• Asian-Australasian Journal of Animal Sciences
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• v.25 no.8
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• pp.1184-1189
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• 2012

White spot syndrome virus (WSSV) is one of the major viral pathogens affecting shrimp aquaculture. Four proteins, WSSV199, WSSV 222, WSSV 249 and WSSV 403, from WSSV are predicted to encode a RING-H2 domain, which in presence of ubiquitin conjugating enzyme (E2) in shrimp can function as viral E3 ligase and modulate the host ubiquitin proteasome pathway. Modulation of host ubiquitin proteasome pathway by viral proteins is implicated in viral pathogenesis. In the present study, a time course expression profile analysis of WSSV Open Reading Frame (ORF) 199 and Penaeus monodon ubiquitin conjugating enzyme (PmUbc) was carried out at 0, 3, 6, 12, 24, 48 and 72 h post WSSV challenge by semi-quantitative RT-PCR as well as Real Time PCR. EF1${\alpha}$ was used as reference control to normalize the expression levels. A significant increase in PmUbc expression at 24 h post infection (h.p.i) was observed followed by a decline till 72 h.p.i. Expression of WSSV199 was observed at 24 h.p.i in WSSV infected P. monodon. Since the up-regulation of PmUbc was observed at 24 h.p.i where WSSV199 expression was detected, it can be speculated that these proteins might interact with host ubiquitination pathway for viral pathogenesis. However, further studies need to be carried out to unfold the molecular mechanism of interaction between host and virus to devise efficient control strategies for this chaos in the shrimp culture industry.

• Journal of fish pathology
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• v.22 no.1
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• pp.15-21
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• 2009

White spot syndrome virus (WSSV) is a strong causative agent for high mortality in cultured and wild shrimps. From this study, the WSSV prevalence in marine organisms around shrimp farm as well as live feed-mediated transmission of WSSV to farmed shrimps were investigated. Based on nested-PCR method, WSSV was detected in wide array of marine organisms including Perinereis aibuhitensis (81.3% of prevalence rate, 13/16), Enedrias fangi (100%, 16/16), Ruditapes philippinarum (20%, 2/10), crab larvae (100%, 10/10), copepoda (30%, 3/10), Periophthalmus modestus (50%, 5/10), Pachygrapsus crassipes (10%, 1/10), Helice tridens (20%, 2/10) and Neomysis sp. (70%, 7/10). On the other hand, WSSV was not detected in Bullacta exarata, Uca arcuata, and Reishia clavigera. The percent prevalence of WSSV in wild shrimps, Fenneropenaeus chinensis was only 6%, but markedly increased up to 56% after a feeding trial using polychaete worms for one month, indicating that the live feed is one of significant carriers of WSSV to shrimps under practical farming conditions.

• 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 Aquaculture
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• v.20 no.4
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• pp.270-277
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• 2007

White spot syndrome virus (WSSV) which is the most serious threat to cultured shrimp around the world has given enormous economic damages to shrimp culture industry every year since it was found from the shrimp ponds in the west coast of the South Korea in 1993. WSSV has strong infectivity as well as virulence and it can be rapidly transmitted among shrimps in ponds by cannibalism of infected ones. Polyculture of shrimps with carnivorous fish has been applied in commercial shrimp farms to suppress or delay the viral outbreak because the fish may selectively eat the moribund shrimps infected by virus. To determine the selective predatory effect of a carnivorous fish, river puffer Takifugu obscurus on white shrimp Litopenaeus vannamei, polyculture trials in laboratory scale of WSSV-infected and non-infected shrimps with river puffer were conducted in concrete round tanks of $28.26\;m^2$ in surface area as followings: 1) juvenile shrimps (B. W. 0.62 g) with 5 months old puffer (B. W. 11.60 g) cultured for 8 days, and 2) sub-adult shrimps (B. W. 6.84 g) with 16 months old puffer (B. W. 85.82 g) cultured for 5 days in order to know the effects according to size difference of cultured animals. In polyculture of juvenile shrimp with 5 months old puffer, survival rates of infected and non-infected shrimps were 46.0% and 89.1% respectively and in that of sub-adult shrimp with 16 months old puffer those were4% and 48% respectively. The results showed that puffer tends to selectively prey on virus infected shrimps among infected and non-infected ones in a limited space with although there is difference in predatory rate with age and density of animals. Regardless of different densities and ages of animals as well as health condition of shrimps, however, there were low differences in daily biomass of shrimp consumed per kg body weight of puffer. This finding suggests that puffer preys on healthy shrimps when moribund shrimps were not sufficient. Therefore, farmers should consider the total biomass of puffer as well as density and stocking time when they stock puffer into shrimp ponds for polyculture.

