• Journal of fish pathology
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• v.13 no.2
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• pp.121-127
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• 2000

From August to October 1998, over 60% mortality of cultured striped beakperch Oplegnathus fasciatus was occurred in net cages along the southern coast of Korea. Moribund fish showed some clinical signs of lethargic behavior, dark coloration or decoloration, severe gill anemia and enlargement of spleen. Also enlarged basophilic cells showing Feulgen -positive reaction were observed in the tissue section of spleen, kidney, liver and heart of the diseased fish. GF cells inoculated with spleen homogenate of diseased fish produced cytopathic effect of enlarged and rounded cells, therefore the causative virus was isolated from diseased fish. Striped beakperch fingerlings intraperitoneally inoculated with the causative virus ($10^4TCID_{50}$/0.1 ml) revealed symptoms similar to those of naturally infected fish and died from 7 to 14 days post injection. Transmission electron microscopy revealed that the causative virus was enveloped icosahedral particle with 120~130 nm in diameter. PCR products of the expected size (500 bp) were amplified with a primer set based on the ATPase gene of RSIV(red sea bream iridovirus) using template DNAs which were extracted from the spleen of diseased fish and GF cells inoculated with the causative virus. According to the analysis of nucleotide sequence of these PCR products, the sequence from ATPase cDNA gene of the causative virus showed 95% homology with that of RSIV. These results indicate that the mass mortality in the cultured striped beakperch was caused by the infection of iridovirus similar to RSIV.

• Journal of fish pathology
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• v.9 no.1
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• pp.1-9
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• 1996

Many encysted metacercariae of Clinostomum complanatum(Rudolphi, 1814) were found in the flesh near the skin of goldfish(Carassius auratus) sultured at a fish farm in Chonbuk Province, Korea, in the summer of 1995. The mortality reached about 7% from July 10 to August 20. Encysted and excysted metacercariae were$700\sim877{\times}700\sim775{\mu}m$ and $2.4\sim5.5{\times}0.7\sim1.37{\mu}m$ in size, respectively. The parasites looked yellowish due to the yellow particulate matter in the intestine. Of 276 fish examined, 234(84.8%) were infected with a mean intensity of 216.6(range 1~1.214) parasites. Fish of 2.0~2.5 cm in body length were most frequently(94.3%) infected but those of 3.1~3.5 cm were the least infected(75.27%). The isthmus was most heavily infected. There was no difference in prevalence between two forms of goldfish, Gyariko an Ryugum. The sites of infection were swollen and recognized as yellow spots of 0.4~0.8 mm in diameter. Fish infected with more than 200 metacercariae became moribund with listless swimming and finally died.

• Journal of fish pathology
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• v.4 no.2
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• pp.79-85
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• 1991

In Feb., 1990 an epizootic disease to masu salmon fry, Oncorhynchus masou cultured at the hatchery of rainbow trout in Samchok, Kwangwon-do have broken out and induced high mortality over 70%. Externally, the diseased fish showed dark discoloration, abdominal distension with ascites, slight exophthalmus and fecal casts. Internally, the gill and the liver of diseased fish were edematous and pale, and the stomach of moribund fish contained the milkish fluid. Microscopically there was extensive necrosis of the hematopoietic tissue in kidney and spleen, and scattered necrosis of pancreas, liver and lateral muscle. Especially, the necrosis of lamina propria, muscle layer and tela submucosa in the digestive tract known as the typical signs associated with infectious hematopoietic necrosis virus was seen clearly.

