• Korean Journal of Fisheries and Aquatic Sciences
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• v.45 no.2
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• pp.154-160
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• 2012

This study the effects of Cu exposure on biochemical factors in the hemolymph and hepatopancreas of the abalone $Haliotis$ $discus$ $hannai$. Abalone were exposed to 0, 5, 10, 20 and 40 ${\mu}g/L$ Cu for 4 weeks. The calcium concentrations in hemolymph were decreased significantly on exposure to 20 and 40 ${\mu}g/L$ Cu after 2 weeks. The aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities in abalone hemolymph were markedly elevated after exposure to 40 ${\mu}g/L$ Cu for 4 weeks. The hepatopancreas superoxide dismutase (SOD) and catalase (CAT) activities were also significantly increased by exposure to 20 ${\mu}g/L$ Cu for 4 weeks. These biochemical factors may represent a convenient method of monitoring heavy metal pollution in coastal areas. From these results, we conclude that a high copper concentration (40 ${\mu}g/L$) in water may curtail hemolymph homeostasis and anti-oxidative reactions in abalone.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.4
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• pp.388-395
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• 2017

We investigated the toxicity of zinc and aluminum hot-dip galvanized sheet steel to abalone Haliotis discus hannai via changes in the gill and hepatopancreas using histological and transmission electron microscopy analysis. Experimental groups were composed of one control and four exposure conditions (direct or indirect exposure to zinc and aluminum hot-dip galvanized sheet steel). In the control group, aluminum exposure groups (direct and indirect), and indirect zinc exposure group, abalone mortality was not observed until the end of the experiment, and no histopathological changes were observed in the gill and hepatopancreas. However, the direct zinc exposure group exhibited 100% mortality. Ultrastructural analysis of the cytoplasm of ciliated and microvilli-bearing epithelial cells from gill filaments revealed electron-dense vesicles near the cell membrane and disruption of the nuclear membrane. We also observed swollen mitochondria and a loss of mitochondrial cristae. The hepatopancreas showed similar changes, and we detected highly electron-dense particles within the vesicles. These results suggest that abalone exposed directly to zinc hot-dip galvanized sheet steel experience acute toxicity, causing damage to cell organelles in the gill and hepatopancreas and, finally, inducing mortality.

• Fisheries and Aquatic Sciences
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• v.3 no.3_4
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• pp.188-194
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• 2000

A protease purified from hepatopancreas of shrimp, Penaeus japonicus, had maximum activity at $70^{\circ}C$ and in neutral and alkaline pH ranges. Specific activity at optimum reaction condition of the protease was estimated to be approximately 12 U/mg/min. The protease was stable in neutral and alkaline pH ranges and activity was retained after heat treatment at $50^{\circ}C$ for 30 min. Apparent $K_m$ and $V_{max}$ value against casein substrate were estimated to be $0.29\%$ and $7.8see^{-1}$, respectively, and those against N-CBZ-L-tyrosine p-nitropheny1 ester (CBZ­Tyr-NE) were 0.38 mM and $2,400 see^{-1}$, respectively. The N-termina1 sequence of the protease showed high homology to the trypsin from same species and the proteases from shrimp. Myosin heavy chain (MHC) from shrimp tail meat was the most susceptible to the protease and actin/tropomyosin were degraded progressively during 4 hr incubation, but to a lesser degree than MHC.

• The Korean Journal of Food And Nutrition
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• v.19 no.4
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• pp.466-472
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• 2006

In order to develop materials of crab-like flavorant, taste compounds including physicochemical characteristics were analyzed in red snow crab cooker effluent(RSCCE) and hepatopancreas. The $30\;^{\circ}Brix$ was a suitable condition in from 1.5 to $40\;^{\circ}Brix$ RSCCE by sensory evaluation. Lactic acid and succinic acid were major compounds in non-volatile organic acids detected in both $30\;^{\circ}Brix$ RSCCE and hepatopanceras. The 5 compounds such as AMP, HxR, IMP, ATP and GMP were major in ATP related compounds of $30\;^{\circ}Brix$ RSCCE, whereas 3 compounds including IMP, GMP and Hx in hepatopanceras. The content of total free amino acids in hepatopancreas was 5.6 times higher than in $30\;^{\circ}Brix$ RSCCE. The major compounds in $30\;^{\circ}Brix$ RSCCE were followed by methionine, lysine, arginine, valine, histidine, alanine, hydroxy proline, and glycine in that order, whereas methyl histidine, leucine, alanine, glutamic acid, glycine, valine, threonine, taurine, isoleucine, and serine were followed in hepatopancreas. By adding 0.5%(w/w) hepatopancreas in $30\;^{\circ}Brix$ RSCCE, crab meat-like odor was kept high level by sensory evaluation.