• Journal of Microbiology
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• v.40 no.4
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• pp.295-300
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• 2002

Previously, we reported induced expression of developmentally regulated CTP-binding protein 2 (DRG2) in fish cells at the late stage of rhabdovirus infection. To investigate the biological role of fish DRG2 (fDRG2), we transfected CHSE-214 cells with an expression vector containing complete fDRG2 fused to the N-terminal end of an enhanced green fluorescent protein (EGFP). Low level expression of fDRG2-EGFP did not induce morphological change or cell death. However, a high level expression of fDRG2-EGFP induced cell rounding and caused depletion of the cell population in FACS analysis. Several truncated fragments were fused to EGFP. FACS analysis was conducted to determine the presence of cells expressing high levels of the resulting chimera. While cells expressing a high level of N-terminus were detected, those expressing high levels of the C-terminal fragment 243-290 containing the G4 motif were absent in FACS analysis. Based on these observations, we propose that overexpression of fDRG2 may induce cell rounding, a representative cytopathic effect of virus-infected cells in the late stage of infection and the C-terminus of the fDRG2 is essential for this function.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.4
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• pp.352-358
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• 2011

The presence of ISKNV-like viruses in various freshwater ornamental fish species imported from Asia was confirmed by polymerase chain reaction(PCR) amplification of the ATPase(adenosine triphosphatase) gene. Interestingly, molecular analyses of the Open Reading Frame 25(ORF25) region of these isolates based on the ISKNV(Infectious spleen and kidney necrosis virus) genome revealed the presence of various repetitive sequences. ORF25 repeat sequence length had no effect on cumulative mortality of rock bream Oplegnathus fasciatus challenged with tissue homogenates of infected pearl gourami, Trichogaster leeri; silver gourami, Trichogaster microlepis; blue gourami, or Trichogaster trichopterus. All isolates induce cumulative mortalities after 12 days of infection, confirming that ORF25 polymorphism did not affect the pathogenicity of ornamental fish megalocytiviruses that cross infect rock bream, a seawater fish. Also, no statistically significant differences in spleen index or viral copy number in infected tissues was detected between isolates with varying ORF25 repeat sequence lengths. However, further studies are necessary to fully characterize the functional characteristics of these polymorphisms in megalocytivirus disease in ornamental fishes.

• Journal of fish pathology
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• v.17 no.2
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• pp.75-82
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• 2004

Interferons are kinds of cytokines that play an important role in antiviral defense mechanisms during viral infection by converting cells to synthesize proteins that inhibit viral replication. In the present study, an antiviral response against Spring viremia of carp virus (SVCV) was developed in common carp, Cyprinus carpio following stimulation with the potent interferon inducer, poly inosinic : cytidylic acid (poly I:C). Poly I:C injected fish showed lower cumulative mortalities against SVCV than untreated fish did. Moreover, in vitro study showed that head kidney leucocytes (HKLs) produced an interferon-like cytokine (ILC) after stimulation with poly I:C. According cytopathic effect reduction (CPER) assay to examine antiviral activity of crude ILC, the capacity of HKLs for ILC production was highest at 1×$10^6$cells/ml and significantly enhanced when treated with 20~50$\mu{g}$/ml of poly I:C. The optimal temperature and concentration of FBS to induce the highest ILC production were $20^\circ{C}$ and 5%, respectively.