• Journal of Marine Bioscience and Biotechnology
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• v.1 no.3
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• pp.163-169
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• 2006

In the survey of the samples infected by iridovirus, we found the natural outbreak of iridovirus disease in the ornamental fish, pearl gourami (Trichogaster leeri) in Korea. It was characterized by the appearance of enlarged cells and necrosis in the observation of the imprinted spleen cells under iridovirus. To determine the infection of iridovirus more accurately, PCR using 2 different primer sets for MCP and ATPase gene used for the diagnosis of iridovirus infection was done and found the produced fragments matched with the size of expectation. Partially determined nucleotide sequences of the MCP gene of the iridovirus isolated in this study showed very high homology (99.6%) with that of ISKNV as a reference strain. In challenge experiment with the iridovirus isolated from the moribund of pearl gourami, the pathogenicity of the isolated iridovirus was confirmed and suggested the potential of the risk associated the transfer of iridovirus from the ornamental fishes to the marine culturing fishes.

• 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 fish pathology
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• v.11 no.2
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• pp.129-136
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• 1998

In March 1997, a new rhabdovirus was isolated from moribund cultured Japanese flounder Paralichthys olivaceus in sea water tank and cage culture systems in Kyung-Nam and Chun-Nam province, Korea. At temperature $15^{\circ}C$ the virus replicated and induced cytopathic effects (CPE), which progressed to eventual cytolysis, in susceptible cell lines, including RTG-2 and EPC. The CHES-214 cell line was refractory. Virus particles were bullet-shaped and measured $70nm{\times}100$ to 150 nm in size. The isolate was sensitive to pH 3, to diethyl ether, and to heat ($50^{\circ}C$ 5 min, $60^{\circ}C$ 1 min). Viral replication was not inhibited by $10^{-4}$ M 5-iododeoxyuridine. Virus infectivity was reduced by anti-HRV (8401-H) rabbit serum, but can not reduced by antisera against infectious hematopoietic necrosis virus (IHNV), chum salmon reovirus (CSV), retrovirus of salmonid (RVS) and infectious pancreatic necrosis virus (IPNV). HRV virus antigen was detected by fluorescent antibody test (FAT) in the cytoplasm of infected EPC cell. Purified isolates virions were composed of: polymerase (L), glycoprotein (G), nucleoprotein (N) and 2 matrix proteins (M1 and M2). Based upon their relative mobilities, the estimated molecular weights of the proteins were: L, 160 kDa; G, 55 kDa; N, 45 kDa; M1, 26 kDa; and M2, 22 kDa.

• 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 Ichthyology
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• v.29 no.3
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• pp.157-164
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• 2017

In 2015, a nervous necrosis virus (NNV) was isolated from sevenband grouper, Epinephelus septemfasciatus, maintained in land-based aquaculture system at below $12^{\circ}C$ in winter. Mortality was up to 30% in brood fish, over 4 kg of body weight. Moribund fish showed clinical sings typical of viral encephalopathy and retinopathy (VER), also called viral nervous necrosis (VNN), such as uncoordinated, corkscrew-like swimming behavior, belly-up at rest, darkening of body, cloudy eyeball and hyperinflation of the swim bladder. Aetiology of the disease was confirmed by gross observation of clinical signs, histopathology and molecular diagnosis. Histological studies revealed severe vacuolation and necrosis in the brain. Molecular diagnosis by revere transcription-polymerase chain reaction (RT-PCR) specific to batanodavirus yielded a positive result. The nucleotide sequences of the PCR-amplified fragment were 99.48~100% similar to barfin flounder nervous necrosis virus (BFNNV) genotype and most closely aligned with Pacific cod betanodavirus (PCNNV). This is the first report of natural batanodavirus, NNV infection in sevenband grouper reared in low water temperature during winter (below $12^{\circ}C$) in Korea.

