• Journal of fish pathology
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• v.11 no.1
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• pp.43-50
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• 1998

In order to determine whether the tumor-inducing iridoviruses and the mortality-associated iridoviruses from cultured marine fish in Korea belong to same type, we compared the immunological characteristics of these viruses. The electrophoretical pattern of structural proteins of the tumor-inducing was different from that of the mortality-associated iridoviruses. The antigenicity of structural proteins of these viruses were identified by Western blotting using two monoclonal antibodies against tumor-inducing iridovirus. Two monoclonal antibodies recognized a 150 kDa structural protein of tumor-inducing iridoviruses showed. However, the structural proteins of the mortality-associated iridoviruses did not react with these monoclonal antibodies. These results demonstrate that the antigenicity of the structural proteins of tumor-inducing iridovirus is different from that of mortality-associated iridovirus, indicating that these two iridoviruses belong to different types.

• Archives of Pharmacal Research
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• v.22 no.5
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• pp.485-490
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• 1999

The mechanism of action of fish oil (FO), currently used in different chronic inflammatory conditions such as rheumatoid arthritis (RA), is not completely understood, although it is thought that it could alter the metabolism of endogenous autacoids. In addition, we hypothesized that the known capability of fatty acids (FA) of stabilizing serum albumin and perhaps other proteins, may be of pharmacological relevance considering that it is shared by other anti-rheumatic agents (e.g. nonsteroidal antiinflammatory drugs). Thus, we studied the effect of oral administration of FO and corn oil (CO), a vegetable oil with a different composition, on the stability of rat serum proteins, evaluated buy a classical in vitro method based on heat-induced protein denaturation. FO, and, to a lower extent, CO inhibited heat-induced denaturation of rat serum (RS): based on the inhibitory activity (EC50) of the major fatty acids against heat-induced denaturation of RS in vitro, it was possible to speculate the in vivo effects of palmitic acid (C16:0) and eicosapentaenoic acid (EPA, C20:5, n-3) may be more relevant than that of linolenic acid (C18:2). To better investigate this phenomenon, we extracted albumin from the serum of animals treated or not with FO with a one-step affinity chromatography technique, obtaining high purity rat serum albumin preparations (RSA-CTRL and RSA-FO), as judged by SDS-PAGE with Coomassie blue staining. When these RSA preparations were heated at $70^{\circ}C$ for 30 min, it was noted that RSA-FO was much more stable than RSA-CTRL, presumably due to higher number of long chain fatty acids (FA) such as palmitic acid or EPA. In conclusion, we provided evidences that oral administration of FO in the rat stabilizes serum albumin, due to an increase in the number of protein bound long chain fatty acids (e.g. palitic acid and EPA). We speculate that the stabilization of serum albumin and perhaps other proteins could prevent changes of antigenicity due to protein denaturation and glycosylation, which may trigger pathological autoimmune responses, suggesting that this action may be involved in the mode of action of FO in RA and other chronic inflammatory diseases.

• Journal of fish pathology
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• v.19 no.1
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• pp.45-54
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• 2006

The immunogenicity of Edwardsiella tarda was surveyed under two different culture conditions. In SDS-PAGE patterns of the outer membrane proteins (OMPs) extracts of E. tarda, grown under Trypic soy broth (TSB) and TSB supplemented iron chelate 2,2‘-dipyridyl iron-restricted condition, were examined. The results showed that the iron-regulated outer membrane protein (IROMPs) with molecular masses of 68 and 73 kDa were expressed by bacteria grown in iron-chelate TSB.The pathogenicity was examined by intraperitoneal injection with live E. tarda grown under TSB, iron-chelate TSB and iron-supplemented TSB. The result of pathogenicity test showed significantly high mortality in the group of live E. tarda grown under iron-chelate TSB.The effect of formalin killed cell (FKC) of TSB cultured bacteria and 2,2'-dipyridyl FKC (DP-FKC) of cultured bacteria on the iron-chelate TSB on the development of protective immunity in olive flounder was studied. The level of immune response was evaluated with immunized fish at 1, 2, 3 and 4 weeks after immunization. The numbers of specific antibody secreting cells (SASCs) showed significantly increased level at 2 week after immunization in each group. The agglutination titre of immunized fish was significantly high level at 3 weeks after immunization.The level of protection in olive flounder at 1, 2, 3 and 4 weeks after vaccination was examined by intraperitoneal challenge test with live E. tarda.

