• Korean Journal of Fisheries and Aquatic Sciences
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• v.52 no.4
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• pp.382-388
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• 2019

Streptococcosis caused by Streptococcus parauberis is a very important disease in farmed olive flounder Paralichthys olivaceus. For most fish pathogens, including S. parauberis, there are no analytical criteria to distinguish antibioticsusceptible strains from antibiotic-resistant strains. In this study, epidemiological cut-off ($CO_{WT}$) values were generated to classify 75 strains of S. parauberis isolated from 1999 to 2018 as wild type (WT) and non-wild type (NWT) using disc diffusion data and normalized resistance interpretation (NRI) analysis. The susceptibility of the isolates to 16 antibiotics was evaluated using CLSI guideline M42-A. The wild-type cut-off values for amoxicillin, erythromycin, oxytetracycline, and florfenicol for S. parauberis were ${\geq}35$, 31, 28, and 27 mm, respectively. The NWT ratios of S. parauberis strains to treatment with GEN, FFC, ENR, SXT, EFT, VAN, and CHL were 17% or less, indicating that these antibiotics may be used to treat streptococcosis caused by S. parauberis. For recent S. parauberis isolates, the NWT ratios for AMX, ERY, OTC and FFC are much higher than for strains isolated from 1999-2007. The $CO_{WT}$ data from this study will assist aquatic animal disease professionals in prescribing appropriate antibiotics for the treatment of streptococcosis caused by S. parauberis, which will help reduce the misuse and abuse of antibiotics in the aquaculture sector.

• Journal of Life Science
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• v.29 no.1
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• pp.97-104
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• 2019

Streptococcus iniae is a typical fish pathogen causing streptococcosis and it can also cause zoonotic infectious diseases. We studied S. iniae FP5228 isolated from infected olive flounder in Wando, Korea. In a study to find virulence factors in FP5228, we found that the number of live bacteria decreased dramatically in culture medium containing S. iniae FP5228 for more than 24 hr. This phenomenon was hypothesized to be related to Toxin ${\zeta}$ and Antitoxin ${\varepsilon}$ genes, components of the Toxin/ Antitoxin (TA) system on the 14 kb plasmid of FP5228. We used a protein overexpression system to identify it. The pBP1140 vector system was constructed to regulate the expression of Toxin ${\zeta}$ and Antitoxin ${\varepsilon}$ by IPTG and Arabinose. E. coli/pBP1140 strain grew slowly in early growth under toxin expression condition, and it was confirmed by microscopic observation that the strain became longer. S. iniae CK287, lacking a 14 kb plasmid of S. iniae FP5228 strain, was constructed. CK287 bacterial cells did not show rapid killing during culture, and the ability to produce biofilm was also decreased, and toxicity was weakened in cytotoxicity test and fish test. These results suggest that the TA system is involved in physiological regulation and expression of virulence factors in S. iniae FP5228.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.56 no.4
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• pp.380-387
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• 2023

Stocking density is associated with Paralichthys olivaceus growth; thus, fish should be rapidly reared at high densities for commercial reasons. Studies have reported that high stocking density retards growth; however, few have investigated the malformations caused by stocking density stress. This study compared the growth and malformation rates of P. olivaceus at different densities and stress- and immune-related gene expression between malformed and normal fish. Forty days post-hatching, fish (total length, 1.49±0.02 cm) were reared at 800 (low density; LD), 1500 (medium density; MD), and 4000 (high density; HD) fish/m², and the growth rate was measured weekly. On day 30, RNA was extracted from the kidneys, and the expression of stress-, immune-, and malformation-related genes was analyzed using qRT-PCR. The malformation rate in the HD groups was approximately three times higher (62%) than that in the LD and MD groups (approximately 20%), and growth was lower regarding length and weight. The stress-related (HSP70 and GPX) and immune-related (PIR and IgM) genes showed higher mRNA expression in the HD group and malformed fish than in the LD group and normal fish. However, TLR3 showed the opposite results. In summary, high stocking density suppressed growth and increased malformation risk in P. olivaceus.

