidney of dead and diseased fish. When infected fish with these agents were challenged with S. iniae SI-36 isolates, the cumulative mortality of control and vaccinated group were 86.7, and 46.7%, respectively. However, significant differences(p<0.05) were observed on cumulative mortality between control(20.0%) and vaccinated group(95.0%) at second trials with 40 healthy, and relative percent survival(RPS) was 78.0%. We confirmed that the efficacy of $\beta$-hemolytic S. iniae vaccine on olive flounder were impacted by health condition such as bacterial and parasitic diseases.