• Asian-Australasian Journal of Animal Sciences
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• v.23 no.7
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• pp.916-923
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• 2010

We report nutritional physiology and non-specific immune responses of vitamin E in parrot fish for the first time. This study aimed to investigate the essentiality and requirements in diets based on growth performances, non-specific immune responses and a challenge test against Vibrio angullarum. Six casein-gelatin based semi-purified diets were formulated to contain six graded levels of DL-${\alpha}$-tocopheryl acetate (${\alpha}$-TA) at 0, 25, 50, 75, 100 and 500 mg/kg diet (designated as E0, E25, E50, E75, E100 and E500, respectively) and fed to triplicate groups of juvenile parrot fish for 12 weeks. The analyzed dietary concentrations of vitamin E were 0, 38, 53, 87, 119 and 538 mg/kg diet for E0, E25, E50, E75, E100 and E500, respectively. At the end of the feeding trial, growth performance and feed utilization of fish fed the E25 were significantly higher compared to that of fish fed the other diets. Liver ${\alpha}$-tocopherol concentration was significantly increased with an increase in dietary ${\alpha}$-TA in a dose dependent manner. No apparent clinical signs of vitamin E deficiency and mortality were observed in fish fed the basal diet for 12 weeks. Among the immune responses assayed, phagocytic (NBT assay) and myeloperoxidase activities were significantly increased with increment of dietary ${\alpha}$-TA levels. During the challenge test with V. anguillarum, E75, E100, and E500 diets resulted in higher survivals than E0, E25 and E50 diets. The findings of this study suggest that parrot fish require exogenous vitamin E and the optimum dietary level could be approximately 38 mg ${\alpha}$-TA/kg diet for normal growth and physiology. Dietary ${\alpha}$-TA concentration over 500 mg/kg could be required to enhance the nonspecific immune responses and improve the resistance of juvenile parrot fish against V. anguillarum.

• Journal of Fisheries and Marine Sciences Education
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• v.25 no.6
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• pp.1285-1293
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• 2013

This study was conducted to investigate the effects of dietary inclusion of various additives Solid sulfur, fucoidan, and glucan on growth performance and immune responses in Juvenile olive flounder paralichthys olivaceus. Six experimental diets supplemented with : no additives (CON); 0.5 and 1.0% solid sulfur (S); 0.2 and 0.4% fucoidan (F); 0.1% glucan (G) of these additives diet on a dry-matter basis. Fish averaging $8.2{\pm}0.17g$ were fed one of six experimental diets in triplicate groups for 6 weeks. By the end of the feeding trial, weight gain, feed efficiency, specific growth rate, protein efficiency ratio, hepatosomatic index and condition factor of fish fed diet CON were significantly lower then those of fish fed the all additives diets (P < 0.05). In challenge test, fish were infected by intraperitonel injection of 0.1 ml bacterial suspension with Edwardsiella tarda per fish after the feeding trial. As a result fish fed All additive diet showed a lower cumulative mortality then did fish fed CON diet throughout the challenge test. In conclusion, these results indicated that solid sulfur, fucoidan and glucan enhanced the growth, feed efficiency and non-specific immune activity of juvenile flounder and protect the fish against microbial infections.

• Journal of fish pathology
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• v.15 no.2
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• pp.65-75
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• 2002

This study was performed to know the effects of stress induced by the daily fluctuation of water temperture from 18$^{\circ}C$ to 25$^{\circ}C$ up and down for 30 days on the defence mechanism of olive flounder. Puralichthys olivaceus. To make clear the temperature stress on the defense mechanism of the tested fish. several factors of immune response such as counting of leucocyte appearance in peripheral blood, phagocytic activity in whole blood cells, nitroblue tetrazolium(NBT) reduction, chemiluminescence(CL) response, and lysozyme activity were investigated at 28 days after giving the change of water temperature. The fish was controlled under the none feeding condition during experimental period. Mortality of the tested fish was rapidly increased up to 22% within the first one week of the experimental period without any additional stress factors. The number of neutrophil of peripheral blood in the tested group was significantly higher than the control group at the 2nd week. but the number of lymphocyte was significantly lower than the control group at the 1st and 3rd day of the experimental period. respectively. In the NBT reduction test, the activity of macrophage in the control group fish was the highest on the 7th day while that in the tested group was on the 3rd day. Also. the phagocytosis of tested group against formalin killed cells was retarded compared with the control. CL response of the tested group was significantly lower from 2nd to 5lh day of the experimental period than the control. 'The lysozyme activity of tested group was remained higher during the experimental period than the control. Even though the tested fish showed different results in some non-specific factors of immune respceses between tested and control group fish, olive flounder seems highly adaptable in repealed water temperature change in condition after one week under the given temperature fluctuation range.

