• Journal of Marine Bioscience and Biotechnology
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• v.2 no.3
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• pp.142-148
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• 2007

An agar-degrading marine bacterium, strain AS-3 was isolated from the seawater. The strain AS-3 was identified as Algibacter lectus AS-3 by 16S rDNA sequence. The optimum medium for agarase activity of the isolated strain was determined to be marine medium, marine broth 2216 containing 0.1% agar as carbon source. An extracellular agarase was purified 6.9-fold from the culture supernatant by ammonium sulfate precipitation, ion exchange chromatography and gel filtration methods. The optimum pH and temperature for this enzyme were 7.0 and $40-50^{\circ}C$, respectively. Antioxidative activity of the strain AS-3 was 62.4% in the supernatant cultured for 12 h.

• Journal of Aquaculture
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• v.15 no.1
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• pp.7-13
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• 2002

Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.2
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• pp.115-118
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• 2000

The production of cultured coho salmon (Oncorhpchus kisutoh) in Korea has being increased year after year. Smolt being reared in freshwater suffer transferring into seawater and are farmed in cages for fattening. This handling processes including transportation, confinement into cages are unavoidable stress to fish in salmon farming and often end up to mass mortality, This study aimed to investigate the impact of handling process on the stress responses of coho salmon. The indicator of stress was measured by cortisol to be a first response, and for the second response test, glucose, triBlyceride, cholesterol, lactate and electrolyte of $K^+, Na^+, Cl^-$ in serum and the activities of alanine aminotrtnferase (ALT), aspartate aminotransferase(AST) and lactate dehydrogenase (LDH) were analyzed. As a result, the concentration of cortisol, glucose as well as LDH activity were significantly increased, whereas others showed no difference comparing with control group. It obviously demonstrated that handling process made fish stressful.

• Journal of Aquaculture
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• v.11 no.3
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• pp.285-294
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• 1998

When Cochlodinium polykrikoides came into the culture tanks through influent cultivated water during the red tides, hundred thousands of commercial flounders were concomitantly killed and many culturists suffered from a great deal of financial loss in the east coast of Korea. It is charactrized by high sinking rate after sunset and the formatino of clump which results in oxygen deficiency by its respiration at tank bottom under condition. We investigated the efficacy of hydrogen peroxide and chlorine dioxide, known to form radicals, for extermination of red tide organism C. polykrikoides. When C. polykrikoides seawater with a density of 6,000 cells/$m\ell$ was treated with 14, 28 and $42mg/\ell$ of hydrogen peroxide, its survival rate was markedly decreased to 9.8, 0.8 and 0.3% respectively immediately after 6 hours of treatments whereas when it was treated with 1.5, 2.1 and $3.0mg/\ell$ chlorine dioxide, its survival rate showed 87.7, 81.3 and 80.1 and 80.1% respectively at the same treatment time. Hydrogen peroxide was the effective agent since it has scarcely injured the cultured olive flounder when exposed to the tested concentration range of $14~28mg/\ell$ with the extermination of almost3 C. polykrikoides during the experimental period of 5 days and has shown the oxygen increase of approximately $1.23mg/\ell$ 2 hours immediately after the flounder by C. polykrikoides in the land-based culture tank is assumed to be not by the toxicity of itself but by oxygen dificiency from the rapid respiration of dinoflagellate clump sunken to the tank bottom.

• Korean Journal of Microbiology
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• v.41 no.1
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• pp.74-80
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• 2005

For the reseach of the natural marine antioxidant, an antioxidant-producing bacterium was isolated from seawater in Jeju costal area. The isolated strain SC2-1 was Gram-positive, catalase positive, oxidase negative, motile and small rods. The strain utilized sucrose, dextrose, fructose, mannitol and maltose as a sole carbon and energy source and NaCl required for growth. The radical scavenging activity of the culture supernatant was detemined by DPPH method. This bacterium was identified based on morphological, biochemical characteristics and 16S rDNA sequence analysis, and then was named Exiguobacterium sp. SC2-1. The optimum conditions of culture for production antioxidnat were $25^{\circ}C$, pH 6-8 and $4\%$ NaCl. The stain showed the highest activity and growth cultured in medium which added $1\%$ maltose. Hydroxyl radical activity of the supernatant of Exiguobacterium sp. SC2-1 was $73\%$. The SOD activity of the culture supernatant was estimated about $35\%$.

