• Journal of Life Science
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• v.19 no.9
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• pp.1328-1332
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• 2009

Anisakidosis is caused by anisakid nematodes (family Anisakidae) larvae which can cause not only direct tissue damage but also a severe allergic response related to excretory-secretion products. Lots of different species of anisakid larvae, including Anisakis simplex, Contracaecum, Goezia, Pseudoterranova, and Hysterothylacium, cause the anisakidosis. But it is difficult to diagnosis the species of larvae since the morphologies of larval anisakid nematodes are almost indistinguishable. In order to diagnosis the differential infections of larval anisakid nematodes, polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) of 18S rDNA - was conducted. Three major species of anisakid larvae including A. simplex, C.ontracaecum spp, and Goezia spp. were collected from mackerel (Scomber japonicus), mullet (Mugil cephalus), founder (Paralichthys olivaceus), eel (Astroconger myriaster) and red sea bream (Pagrus major). PCR amplified 18S rDNA from each species of anisakid larvae was digested with eight restriction enzymes including Taq I, Hinf I, Hha I, Alu I, Dde I, Hae III, Sau96 I, and Sau3A I. The original sizes of PCR amplified 18S rDNA were 2.0Kb in both anisakid larvaes and Goezia. Restrction enzymes including Hinf 1, Alu 1, Hha I, Dde 1 and Hae III cut differently and distinguished the A. simplex and Contracaecum type C'. However, Contracaecum type A showed two different restriction enzyme cutting patterns by Taq 1, Hinf I, Alu 1, and Dde 1. One of the patterns was the same as those of A. simplex, Contracaecum type C' and Goezia and the other was unique. These results suggest that PCR-RFLP pattern by Hinf 1, Alu 1, Hae I, Dde 1 and Hae III can be applied to differential diagnosis of human infection with A. simplex and Contracaecum type C'. Contracaecum type A needs further study of classification by morphological characteristics and genetic analysis.

• Journal of Life Science
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• v.21 no.8
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• pp.1149-1155
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• 2011

Myostatin (MSTN) belongs to the transforming growth factor-${\beta}$ superfamily or growth and differentiation factor 8 (GDF-8), and functions as a negative regulator of skeletal muscle development and growth. Previous studies in mammals have suggested that myostatin knock-out increased muscle mass and decreased fat content compared to those of the wide type. Recently, several studies on myostatin have beenconducted on the block myostatin signal pathway with myostatin antagonists and the MSTN regulation with RNAi to control myostatin function. This study was performed to analyze growth and muscle alteration of Oncorhychus masou by treatment with recombinant myostatin prodomains derived from fish. We designed myostatin prodomains derived from P. olivaceus (pMALc2x-poMSTNpro) and S. schlegeli (pMALc2x-sMSTNpro) in a pMALc2x expression vector, and then purified the recombinant proteins using affinity chromatography. The purified recombinant proteins were treated in O. masou through an immersion method. Recombinant protein treated groups did not show a significant difference in weight, protein, or lipid composition compared to the control. However, there was a difference in the average number and area for histological analyses in the muscle fiber. At twelve and twenty-two weeks from the initial treatment, there were differences in averagefiber number and area between the 0.05 mg/l treated-group and the control, but the numbers were similar to those of the control during the same time period. At twelve weeks, however, 0.2 mg/l treated-group had an increase in average fiber number and decrease in average fiber area compared to the control. At twenty-two weeks, the pMALc2x-sMSTNpro 0.2 mg/l treated-group was induced and showed a decrease in average fiber number and increase in average fiber area. The results between twelve and twenty-two weeks showed that the fiber numbers had decreased, whereas average fiberarea had increased due to sMSTNpro. It is understood that the sMSTNpro induced only hyperplasia at twelve weeks, after which it induced hypertrophy. Recombinant myostatin prodomains derived from fish may induce hyperplasia and hypertrophy in O. masou depending upon the time that has elapsed.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.1
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• pp.50-55
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• 1999

The proximate composition, mineral content, fatty acid composition, ATP related compounds, amino acid composition, color and texture were investigated with dorsal and ventral muscle from olive flounder, spotted flounder and hybrid (olive flounder m $\times$spotted flounder f). Spotted flounder and hybrid were higher in moisture content, and lower in crude protein content than those of olive flounder, Potassium content in hybrid was higher than that in olive flounder and spotted flounder. Hybrid was lower in calcium, iron, manganese content, and higher in magnesium content than olive flounder and spotted flounder. The contents of saturated fatty acid, unsaturated fatty acid and docosahexaenoic acid (DHA) in hybrid were intermediate level of spotted flounder and olive flounder. Fatty acid composition of dorsal muscle was slightly similar to ventral muscle. Adenosine triphosphate (ATP) and its related compounds contents and amino acid content in hybrid were intermediate level of spotted flounder and olive flounder, and these compounds of dorsal muscle were slightly similar to those of counterpart. The major amino acids such as aspartic acid, glutamic acid, leucine and Iysine were very similar to all the samples. Total amino acid and essential amino acid contents in dorsal muscle were slightly higher than those in ventral muscle. Free amino acid content and composition in hybrid were similar to spotted flounder, and free amino acid content in dorsal muscle was higher than in ventral muscle, The lightness of hybrid and spotted flounder was stronger than that of olive flounder, and was stronger in dorsal muscle. The breaking strength of hybrid was slightly lower than that of spotted flounder, and was stronger in dorsal muscle.

