• 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.

• Fisheries and Aquatic Sciences
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• v.11 no.4
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• pp.195-204
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• 2008

Viral diseases are major emerging problems of shrimp that have affected the production, and even complete losses for shrimp farms. In this study, we developed a sensitive TaqMan real-time PCR method to quantify white spot syndrome virus (WSSV) and hepatopancreatic parvovirus (HPV) in the shrimp and pond water in which fleshy shrimp, Fenneropenaeus chinensis, and Pacific white shrimp, Litopenaeus vannamei, are reared. WSSV and HPV in pond seawaters ranged from $1.65{\times}10^3$ to $2.43{\times}10^9$ and from 0 to $4.43{\times}10^5$ copies/L of seawater, respectively. Of 20 ponds analyzed, all pond water and shrimp were positive for WSSv. L. vannamei showed higher susceptibility to WSSV than F chinensis. HPV was detected only in the pond water for F chinensis. In shrimp tissue, however, HPV was found in both species, with 23-times higher infection rate in F chinensis than L. vannamei. The total bacterial counts in the pond water ranged from $2.23{\times}l0^3$ to $1.98{\times}l0^5\;CFU/mL$. The variations in total bacterial count for each pond appeared to correlate to the variations of the WSSV load. Statistical analysis indicated that there was no significant difference (P>0.05) between the WSSV load in pond water and shrimp, and there was no relationship between total bacterial load and viral load in the pond water. However, a significant difference (P<0.01) was found between HPV load and L. vannamei and F chinensis pond water.

• Journal of Life Science
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• v.20 no.10
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• pp.1427-1432
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• 2010

This study was carried out to evaluate neutralization effects against WSSV using antiserum produced from recombinant envelop protein, rVP466 of WSSV. The VP466 gene of WSSV was cloned into pCold I expression vector and rVP466 was expressed in E. coli RIPL. The antiserum against rVP466 was produced in white rabbits (New Zealand white rabbit). The specific immunoreactivity to the antigen, rVP466, was confirmed by Western blot. The constant amounts of WSSV at $1{\times}10^4$ diluted stocks were mixed with various antiserum concentrations and then injected to the muscle of shrimp, Penaeus chinensis, for the neutralization challenge. The shrimps challenged with WSSV as a positive control and those with the mixture of WSSV and preimmune serum as a preimmune control showed 100% cumulative mortality at 17 days post challenge and 83% at 25 days post challenge, respectively. The shrimps challenged with 3 different mixtures of WSSV and rVP466 antiserum at ratios of 1:0.01, 1:0.1 and 1:1 showed 73%, 53% and 46% cumulative mortalities at 25 days post challenge, respectively. These results indicated that WSSV could be neutralized by the rVP466 antiserum. These results suggest that envelop protein VP466 is involved in the initial step of WSSV infection in shrimp.

• 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 Environmental Biology
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• v.35 no.4
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• pp.461-467
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• 2017

Shrimps infected with WSSV(White Spot Syndrome Virus) generally exhibit white spots in their inner space of carapaces as an acute clinical sign. In an effort to identify the correlation between this acute clinical sign and the condition, the index factors (RNA/DNA concentration and ratio, trypsin activity) were analyzed. A total 580 farmed Fenneropenaeus chinensis and 130 Lithopenaeus vannamei were collected from western and southern fifteen outdoor ponds in Korea. The status of the white spot pathology was divided into four stages (stage 0, stage I, stage II, and stage III), in accordance with the clinical signs as to the size and area of white spots. A significant decrease in RNA concentration and RNA/DNA ratio for multi-infected fleshy prawn (WSSV and vibrio sp.) occurred during the stage III (the whole carapace is covered with a white spot). In particular, RNA/DNA ratio was significantly lower as $1.47{\pm}0.04$ than other groups. A similar trend was also found in the single infection (WSSV), but the decrease was less than the multi-infection. In the species comparison, both species were vulnerable to the multi-infection, but L. vannamei was more sensitive than F. chinensis(ANOVA, p<0.05): A significant decrease in RNA concentration and RNA/DNA ratio was first found in stage II for the former species, while it was found in stage III for the latter species. Trypsin activity was also showed a similar tendency with nucleic acid variation. Multi-infected shrimp showed drastically decrease of trypsin activity. According to the results, clinical signs of the white spot under carapace have an only physiological effect on shrimp if they covered entirely with white spots.

• 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.

• 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.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.261-267
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• 2016

The present study examined the effect of the dietary administration of a halophilic bacterium Bacillus sp. Mk22 on the growth improvement of black tiger shrimp, Penaeus monodon, and reduced shrimp infection by Vibrio parahaemolyticus and white spot syndrome virus (WSSV). The shrimp were fed 45 days using three experimental diets: no addition (control), commercial probiotic, and Bacillus sp. Mk22. The shrimp treated with the halophilic bacterium Mk22 showed a significant improvement of growth (7.1 ± 0.21 g), survival (94.3 ± 0.58%), weight gain (178 ± 4.93 g), and reduced feed conversion rate (0.8 ± 0.03 g) compared with the shrimp in the other groups. The shrimp treated with Bacillus sp. Mk22 also showed a lower Vibrio count (0.02 ± 0.01 × 10² CFU/ml) in the shrimp culture water compared with the other groups. The shrimp in the three groups were challenged with either Vibrio or WSSV. For the Vibrio infection, no mortality was observed from water infection or oral feeding infection in the commercial probiotic group and Mk 22 group. For the WSSV infection, a 68% survival rate from water infection and 20% survival rate from oral feeding infection was observed on day 45 in the Mk22 group, while 100% mortalities were recorded at a much earlier time in both the control and commercial probiotic groups. The antioxidant enzyme activities, indicators of oxidative stress, such as catalase and superoxide dismutase, significantly decreased in both the Vibrio and WSSV-infected Mk22 groups compared with the other groups, indicating that Bacillus sp. Mk22 was effective in reducing oxidative stress, possibly due to the reduced infection.

• 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.

• Journal of fish pathology
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• v.24 no.2
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• pp.121-129
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• 2011

Aquatic animal raised in hatcheries play an important role in supplying seedling to stock enhancement and seed quality, especially, seed health is the key factor for survival in the field after release and for stocking effectiveness. We have inspected the hatchery-reared seeds of 33 marine species and 12 freshwater species for legally designated diseases in stock enhancement program in 2010. Results showed that abalone was the most abundant as 20.0% in the marine species group and then sea cucumber (15.6%), olive flounder (8.4%), rockfish (6.7%), black sea bream (6.3%) and swimming crab (6.1%) were followed. Crucian carp was the most abundant as 19.4%, and then eel (11.8%), Korean bullhead (10.9%), mandarin fish (10.8%), melanian snail (8.4%), catfish (7.7%) were followed in the freshwater species group. The total number of inspection cases for eight pathogens in this study were 2,105 and disqualification cases were 30 by detection of aquatic animals pathogens such as koi herpesvirus (KHV), red sea bream iridovirus (RSIV), white spot syndrome virus (WSSV) or viral haemorrhagic septicemia virus (VHSV).