• Journal of fish pathology
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• v.22 no.1
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• pp.45-52
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• 2009

Zebrafish was firstly applied to an experimental model for scuticociliatosis caused by Philasterides dicentrarchi, a facultative parasitic ciliate in cultured marine fish. The susceptibility of zebrafish to infection of P. dicentrarchi was assessed by intraperitoneal injection of the ciliates, which produced typical symptoms of scuticociliatosis and significant mortality. The potential use of zebrafish as a model to evaluate the vaccine efficacy against scuticociliatosis was analyzed by immunization of zebrafish with the ciliates lysate. Furthermore, the effect of different adjuvants, such as Quillaja saponin (QS), Montanide, and Freund’s incomplete adjuvant (FIA) on the protective efficacy of the vaccine was investigated. Groups of zebrafish injected with QS or Montanide alone showed higher survival of fish against challenge test compared to control fish. The results suggest that adjuvant-mediated enhancement of innate immune responses play important roles in protection of fish against scuticociliatosis. The considerably high survival in the fish immunized with the antigen alone indicates that the ciliate lysate itself is highly immunogenic to zebrafish, which can elicit protective immune responses. The protective potential of the antigen, ciliate lysate, was enforced through combined administration with adjuvants including QS, Montinide and FIA. No or low mortalities in the groups of fish immunized with the antigen plus adjuvants suggests that the adaptive immune responses of zebrafish might be accelerated by the adjuvants or the protective potential of the antigen and adjuvants might synergistically interact. In spite of several shortcomings such as difficulties in sampling of serum and leucocytes enough to routine immunological analyses, zebrafsih might be the most convenient experimental animal for scuticociliatosis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.55 no.5
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• pp.688-696
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• 2022

Obesity could cause immune-physiological disorders in fish. Yet, little is known about the impact of obesity on stress and histological responses. This study aimed to determine histological and physiological changes in and vulnerability of overfeeding-induced obese rainbow trout Oncorhynchus mykiss exposed to stress condition. Control, intermediate and overfed groups were fed at 1.5, 2.5 and 3.09% of their body weight per day, respectively, for eight weeks. Weight gain, body mass index, hepatosomatic index and serological parameters, and histology of liver were measured in five fish from each group at week 0, 2, 4, and 8. At week 8, 20 fish from each group were exposed to heat stress by increasing water temperature at a rate of 3℃ per day from 15 to 25℃ and maintaining the final temperature for 10 days. Overall, overfed fish showed significantly higher weight gain, body mass index, and serological parameters than those of fish in the other groups. Fish in the overfed and intermediate groups displayed multifocal infiltration of inflammatory cells in hepatic parenchyma. Mortality rate and serological parameters of fish in the overfed group exposed to heat stress were significantly higher than those of fish in the other groups, indicating increased vulnerability to environmental stress.

• Journal of fish pathology
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• v.28 no.1
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• pp.1-7
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• 2015

In 2009 outbreaks of an emerging disease in shrimp farms were first reported in China. The disease was known as early mortality syndrome (EMS), or acute hepatopancreatic necrosis disease (AHPND). Since 2009, the disease has been spread to Vietnam, Malaysia, Thailand and Mexico. The causative agent of the disease was identified as Vibrio parahaemolyticus. It is a common seawater inhabitant bacterium, and the pathogen can sometimes contaminate seafood. The disease has caused mass mortality of cultivated shrimp, and huge economic losses in the countries named above. In order to prevent the introduction and establishment of AHPND, emergency measures, such as strengthening of import conditions and even import bans, were put in place by many other countries. In Korea, on the other hand, there are large quantities of shrimp imports from countries, such as Thailand and Vietnam. Transportation of live and fresh dead shrimp is highly likely, and could be a transmission pathway if the shrimp are sourced from populations in AHPND endemic areas. It is important to recognize that importing countries may provisionally adopt sanitary or phytosanitary measures on the basis of available pertinent information, including that from the relevant international organizations, as well as from sanitary or phytosanitary measures applied by other countries based on "Agreement on Application of Sanitary and Phytosanitary Measures". It is pertinent that Korea also takes proper emergent measures to keep out diseases and provide safe seafood.