• Korean Journal of Environmental Biology
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• v.21 no.1
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• pp.26-30
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• 2003

Cytochrome P450 (CYP) induction was determined in microsomes of three aquacultured fish species (Sebastes schlegeli, Paralichthys olivaceus and Pagrus major) and two wild fish species (Mugil cephalus and Stephanolepis cirrhifey) in vitro exposed to $\beta$-naphthoflavone (BNF). The microsomes of five fish were exposed to BNF (5 mM or 10 mM) in dimethylsulfoxide at $30^{\circ}C$ for 9 hr. The CYP contents in most fish increased according to exposure duration for 3 or 5 hour, and then decreased, while steady increase of CYP was observed in P. major for 9 hour. The induction of CYP contents in aquacultured fish species (207~422%) were higher than those in wild fish species (206~207%).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.4
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• pp.339-347
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• 2000

An algicidal bacterium, Micrococcus sp. LG-5 against the harmful dinoflagellate, Cochlodinium polykrikoides was isolated. The optimal conditions for the highest algicidal activity of bacterial culture filtrate showed in the range of $20{\~}30^{\circ}C$, at pH 7.0 and $3.0{\%}$ of NaCl concentration. In addition, $IC_(50)(mean of 50{\%} inhibitory concentration)$ of the culture filtrate against C. polykrikoides after incubation of 5 days was $0.482{\%}$. To investigate heat and pH stability of the culture filtrate of Micrococcus sp. LG-5, the culture filtrate ($pore size, 0.1 {\mu}m$) was heated to $121^{\circ}C for 15 min$ and adjusted pH from 2.0 to 10.0. There were no significant changes in algicidal activity by heat treatment and the pH change between pH from 5.0 to 10.0. The algicidal substances produced from Micrococcus sp. LG-5 were mainly detected in the fraction of $10,000{\~}1,000$ MWCO (molecular weight cut-off). The culture filtrate of Micrococous sp. LG-5 showed antimicrobial activity against Enterococcus faecalis, Escheiichia coli, Uebsiella pneunioniae and Vibrio altinolyticus, but did not show against Pseudomonas aeminosa, P. Buorescens, Salmonella typhi, Staphylococcus aureus, V. cholerae and V parahaemolyicus. The culture filtrate of Micrococcus sp. LG-5 was examined against 16 phytoplankton species and showed the algicidal activity against Ajexandzium tuarense, Eutreptiella Drnnastin, Gymnodinium catenatum, G. mikimotoi, G. sanguineum, eyodinium impuaicum, Heterocapsa triquetra, Heterosipa akashiwo, Prorocentrum micans and Pyraminonas sp.. However no algicidal effects of Micrococcus sp. LG-5 were observed against Chlamydomonas sp., Cylindrotheoa closterium, P. mininum, P. triestimum, Pseudonieschia sp. and Sczipuiella trochoidea. On the other hand, algicidal activity on the tested marinelivefood was not detected except for Isochrysis galbana. In addition, physiological responses of cultured olive flounder (Paralichthys oliraceus) exposed to $1 and 10{\%}$ of the culture filtrate of Micrococcus sp. LG-5 were measured. There were no clear changes in AST, GGT, creatinine, urea, total cholesterol, total protein, albumine, $Mg^(+2), Ca^(+2), Na^+, K^+, and Cl^-$. These results indicate that olive flounders were not affected when they were exposed to the culture filtrate of Micrococcus sp. LG-5.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.5
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• pp.465-472
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• 2001