• Journal of Microbiology and Biotechnology
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• v.18 no.11
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• pp.1841-1847
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• 2008

Vibrio vulnificus is 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 the 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 1 g 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 fish pathology
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• v.12 no.1
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• pp.42-48
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• 1999

The disinfectant effects of total residual oxidants (TROs) produced by ozonization of natural sea water were investigated against fish pathogenic bacteria isolated from flounder and red seabream. The concentration of 0.1 mg TROs/liter was stable for 20 min in filtered natural seawater, and those of 0.3 and 0.5 mg TROs/liter were also stable for more 1 hr. Disinfectant effects of TRO against Edwardsiella tarda, Vibrio sp., Streptococcus sp. and Staphylococcus sp. were observed with a concentration of 0.1 mg/liter for 180 sec, and the treatment killed more than 99.9% of bacterial cells. With TROs of 0.3 to 0.5 mg/ liter, the viable cells of the bacteria were reduced by more than 99.99% in 60 sec.

• Fisheries and Aquatic Sciences
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• v.24 no.2
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• pp.89-98
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• 2021

The accumulation of ammonia, a consequence of intensive aquaculture activity, can damage cultured animals. We analyzed the survival rates, growth, antioxidant responses, and immune responses of abalones, Haliotis discus hannai (mean shell length 70.2 ± 4.9 mm; mean body weight 36.9 ± 3.6 g), biweekly for four weeks, to determine the accumulated concentrations of ammonia (0, 0.1, 0.2, 0.4, 0.8, and 1.6 mg/L). In our biweekly measurements, a survival rate of ≥ 95% was observed for all concentrations of ammonia. The specific growth rate (SGR) decreased at ≥ 0.8 mg/L (p < 0.05), but the daily increment in shell length (DISL) showed no significant change (p < 0.05). Regarding antioxidant responses, the superoxide dismutase (SOD) activity of the hepatopancreas was increased at ammonia concentrations over 0.8 and 0.2 mg/L (p < 0.05) at two and four weeks, respectively, and the SOD activity of the gills increased at concentrations over 0.4 and 1.6 mg/L (p < 0.05) at four weeks. The catalase (CAT) activity of the hepatopancreas and gills increased at ammonia concentrations > 0.8 mg/L (p < 0.05). Phenoloxidase (PO) activity increased at ammonia concentrations over 0.8 mg/L (p < 0.05), and Lysozyme (LZM) increased at concentrations over 0.8 and 0.4 mg/L (p < 0.05). Overall, our findings indicated that ammonia concentrations over 0.8 mg/L in seawater might damage the SGR, antioxidant responses, and immune responses in H. discus hannai. Our findings suggest the necessity of improvement or reinstallation for abalone aquaculture systems and can be used to assess the toxic effects of ammonia on H. hannai.

• Journal of fish pathology
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• v.37 no.1
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• pp.49-60
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• 2024

A viable but non-culturable (VBNC) state is a survival strategy adopted by bacteria when faced with unfavorable environmental conditions, rendering them unable to grow on nutrient agar while maintaining low metabolic activity. This study explored the impact of temperature and nutrient availability on inducing VBNC state in Edwardsiella piscicida, the most important bacterial fish pathogen, and assessed its pathogenicity at VBNC state. E. piscicida was suspended in filtered sterile seawater and exposed to three different temperatures (4, 10, and 25℃) to induce the VBNC state. Subsequently, the induced VBNC cells were subjected to resuscitation by either raising the temperature to 28℃ or inoculating them in brain heart infusion broth supplemented with 1% NaCl. A propidium monoazide (PMA)-qPCR method was also developed to selectively quantify live (VBNC or culturable) E. piscicida cells. The results showed that the bacteria entered the VBNC state after approximately 1 month at 4℃ and 25℃, and 2 months at 10℃. The VBNC E. piscicida cells were successfully revived within 3 days in a nutrient-rich environment at 28℃, highlighting the significance of temperature and nutrition in inducing and resuscitating the VBNC state. In pathogenicity tests, resuscitated E. piscicida cells exhibited high pathogenicity in olive flounder comparable to cultured bacteria, while VBNC cells showed no signs of infection, suggesting they are unlikely to resuscitate in fish. In conclusion, this study contributes to our understanding of fish pathogen ecology by investigating the characteristics of the VBNC state under varying temperature and nutrition conditions.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.29 no.1
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• pp.42-55
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• 2024

To discover unrecorded diatoms, seawater, sand, and pebbles were collected from the intertidal zone of five tidal flats from 2016 to 2023. Diatoms were isolated from the collected samples, cultured, and the ultrastructure of the cells was observed using a scanning electron microscope. 7 species of unrecorded diatoms, consisting of 3 orders, 5 families, and 7 genera, were discovered: Gomphoseptatum aestuarii, Rhoiconeis pagoensis, Seminavis exigua, Plagiolemma distortum, Staurotropis seychellensis, Biremis sigmoidea, Liriogramma sarcophagus. Among these, four genera, Rhoiconeis, Plagiolemma, Staurotropis, and Liriogramma, are reported for the first time in Korea. L. sarcophagus was separated from the central diatom Asteromphalus and transferred to Liriogramma, but its phylogenetic position has not yet been clearly established.