• Journal of fish pathology
• /
• v.16 no.1
• /
• pp.1-12
• /
• 2003

RT-PCR method was applied to detect and clone the nucleocapsid protein (N) gene and glycoprotein (G) gene for sequencing 5 Korean VHSV isolates from cultured olive flounder, Paralichthys olivaceus. Phylogenetic analysis was performed to investigate their relationship with the VHSV strains described previously and isolated from different geographical area. Generally, VHSV strains were separated phylogenetically according to the major geographical area of isolation: Genogroup I (American type), Genogroup Il (British Isles) and Genogroup ill (European type). This study revealed that all 5 Korean VHSV isolates were belonged to Genogroup I and closely related to Japanese Obama25 type.

• Journal of fish pathology
• /
• v.10 no.1
• /
• pp.21-29
• /
• 1997

We investigated on the prevalence and extermination of scuticociliatids parasitic on cultured japanese flounder, Paralichthys olivaceus in land-marine tank system of southern Korea from January to February in 1997. The gills and the skin showed the highest infection rate(60%), and the brain showed the lowest(22%). Also, fish secreted large quantity of mucus with a bleeding and ulcerated lesions on the infected sites. The number of the parasites in inflowing sea water, surface water and bottom water of farming tank ranged 0~1 individuals/$100m\ell$, 0~413 individuals/$100m\ell$ and $7\sim7.3{\times}10^4$ individuals/$100m\ell$, respectively. This parasite was died within 2 hours in 50~500 ppm, 48 hours of 10 ppm formalin or hydrogen peroxide, 1 hour in 50~500 ppm, 80 minutes of 10 ppm oligo chitosan and 10 minute in 100% but did not died until 48 hours in 10~70% fresh water.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
• /
• v.13 no.2
• /
• pp.140-146
• /
• 2008

Marine ecotoxicological standard method using fish larvae was established with the standard test species of Oryzias latipes(Japanese Medaka) and Paralichthys olivaceus(flounder) and with the 7 day $LC_{50}$ as endpoint. Test method referred to the USEPA(1994) with the replacement of test species found in the Korean water. Standard test species were selected in terms of the species supply and ecological importance in Korean waters. Japanese medaka can be reared with small tanks in the lab and has wide tolerance on salinity, and flounder eggs can be easily obtained from commercial fish hatcheries. General conditions for larval fish toxicity test are as follows. The possible salinity ranges for toxicity test were $0{\sim}35\;psu$ for medaka and >20 psu for flounder. Test type was designated as static non-renewal test if the dissolved oxygen in the test chamber does not fall below 4.0 mg/L. Ages of test species were selected as 7 days after hatched for medaka(about 5 mm TL) and 25 days for flounder(about 10 mm TL) because of the low natural mortality after these periods. Test can be accepted when the survival rates are over 80% in control. Also, species sensitivity on standard reference materials(copper, cadmium or zinc) must be provided with the toxicity test results.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.42 no.3
• /
• pp.204-208
• /
• 2009

The withdrawal time of erythromycin in cultured olive flounder was investigated to ensure the food safety of the fish treated with erythromycin. The oral administration of erythromycin was carried out using the experimental diet containing erythromycin (200 mg/kg) daily dose of 40 mg/kg body weight. The 45 day experimental period was broken into 7 days of habituation, 8 days of medication and 30 days of additional feeding without antibiotics. The erythromycin concentration in the flounder muscle had been increased gradually with medication. After 5 days of medication, the concentration increased to its maximum level of 6.05 mg/kg. After discontinuing the antibiotic, the erythromycin concentration decreased drastically and day 9 was below 0.1 mg/kg. The erythromycin concentration had slowly declined from the 6th to the 20th day after medication and disappeared completely after 25 days. From these results, the time needed to reduce the erythromycin level to the 0.2 mg/kg limit adopted by the ED and Japan was suspected to be 4-6 days. Therefore, a reasonable withdrawal time following ED and Japanese regulatory guidelines for erythromycin in the cultured flounder could be estimated to be 10 days.

• Asian-Australasian Journal of Animal Sciences
• /
• v.24 no.4
• /
• pp.463-470
• /
• 2011

Gonadotropin-releasing hormone receptor (GnRHR) gene is expressed at the anterior pituitary gland and plays a key role in gonad development. This study aimed to investigate molecular genetic characteristics of the GnRHR gene and elucidate the effects of single nucleotide polymorphisms (SNPs) of GnRHR gene on sex steroid level in Japanese flounder (Paralichthys olivaceus). We used polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and sequencing of the GnRHR gene in 75 individuals. We identified three SNPs in the GnRHR gene: P1 locus (C759A and C830T) in the coding region of exon2 which were both linked together and P2 locus (G984T) in the coding region of exon3, which added a new transcript factor (ADR1) and a new methylation site (CG). Only C830T of P1 leads to amino acid changes Thr266Ile. Statistical analysis showed that P1 was significantly associated with $17{\beta}$-estradiol ($E_2$) level (p<0.01) and gonadosomatic index (GSI) (p<0.05). Individuals with genotype BB of P1 had significantly higher serum $E_2$ levels (p<0.01) and GSI (p<0.05) than those of genotype AA or AB. Another SNP, P2, synonymous mutation, was significantly associated with GSI (p<0.05). Individuals with genotype AB of P2 had significantly higher GSI (p<0.05) than that of genotype AA. In addition, there was a significant association between one diplotype based on three SNPs and reproductive traits. The genetic effects for both serum $E_2$ level and GSI of diplotype D4 were super diplotypes (p<0.05). These results suggest that the SNPs in Japanese Flounder GnRHR are associated with $E_2$ level and GSI.