The effects of water level reduction in rearing tank and fish transference during fish selection process on the stress response (hematological factors, cortisol, glucose, lactic acid and osmolality) of tank-reared olive flounder Paralichthys olivaceus of large (FL), small (FS) and Japanese croaker, Nibea japonica (JC) were examined in running seawater culture system. The water level of rearing unit was lowered from 33 cm to 8 cm in the course of 2 minutes in the water level reduction experiment. The fish were removed from rearing tank (12 ton) to 450 L tank in 30 seconds after capture in the fish transference experiment, In water level reduction, the hematocrit of FL was significantly increased from $14.6\%$ at beginning to $23.5\%$ after 10 hours, However, it decreased to the value of beginning after 46 hours. Plasma cortisol concentration of FL was the highest concentration (13.7 ng/mL) after 22 hours, but it decreased to 4.0 ng/mL at the end of experiment. Cortisol concentration of FS did not show any significant difference during the experiment. The cortisol concentration of JC were significantly higher at 4 hours (282.3 ng/mL) and 22 hours (350.5 ng/mL), Glucose concentration of JC was the highest (138.0 mg/dL) at 22 hours. Lactic acid concentration was not different between experimental groups. In the fish transference experiment, red blood cell of FL was increased from $1.9\times10^6\;cell/{\mu}\;L\;to\;4.2\times10^6\;cell/{\mu}L$ in 24 hours. Blood hemoglobin of JC were significantly elevated in 24 hours. At 1 hour after transference, plasma cortisol concentrations in both fish species were increased to 95.3 ng/mL in FL and 175.5 ng/mL in JC. Glucose concentration of JC was increased to 132.5 mg/dL at 1 hour, 129.5 mg/dL at 3 hours after transference.

• Journal of fish pathology
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• v.19 no.3
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• pp.207-214
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• 2006

Diagnostic monitoring in fish farms with land-based tanks and net cases was conducted in eastern, western, southern and Jeju island of Korea for the summer period from 2000 to 2006. Total 3,518-fish samples of marine and freshwater fishes in 25 fish species were tested for pathogens. Fish species tested were olive flounder (Paralichthys olivaceus), fleshy prawn (Fenneropenaeus chinensis Osbeck), black rockfish (Sebastes schlegeli), rock bream (Oplegnathus fasciatus), red sea bream (Pagrus major), black seabream (Acanthopagrus schlegeli), sea bass (Lateolabrax japinicus), gray mullet (Mugil cephalus), rainbow trout (Onchorhynchus mykiss) and others. The infection rates by bacterial pathogens in the years of 2000, 2001, 2002, 2003, 2004, 2005 and 2006 were 22.4%, 34.5%, 14.1%, 15.3%, 17.7%, 13.5% and 5%, respectively. The infection rates by parasitic pathogens were 20%, 33.8%, 12.4%, 14.1%, 9.2%, 10.5% and 10.7%, respectively. The infection rates by viral pathogens were 22.4%, 13.5%, 10.3%, 5.4%, 9.7%, 10.2% and 15.8%, respectively. The infection rates by mixed pathogens were 10.3%, 0%, 44.9%, 50.9%, 31.9%, 38.4% and 39.6%, respectively. The rates of mixed infections were very low until 2001. The rates were higher than those of singer infections from 2002 to 2006. During the diagnostic monitoring from 2000 to 2006, the main bacterial pathogens were Vibrio (41.2%) and Streptococcus (28.8%). The infection rate by protozoa (85.7%) mainly including Scuticociliates and Trichodina was highest. The infection rate by viral necrosis virus (VNNV, 42.2%) was the highest of the viral pathogens.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.250-256
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• 2017

The prevalence of infections due to pathogenic bacteria such as Edwardsiella tarda, Streptococcus parauberis, and Photobacterium phosphoreum in fish farms in Jeju Island and their management by marine-derived biomaterials was studied. In this study, we isolated eight spices type of marine-derived biomaterials from four sea areas of Jeju Island. An antibiotic disc susceptibility test confirmed that the isolated marine-derived biomaterials showed weak resistance only to oxytetracycline and penicillin and sensitivity to the other antibiotics tested, and antimicrobial activity against fish pathogens with the inhibitory zone of 22 mm, 18 mm, and 19 mm for MD-02, MD-04, and MD-06 against E. tarda strains, respectively, and 19 mm, 22 mm, 30 mm, and 29 mm for MD-01, MD-02, MD-04, and MD-06 against S. parauberis strains, respectively, while all the marine-derived biomaterials showed antibacterial activity against P. phosphoreum. Among the eight biomaterials selected, Bacillus subtilis MD-02 displayed the greatest antibacterial activity against the three tested fish pathogens and also displayed susceptibility to antibiotics. The growth of Bacillus subtilis MD-02 was greatest with the carbon source, dextrine; nitrogen source, peptone; and mineral source, $MgSO_4{\cdot}7H_2O$. Hence, the present study confirmed that the isolate B. subtilis MD-02 from Jeju Island could be a potential antimicrobial agent against fish pathogens and a potential pharmacotherapeutic agent.