• Journal of fish pathology
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• v.21 no.1
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• pp.35-43
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• 2008

Pathological symptoms and hematological parameters of red sea bream, Pagrus major and water temperature in the culture ground was investigated to clarlify the cause of winter moratility. Dead fish showed green liver and accumulation of ascites in the cavity. A few Bivagina tai were also found on the gill but either bacteria or virus were not. When hematological parameters from fish taken before/after the winter mortality were compared, blood glucose, serum AST and ALT, and total cholesterol, trigyceride and total protein were significantly decreased in the fish after the winter mortality. These results may explain that the nutritional level of fish was decreased because fish could not fed during winter season. According to the CORI monitoring system operating by KODC, NFRDI a long term water temperature from Dec. 25, 2005 to Feb. 24, 2006 (60 days) were exposed the low water temperature environments to the red sea bream. First of all, mass mortality began at Sarayngdo where low temperature below 8℃ continued for 42 days. The winter mortality did not occured in the depths of 9 m to 19.2 m where difference of water temperature in the surface and bottom was only within 0.1℃. But in the depth of 7.5 m to 11 m winter mortality occured where water temperature in the surface and bottom showed much variation ranged from 0.2℃ to 1.4℃. From these results, great difference of water temperature in the surface and bottom of the culturing area might results in winter mortality of red sea bream.

• Journal of fish pathology
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• v.28 no.1
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• pp.27-35
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• 2015

VHSV is a major viral agent that affects freshwater and marine fish, causing serious economic losses in aquaculture in the world. Due to their filter-feeding activity, bivalve mollusks may act as viral transmitters after accumulation of the fish viruses released into seawater from infected fish. Amplification by RT-PCR was carried out to investigate the presence of VHSV in pacific oysters (Crassotrea gigas) and blue mussels (Mytilus edulis), inhabiting regions around aquatic farms in Korea. Primers designed from conserved regions of VHSVs allowed us to detect four different types of VHSV in a single PCR. Twenty two of the eighty four samples showed positive results of VHSV in a 2-step RT-PCR. Using six positive samples from three different regions in Korea, we cloned and sequenced the glycoprotein (G) gene (467-bp long) of VHSVs. Genetic analysis of the VHSVs detected in shellfish in various geographical areas of Korea showed highly restricted results to VHSV type Iva. This was in agreement with the reports showing only a single genotype of VHSV (Iva genotype) in outbreaks in cultured or wild fish in Korea. Consequently, we investigated VHSVs carried by bivalve mollusks inhabiting the vicinity of aquatic farms, and revealed correlationship between the type of viral accumulated in shellfish by filter-feeding, and those detected in disease outbreaks in fish.

• Journal of Fisheries and Marine Sciences Education
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• v.25 no.6
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• pp.1315-1323
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• 2013

The purpose of this research is to evaluate the effects of a commercial mixed extruded pellet(EP) and raw fish moist pellet(MP) diet on diseases prevalence and serological constituents and HSI of korean rockfish, Sebastes schlegeli grown in two commercial-scale aquaculture farms from June to December in 2009. The growth of MP-fed rockfish was slightly faster than EP-fed rockfish during experimental period, but did not show significant differences. The levels of TCHO, TG, Glucose and TP in the MP-fed rockfish group were higher than those of EP-fed group, whereas there was no significant difference in other biochemical factors of blood between two groups. In addition, bacteria and virus were not detected in both MP- and EP-fed rockfish groups, and the infection rates of gill fluke between two groups did not show clear difference. In conclusion, there was no comparable difference in growth and the health extents of one year rockfish by administration of the commercial mixed extruded pellet(EP) and raw fish moist pellet(MP) diets.