• Korean Journal of Ichthyology
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• v.18 no.4
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• pp.283-292
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• 2006

Expressed sequence tag (EST) analysis was conducted using a complementary DNA (cDNA) library made from the kidney mRNA of olive flounder (Paralichthys olivaceus). In the survey of 390 ESTs chosen from the kidney cDNA library, 250 ESTs showed significant homology to previously described genes while 140 ESTs were unidentified or novel. Comparative analysis of the 250 identified ESTs showed that 14 (5.6%) clones were representing 11 unique genes identified as homologous to the previously reported olive flounder ESTs, 198 (79.2%) clones representing 160 unique genes were identified as orthologs of known genes from other organisms, and orthologs were established for 38 (15.2%) clones representing 37 genes of known sequences with unknown functions. We also identified several kinds of immune associated proteins, indicating EST as a powerful method for identifying immunerelated genes of fish as well as identifying novel genes. Further studies using cDNA microarrays are needed to identify the differentially expressed transcripts after disease infection.

• Asian-Australasian Journal of Animal Sciences
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• v.4 no.3
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• pp.263-274
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• 1991

Digestibilities of nutrients and energy are among the most important parameters to be determined in feed evaluation research. The apparent digestibility coefficients (ADC) of protein, total carbohydrate (TCHO), fat, energy and amino acids were determined for 14 common feed ingredients using chromic oxide as external indicator with Israeli carp (Cyprinus carpio). The ingredients tested were; corn, corn starch, gelatinized starch, wheat middling, wheat grade inferior, corn gluten meal, rapeseed meal (solvent extracted), soybean meal (solvent extracted), blood meal (drum dried), feather meal(hydrolyzed), file fish meal (flame dried), sardine fish meal (steam dried), sardine fish meal (flame dried) and brewers yeast (dehydrated). The overall ADC values were high in Israeli carp showing high capacity to digest their feed ingredients irrespective of plant or animal sources. In addition the ADC of plant protein was high enough to support the successful supplementation of fish meal with other plant proteins.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.43 no.4
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• pp.307-313
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• 2010

Stanniocalcin 1 (STC1) is a glycoprotein hormone that is important in the maintenance of calcium and phosphate homeostasis in both fish and mammals. STC1 and its paralog STC2 are expressed in multiple tissues in fishes, although the physiological roles of piscine STCs are still unclear compared with those of mammals. In this study, we cloned olive flounder STC1 (ofSTC1) and ofSTC2 cDNAs into pET28a vector and used E. coli Rosetta (DE3) as the host strain for protein expression. Expression experiments were carried out using isopropyl-$\beta$-D-thiogalactoside (IPTG) and nickel affinity chromatography. We could identify the recombinant proteins as single 29.5 kDa (ofSTC1) and 33.2 kDa (ofSTC2) bands in the insoluble fraction on sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE). These results indicate that ofSTC1 and ofSTC2 were expressed as insoluble proteins in E. coli. Furthermore, the injection of ofSTC1 protein into juvenile tilapia resulted in a decrease of the serum calcium level. These results suggest that the purified fish STC1 and STC2 proteins may be used to elucidate the physiological role of STCs in fishes.

• 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.