• Journal of fish pathology
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• v.10 no.2
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• pp.97-111
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• 1997

In the winter of 1995, mass mortality occurred in cultured flounder, Paralichthys olivaceus in Gurongpo, Kyoungbuk, Korea. From the observations of moribund and dead fish, parasitic ciliates, which were shown as white spots to the naked eye, were considered to be involved in the mass mortality. From heavily infected flounders, histopathological, morphological and biological characterization of these ciliates were carried out. In the histological observation, many ciliates were found under the epithelia of gill filaments and skin, and caused hyperplasia of epithelial and mucus cells at the infected areas. The ciliates found on the body surface, fins and gills were very similar to Cryptocaryon irritans. However the ciliates showed two different patterns of reproductian, i.e., typical form(palintomy)and atypical form(budding plus multiple fission) at $16^{\circ}C$ of water temperature. The occurrence ratio between typical and atypical form was about 3:2. Tomitogenesis takes 8-14 days in the typical and 13-15 days in the atypical form. In the viability test at different temperatures and salinities, the typical form died below 30‰ at $12^{\circ}C$, below 20‰ at $16^{\circ}C$, below 15‰ at $20^{\circ}C$, and below 25‰ at $24^{\circ}C$, respectively. On the other hand, the atypical form died below 20‰ at $12^{\circ}C$, below 15‰ at 16-$20^{\circ}C$, and below 25‰ at $24^{\circ}C$, respectively. The results suggested that the atypical has better viability at low salinity than that of the typical at low temperatures. In the excystment time and success rates of excystment according to temperatures, the typical form showed 8 days, 30% at $12^{\circ}C$ : 6.5 days, 50%, at $16^{\circ}C$ : 5.5 days, 75% at $20^{\circ}C$ : and 7 days, 10% at $24^{\circ}C$, respectively. On the other hand, the atypical form showed 15.5 days at $12^{\circ}C$ : 14 days, 76.6% at $16^{\circ}C$ : 12 days, 72.2% at $20^{\circ}C$ : 10 days 31.6% at $24^{\circ}C$, respectively. The results suggested that the atypical form had longer excystment time than that of the typical form at any temperature and showed better stability at low temperatures.

• Journal of Aquaculture
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• v.20 no.4
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• pp.278-288
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• 2007

Shrimp culture in Korea had been rapidly developed during 1990's and the production of farmed shrimp reached 3,268 mt from 2,605 ha in 2001. However the shrimp production decreased to 2,368 mt in 2004 because of the mass mortality due to outbreak of white spot syndrome virus (WSSV). WSSV which is one of the most serious threats associated with cultured shrimp around the world has given the 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. Various polyculture technologies of shrimp with shellfish, finfish or seaweeds have been implemented to reduce economic damages by mass mortalities of shrimp. Among them, the polyculture of shrimp with carnivorous fish can suppress or delay the viral outbreak of shrimp ponds because the fish may selectively eat the moribund shrimps infected by virus. To determine the selective predatory effect of river puffer Takifugu obscures on WSSV infected shrimp, postlarvae of Litopenaeus vannamei and Fenneropenaeus chinensis. One-year old river puffers were stocked to four earthen ponds of $1,616-1,848\;m^2$ in surface area as followings: polyculture LvP, L. vannamei ($43.4/m^2$)+puffer ($0.22/m^2$); control Lv, L. vannamei ($46.9/m^2$); polyculture FcP, F. chinensis ($30.3/m^2$)+puffer ($0.25/m^2$); control Fc, F. chinensis ($24.6/m^2$). Ponds of control Fc and polyculture FcP had mass mortalities by WSSV outbreak on the $51^{st}$ and $57^{th}$ days of culture respectively. The shrimps of polyculture LvP and control Lv were harvested on the $95^{th}\;day$. Shrimp survival rates of polyculture LvP and control Lv were 32.4% and 18.2% respectively and shrimp productivity of polyculture LvP was 69.2% higher than that of control Lv. Concentration of nutrients (TAN, $NO_2-N$, $NO_3-N$) was maintained within optimal ranges for shrimp growth although that of polyculture ponds showed at least two times higher than that of control ponds. The results suggest that polyculture of L. vannamei with river puffer is higher than monoculture in survival rate and productivity. In addition, F. chinensis should be carefully cultured because this species shows much higher susceptibility to WSSV than L. vannamei.