• Journal of Korean Society of Environmental Engineers
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• v.34 no.12
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• pp.810-820
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• 2012

Polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) contents in marine sediment and edible fish (4 species) from the estuary near the Yongho wharf in Busan were determined to assess their presence and their potential health risk to the local population through fish consumption was also assessed. Levels of ${\Sigma}PCBs$ and ${\Sigma}DDTs$ in marine sediment were 3.22~197.65 and 1.77~20.27 ng/g dw, respectively. HCHs and endosulfan sulfate concentrations in bottom sediment were 1.42~6.08, 0.56~13.89 ng/g dw, respectively. The bottom sediment in the offshore of the Yongho wharf may be characterized as semi-polluted status with potential adverse marine biological effects in terms of sediment quality guidelines of US NOAA. The mean concentrations of ${\Sigma}PCBs$ in the tissues of olive flounder (Paralichthys olivaceus), Korean rockfish (Sebastes schlegelii), file fish (Stephanolepis cirrhifer) and abbysal searobin (Lepidotrigla abyssalis) were 67.37, 10.20, 48.26, 22.81 ng/g lw, respectively. DDTs and HCHs were also detected in all fish samples. Lifetime cancer risk and target hazard quotient to local residents due to those fish consumption were found to pose negligible cancer and non-cancer risk.

• Journal of the Korean Society for Marine Environment & Energy
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• v.16 no.2
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• pp.88-101
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• 2013

The International Convention for the Control and Management of Ship's Ballast Water and Sediments was adopted at 2004 and then various BWMS (ballast water management system) have been developed. In this study, WET (whole effluent toxicity) test with algae (diatom) Skeletonema costatum as primary producer, invertebrate (rotifera) Brachionus plicatilis as 1st consumer and fish (olive flounder) Paralichthys olivaceus as predator, chemical analysis and ERA (environmental risk assessment) were conducted to assess the unacceptable effect on marine ecosystem by emitting the discharge water treated with AquaStar$^{TM}$ BWMS using electrolysis as main treatment equipment for removing the marine organisms in the ship's ballast water. The most sensitive test organism on discharge water treated with AquaStar$^{TM}$ BWMS was S. costatum that gave the NOEC value of 25.00%, LOEC value of 50.00% and 72hr-$EC_{50}$ value of 69.97% from WET test result for 20 psu salinity treated discharge water. NOEC and LOEC value of B. plicatilis and P. olivaceus exposed at 20 psu salinity treated discharge water were 50.00% and 100.00%, respectively. In the chemical analysis results, total number of substances produced by AquaStar$^{TM}$ BWMS was 18 which were bromate, 7 volatile halogenated organic compounds, 7 halogenated acetic acids, 3 halogenated acetonitriles and chloropicrin. Eighteen substances did not consider as persistence and bioaccumulative chemicals. Uncertainty of toxic property of 18 substances was high. PECs of 18 substances calculated by MAMPEC model were ranged from $4.58{\times}10^{-4}$ to $4.87{\mu}g\;L^{-1}$, PNECs of them were ranged from $1.6{\times}10^{-2}$ to $3.2{\times}10^2{\mu}g\;L^{-1}$. And, the PEC/PNEC ratio of 18 substances did not exceed 1. Therefore, ERA for produced substances indicate that the discharge water treated with AquaStar$^{TM}$ BWMS does not pose unacceptable effect on marine life. And $EC_{50}$ value of S. costatum on discharge water treated by BWMS using the electrolysis had positive correlation with initial TRO concentration, concentration and kind & level of HAAs.