• Journal of fish pathology
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• v.23 no.3
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• pp.313-322
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• 2010

Edible tunicate Halocynthia roretzi, one of the most commercially important aquatic organisms in Korea, has been killed by tunic softness syndrome since last decade. The intracellular protistan parasite observed by the transmission electron microscope in hemocytes of the tunicate was considered to be the causative agent of the mass mortality. The goal of the present work is to examine the characteristic features of the parasite by identifying the 18S rDNA sequences of the parasite. The experiments conducted include amplification of presumptive 18S rDNA from diseased tunicate tissues with UNonMet-PCR and sequencing the product. A preliminary phylogenetic analysis was performed on the presumptive parasite rDNA. A digoxigenin labeled DNA probe was designed on the basis of the sequences of rDNA. Dig-ISH assay was conducted to diagnose the protistan parasite. A PCR using UNonMet-PCR primer generated 595 bp SSU rDNA fragment. Subsequently, PCRs with primer pair expended this sequence to 1542 bp. This is the first partial sequences of SSU rDNA gene to be published on the protistan parasite that has presumed causing the mass mortality of tunicate. Since the Dig-ISH technique demonstrated the presence of infection in hemocytes on the all host tissues, the fragment was confirmed to be the intracellular protistan parasite SSU rDNA. A phylogenetic analysis suggested that the protistan parasite may be a unique eukaryote that is closely related to Apicomplexa.

• Journal of fish pathology
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• v.35 no.2
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• pp.141-155
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• 2022

Rock bream iridovirus (RBIV) causes high mortality and economic losses in the rock bream (Oplegnathus fasciatus) aquaculture industry in Korea. Viral open reading frames (ORFs) expression profiling at different RBIV infection stages was investigated using microarray approaches. Rock bream were exposed to the virus and held for 7 days at 23 ℃ before the water temperature was reduced to 17 ℃. Herein, 28% mortality was observed from 24 to 35 days post infection (dpi), after which no mortality was observed until 70 dpi (end of the experiment). A total of 27 ORFs were significantly up- or down-regulated after RBIV infection. In RBIV-infected rock bream, four viral genes were expressed after 2 dpi. Most RBIV ORFs (26 genes, 96.2%) were significantly elevated between 7 and 20 dpi. Among them, 12 ORF (44.4%) transcripts reached their peak expression intensity at 15 dpi, and 14 ORFs (51.8%) were at peak expression intensity at 20 dpi. Expression levels began to decrease after 25 dpi, and 92.6% of ORFs (25 genes) were expressed below 1-fold at 70 dpi. From the microarray data, in addition to the viral infection, viral gene expression profiles were categorized into three infection stages, namely, early (2 dpi), middle (7 to 20 dpi), and recovery (25 and 70 dpi). RBIV ORFs 009R, 023R, 032L, 049L, and 056L were remarkably expressed during RBIV infection. Furthermore, six ORFs (001L, 013R, 052L, 053L, 058L, and 061L) were significantly expressed only at 20 dpi. To verify the cDNA microarray data, we performed quantitative real-time PCR, and the results were similar to that of the microarray. Our results provide novel observations on broader RBIV gene expression at different stages of infection and the development of control strategies against RBIV infection.

• 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.

• Fisheries and Aquatic Sciences
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• v.12 no.4
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• pp.350-353
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• 2009

As a result of human industrial development, carbon dioxide ($CO_2$) is currently accumulating in the atmosphere and dissolving into the oceans. Sequestration into the deep sea has been proposed as a possible solution to this increasing atmospheric $CO_2$, although the impact of such a program on marine ecosystems is unknown. We examined the effects of increased $CO_2$ levels on the growth of the olive flounder, Paralichthys olivaceus. Juvenile olive flounder 40 days post hatching were exposed to two levels of $CO_2$ (3.60-7.55 and 4.05-11.46 kPa) in running seawater for 26 days. During the exposure period, the pH and $CO_2$ levels of the water were measured, and the numbers of dead individuals were counted in each aquarium. Following the exposure period, the total lengths (mm) and body weights (mg) of the juvenile fish were measured. Both $CO_2$ treatments significantly increased fish mortality compared to controls ($19.87\pm4.53%$ vs. 7.14% and $75.96\pm1.36%$ vs. 7.14% for high and low doses, respectively). After the high $CO_2$ treatment, total length ($14.98\pm6.58$ mm vs. $19.52\pm1.83$ mm) and body weight ($28.92\pm13.85$ mg vs. $67.35\pm18.32$ mg) of the exposed flounder were reduced compared to the control fish; however, no significant differences in these values were observed after the low $CO_2$ dose. These results suggested that $CO_2$ exposure inhibits growth in the juvenile stage and that $CO_2$-enriched seawater is toxic in the early life stages of olive flounder.