• BMB Reports
• /
• v.40 no.4
• /
• pp.459-466
• /
• 2007

The cDNA sequence of the Japanese flounder (Paralychthys olivaceus) IgD has been previously reported (GenBank accession no. AB052658) and this was followed by the detection of IgD mRNA expression in some flounder organ tissues. However, it has not been determined whether the flounder IgD gene is virtually expressed into IgD protein. To characterize the flounder immunoglobulins utilized in elucidating the mechanism, evolution and diversity of the flounder immune system, antibodies specific to IgD and IgM were necessary. In the present study, partial flounder recombinant IgD (rIgD), IgM (rIgM) and the conserved regions of IgD and IgM (rCIg) were produced by cloning the cDNA sequence using isotype specific primers which were designed to produce unique fragments of IgD and IgM specific amino acid sequences. The production of recombinant Igs was ascertained by SDS-gel electrophoresis and immunoblot analysis using anti-T7$\cdot}$Taq antibody. The produced recombinant Igs were purified using affinity columns, and used as immunogens. Antibodies specific to the isotype of flounder Igs were generated by immunizing rabbits with rfIgs and the antibodies produced were identified by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Specificities of the generated antibodies were evaluated by testing cross-reactivity between recombinant IgM and IgD. By ELISA, rabbit antibodies against the rfIgD fragment (anti-rfIgD) failed to recognize any kind of flounder serum Igs, whereas respective antibodies against rfCIg (anti-rfCIg) and rfIgM fragments (anti-rfIgM) reacted with serum Igs. Likewise, in immunoblot assays, though anti-rfIgD did not, both anti-rfCIg and anti-rfIgM bound with the ~85 kd flounder IgM heavy chain. By flow cytometry analysis, anti-rfCIg, anti-rfIgD and anti-rfIgM reacted with 6%, 3% and 6.5% of cells, respectively, suggesting that flounder IgD is not secreted in serum but expressed on flounder B-like cell surfaces as in mammals. Antibodies produced against recombinant flounder Igs could be used to develop sandwich assay systems for detecting flounder Igs and for further investigating the flounder immune system.

• Journal of fish pathology
• /
• v.11 no.2
• /
• pp.129-136
• /
• 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.

• Korean Journal of Ichthyology
• /
• v.19 no.3
• /
• pp.193-200
• /
• 2007

The samples of cultured flounder (Paralichtys olivaceus) were collected from Tongyeong and Yosu in Korea and Nagasaki in Japan and processed to analyses of body and serum. Body size and weight were not significantly different between Korea and Japan produced. But the body weight of average size of flounder produced from Nagasaki was the highest from the samples. The average of fish body was not significantly different from the samples. The water content of fish body was lower in samples from Nagasaki and fat content was higher in the samples from Nagasaki and followed by Yosu and Tongyeong. The percentage of body water, fat and protein contents was Nagasaki, Yosu and Tongyeong in the order of high values. Japanese flounder showed relatively high content of body fat and muscle. The cholesterol of serum was highest in the samples of Yosu. The triglyceride of serum was highest in the samples of Nagasaki. Lipoprotein was highest in the samples of Nagasaki and enriched fat content in LDL fraction.

• Journal of fish pathology
• /
• v.24 no.1
• /
• pp.1-9
• /
• 2011

Viral haemorrhagic septicaemia virus (VHSV) is an epidemic virus in olive flounder Paralichthys olivaceus farms in Korea, since the virus have first isolated in 2001. In the present study, partial glycoprotein (G) gene nucleotide sequences of seven Korean VHSV from cultured olive flounder and wild marine fishes in coastal areas of Korea were analyzed to evaluate their genetic relatedness to worldwide isolates. Phylogenetically, all Korean VHSV formed only one minor cluster including Japanese isolates, in genotype IVa, while the North America isolates formed a different minor cluster in genotype IVa. These results suggest that Korean VHSV could be an indigenous virus in Korean and Japanese coastal areas, but have not been introduced from North America.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.35 no.3
• /
• pp.237-241
• /
• 2002

This study was performed both to explore the host of nervous necrosis virus (NNV) between mariculturing fish species and to examine the phylogenic position of the NNV in Korea. NNV was confirmed on the basis of histopathological and molecular biological examination, then VNN infection was Preyed from either moribund or dead fishes including red drum, Sciaenops ocellatus; oblong rock fish, Sebastes oblongus and flounder, Paralichthys olivaceus. As a result of sequencing for a part of ms, virus from red drum was showed $98\%, $97\%, $86\% and $74\% homology with oblong rock fish, grouper, Japanese flounder and striped jack, respectively. On the other hand, NNV from oblong rock fish was demonstrated $96\%, $85\% and $72\% homology with grouper, Japanese flounder and striped jack, respectively. NNV from red drum and oblong rock fish was exhibited phylogenically distant from the representative NNV, SJNNV originated from striped jack. On the contrary, the viruses appeared to be similar species with Taiwan NNV isolated from culturing grouper.