• Korean Journal of Environmental Biology
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• v.26 no.3
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• pp.214-219
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• 2008

Changes in stress-associated biomolecules can be used as an important criterion for assessing the levels of environmental pollution because living organisms demonstrate contamination-stimulated stress responses. This study was conducted to determine the environmental status of Masan-Jinhae Bay, Korea, and its effects on marine organisms by investigating the endoplasmic reticulum (ER) dysfunction in the organs of the flat fish, Limanda yokohamae. ER dysfunction was evaluated via Western blot analysis of the ER stress proteins, immunoglobulin heavy chain binding protein (BiP) and C/EBP-homologous protein (CHOP), and the ER stress-associated protein caspase-12. The results showed that the amount of BiP and CHOP immunoreactivity in the flat fish from the bay area was much greater than that from the Gangneung, as a reference site. Similar to the ER stress proteins, the immunoreactivity of caspase-12 was also found to be elevated in the bay area when compared with that of Gangneung. These data suggest that the environmental status of Masan-Jinhae Bay induces the ER stress response, which is able to lead to phenotypic changes in marine organisms including fish.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.8
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• pp.1159-1165
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• 2016

The nutritional value of feed proteins and their utilization by livestock are related not only to the chemical composition but also to the structure of feed proteins, but few studies thus far have investigated the relationship between the structure of feed proteins and their solubility as well as digestibility in monogastric animals. To address this question we analyzed soybean meal, fish meal, corn distiller's dried grains with solubles, corn gluten meal, and feather meal by Fourier transform infrared (FTIR) spectroscopy to determine the protein molecular spectral band characteristics for amides I and II as well as ${\alpha}$-helices and ${\beta}$-sheets and their ratios. Protein solubility and in vitro digestibility were measured with the Kjeldahl method using 0.2% KOH solution and the pepsin-pancreatin two-step enzymatic method, respectively. We found that all measured spectral band intensities (height and area) of feed proteins were correlated with their the in vitro digestibility and solubility ($p{\leq}0.003$); moreover, the relatively quantitative amounts of ${\alpha}$-helices, random coils, and ${\alpha}$-helix to ${\beta}$-sheet ratio in protein secondary structures were positively correlated with protein in vitro digestibility and solubility ($p{\leq}0.004$). On the other hand, the percentage of ${\beta}$-sheet structures was negatively correlated with protein in vitro digestibility (p<0.001) and solubility (p = 0.002). These results demonstrate that the molecular structure characteristics of feed proteins are closely related to their in vitro digestibility at 28 h and solubility. Furthermore, the ${\alpha}$-helix-to-${\beta}$-sheet ratio can be used to predict the nutritional value of feed proteins.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.2
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• pp.100-105
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• 2006

This study was conducted to evaluate the effects of extruded pellet (EP) diets, as compared to a raw fish moist pellet (MP) diet for olive flounder, Paralichthys olivaceus, grown in commercial-scale aquaculture for 1 year. Four diets with duplication per diet were formulated for this experiment: two experimental EP diets (EP1 and EP2), one commercial EP diet (CEP), and a raw fish MP diet (MP). The MP diet consisted of 80% frozen horse mackerel and 20% commercial binder meal. Fish weighing $30.1{\pm}0.1 g$ ($mean{\pm}SD$) were distributed randomly to each aquarium as a group of 2,600 fish. Weight gain (WG) and feed efficiency ratio (FER) of fish fed EP2 and MP were higher (P<0.05) than those of fish fed CEP, while those of fish fed EP1 did not differ (P>0.05) from those fed EP2 and MP. However, fish fed the MP diet had a higher survival rate than fish fed the other diets. Fish fed EP2 had higher serum, phospholipids and total protein levels, and lower levels of serum glutamic-oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), and total cholesterol than fish fed MP (P<0.05). Dorsal muscle and liver proteins and lipid of fish fed EP1 were higher (P<0.05) than those of fish fed CEP, while those of fish fed EP1 and MP did not differ from those of fish fed EP2 (P>0.05). These results strongly suggest that EP1 could be developed to replace MP for grow-out stage production of olive flounder without adverse effects on growth performance.