• Fisheries and Aquatic Sciences
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• v.19 no.4
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• pp.21.1-21.11
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• 2016

Background: The evaluation of suitable reference genes as normalization controls is a prerequisite requirement for launching quantitative reverse transcription-PCR (RT-qPCR)-based expression study. In order to select the stable reference genes in abalone Haliotis discus hannai tissues (gill and hepatopancreas) under heavy metal exposure conditions (Cu, Zn, and Cd), 12 potential candidate housekeeping genes were subjected to expression stability based on the comprehensive ranking while integrating four different statistical algorithms (geNorm, NormFinder, BestKeeper, and ${\Delta}CT$ method). Results: Expression stability in the gill subset was determined as RPL7 > RPL8 > ACTB > RPL3 > PPIB > RPL7A > EF1A > RPL4 > GAPDH > RPL5 > UBE2 > B-TU. On the other hand, the ranking in the subset for hepatopancreas was RPL7 > RPL3 > RPL8 > ACTB > RPL4 > EF1A > RPL5 > RPL7A > B-TU > UBE2 > PPIB > GAPDH. The pairwise variation assessed by the geNorm program indicates that two reference genes could be sufficient for accurate normalization in both gill and hepatopancreas subsets. Overall, both gill and hepatopancreas subsets recommended ribosomal protein genes (particularly RPL7) as stable references, whereas traditional housekeepers such as ${\beta}-tubulin$ (B-TU) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were ranked as unstable genes. The validation of reference gene selection was confirmed with the quantitative assay of MT transcripts. Conclusions: The present analysis showed the importance of validating reference genes with multiple algorithmic approaches to select genes that are truly stable. Our results indicate that expression stability of a given reference gene could not always have consensus across tissue types. The data from this study could be a good guide for the future design of RT-qPCR studies with respect to metal regulation/detoxification and other related physiologies in this abalone species.

• Journal of fish pathology
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• v.14 no.2
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• pp.81-90
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• 2001

The histological responses of the flounder, Paralichthys olivaceus to copper were examined in the gill, hepatopancreas and kidney. In control group, from 5 weeks mucous cells and chloride cells were increased in the gill, and numerous hemocytes and some melano-macrophagocytes were observed in the hepatopancreas and kidney. The minimum concentration for histological responses was 0.05mg/$\ell$/7d. In this group gill and hepatopancreas showed chloride cell activation, hepatocyte activation, pancreatic zymogen reduction, and congestion, and melanomacrophagocytes were observed in the kidney. From the histological observations, the critical concentrations for dysfunctionality were 0.18mg/$\ell$/21d in the gill, 0.18mg/$\ell$/14d in the hepatopancreas and 0.08mg/$\ell$/14d in the kidney, respectively.

• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.201-208
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• 1998

A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

• Fisheries and Aquatic Sciences
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• v.2 no.2
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• pp.218-225
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• 1999

A protease without tryptic and chymotryptic activities was purified from the hepatopancreas of shrimp, Penaeus orientalis, using Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, Mono-Q, and gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 27kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS­PAGE). The amino acid composition of the protease was different from that of protease from P. japonicus or trypsin from P. orientalis. The protease was completely inhibited by benzamidine, $N\alpha-p-tosyl-L-lysine$ chloromethyl ketone (TLCK), and phenylmethylsulfonyl fluoride (PMSF) and was not affected by leupeptin, pepstatin, N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), iodoacetate, and ethylenediamine tetra acetate (EDTA). The enzyme did not have any activity against Na-benzoyl-DL-arginine p-nitroanilide (BAPNA) or N-benzoyl-L-tyrosine ethyl ester (BTEE) which are specific substrates of trypsin and chymotrypsin, respectively. However, the protease showed hydrolytic activity for a carboxyl terminal of Tyr, Trp, Phe, Glu, and Cys.

• Fisheries and Aquatic Sciences
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• v.17 no.2
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• pp.181-187
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• 2014

This study was performed to identify the optimum fractionation method and conditions to obtain exopeptidase-active fractions from octopus hepatopancreas (HP) crude extracts (CEs) using four techniques: solid ammonium sulfate fractionation, polyethylene glycol (PEG) fractionation, anion exchange chromatography, and gel filtration chromatography. The fractions with the highest total activity toward L-leucine-p-nitroanilide (Leu-pNA) were fraction IV from the ammonium sulfate and PEG fractionation, and fraction II in ion exchange and gel filtration chromatography. The total exoprotease activity of these fractions was highest in fraction IV (4,050.20 U) of ammonium sulfate fractionation, followed by fraction II (3,600.28 U) from gel filtration chromatography, fraction IV (2,861.30 U) from PEG fractionation, and fraction II (2,576.28 U) from ion exchange chromatography. These results suggest that ammonium sulfate fractionation using 60-80% ammonium sulfate was the most efficient method for separating the exoprotease active fractions from CEs of octopus HP.

• Journal of fish pathology
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• v.25 no.2
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• pp.103-109
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• 2012

The objective of this study was to investigate the effects of exposured $Cu^{2+}$ on growth and bioaccumulation of abalone Haliotis discus hannai. Abalone were exposed to various concentration of $Cu^{2+}$ (0, 5, 10, 20, $40{\mu}g/L$). Bioaccumulation of tissues, hepatopancreas, muscles and gills were measured. hepatopancreas and gills $Cu^{2+}$ concentration of abalone increases to extent during the 4 weeks accumulation time. But muscles showed no significant changes, with respect to control. These data indicate that abalone Haliotis discus hannai hepatopancreas and gills can be considered adequate target tissues for waterborne exposured of $Cu^{2+}$. Weight growth rate of abalone exposed to $Cu^{2+}$ was significantly decreased in 20 and $40{\mu}g/L$ groups compared to control. This study revealed that high $Cu^{2+}$ concentration (${\geq}20{\mu}g/{\ell}$) reduced growth of abalone. These data indicate that excessive waterborne $Cu^{2+}$ can affect the toxicity of xenbiotics to abalone through alterations in growth rate. Thus, environmental standard of $Cu^{2+}$ $20{\mu}g/L$, should be considered a potential source of variation in toxicological studies with abalone.