• Journal of Life Science
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• v.18 no.6
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• pp.789-795
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• 2008

To study the possible use of probiotics in fish farming, we evaluated antagonism of antibacterial strain Bacillus amyloliquefaciens H41 against the fish pathogenic bacterium Vibrio anguillarum NCMB1. The purification of growth inhibition factor produced by B. amyloliquefaciens H41 was achieved by obtaining supernatant of this bacterium. The growth inhibition factor was purified to homogeneity by 70% ammonium sulfate precipitation, DEAE-sephadex A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography, and sephadex G-50 gel filtration column chromatography with 40.8 fold of purification and 2.9% yield. The molecular weight of the purified growth inhibition factor was 48 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature for the growth inhibition factor were pH 7.5 and $30^{\circ}C$, respectively. The activity of growth inhibition factor was enhanced slightly by some metal ions, such as $Mg^{+2}$, $Mn^{+2}$, but was inhibited by the addition of $Co^{+2}$, $Hg^{+2}$, $Zn^{+2}$ and $Ag^{+2}$. NaCl stability of the growth inhibition factor was observed with 50% residual activity at 3% NaCl concentration. Toxicity test showed that the purified B. amyloliquefaciens H41 growth inhibition factor did not affect the live of Japanese flounder (Paralichthys olivaceus) and the effectiveness was 78% of residual lethality compared to commercial antibacterial agents.

• Korean Journal of Food Science and Technology
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• v.32 no.3
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• pp.550-563
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• 2000

This study was carried out in order to elucidate the fundamental data on the taste compounds between wild and cultured fishes produced on Chungmu and Wando at the southern coast areas of Korea. For this purpose, the food components of cultured fishes such as red sea bream Pagnus major, Sebastes pachycephalus and flounder, Paralichthys olivaceus being spot lighted for the main sea fish, the staple food and high economic fish were investigated and compared with those of the wild ones. There was a little appreciable difference in the proximate compositions of all the species from localities between wild and cultured fishes. But according to the growing conditions, wild fishes were higher in moisture contents and lower in crude lipid content than those of cultured fishes and little difference was seen in protein and ash contents between the two. With regard to the nucleotides and their related compounds, i.e. ADP, IMP and inosine were detected but ATP and hypoxanathine were not from them. On the other hand, there were little difference in the total taste compounds of all the species from localities and the growing conditions between wild and cultured fishes. But all the species were higher in IMP content. The total of seventeen amino acids were detected in samples. The highly contents of glutamic acid, lysine, aspartic acid, proline, leucine, alanine, valine and alginine were showed and less low contents of cystein, histidine, methionine, tyrosine and phenylalanine were detected. The total amino acids of the others were much alike in that composition. Little difference was seen from localities and the growing conditions between wild and cultured ones. The free amino acids were much alike in that composition of all the species. There was little difference in the free amino acid compositions all the species from localities and the growing conditions between wild and cultured fishes. But taurine was dominant, showing from 39% to 65% of the free amino acid content and it is followed by hydroxyproline, lysine, alanine and glycine in other. There were differences in the organic acid compositions of all the species from localities and the growing conditions between wild and cultured fishes. In addition, cultured fishes were more abundant in the total organic acid compositions than those of wild ones.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.115-121
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• 2002

To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.

• Journal of fish pathology
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• v.25 no.3
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• pp.211-219
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• 2012

The residue levels of ampicillin (AM) in cultured olive flounder, Paralichthys olivaceus (average 300g) at $20{\pm}1.0^{\circ}C$ were studied by oral, intramuscular and dipping administration (routes). The concentrations of AM in the plasma were determined by HPLC-UV detector. The average recoveries of AM in spiked samples between 0.01~10 ppm were ranging from 84.45% to 91.26% for plasma. The limit of detection for AM was 0.05 ppm by using this method. Plasma concentrations of AM were determined after oral dosage (10, 20 and 40 mg/kg body weight), intramuscular injection (5, 10 and 20 mg/kg body weight) and dipping (10, 20 and 40 ppm; 1 h). Samples were taken at 1, 3, 5, 10, 15, 24, 30, 48, 96, 144, 216, 264 and 360 h post-administration. In oral dosage of 10, 20 and 40 mg/kg, it's peak concentrations were $3.62{\pm}0.97$, $5.20{\pm}0.70$ and $11.18{\pm}0.87{\mu}g/ml$, respectively at 10 h post-administration, but AM was not measurable at 144, 360 and 360 h post-administration, respectively. In intramuscular injection of 5, 10 and 20 mg/kg, it's peak concentrations were $6.92{\pm}1.29{\mu}g/m1$, $9.89{\pm}2.22{\mu}g/ml$ and $19.85{\pm}2.97{\mu}g/ml$, respectively at 5 h post-administration, but AM was not measurable at 216, 264 and 264 h post-administration, respectively. In dipping of 10, 20 and 40 ppm, it's peak concentrations were $4.39{\pm}1.10$, $9.57{\pm}1.51$ and $11.61{\pm}1.92{\mu}g/ml$, respectively at 3 h post-administration, but AM was not measurable at 264, 264 and 360 h post-administration, respectively. Therefore, the plasma distribution and elimination levels of AM in olive flounder were dosage-dependant manner in all administration routes.

• Journal of fish pathology
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• v.25 no.3
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• pp.263-270
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• 2012

This study surveyed for the prevalence of parasites, bacteria and viruses in four fish species, olive flounder (Paralichthys olivaceus), red sea bream (Pagrus major), black sea bream (Acathopagrus schlegeli) and black rockfish (Sebastes schlegeli) in 2010. The survey was aimed to compare the pathogens detected from wild and cultured fish for an epidemiological study. Anisakis sp. was predominantly detected from wild olive flounder and red sea bream (58.6% and 41.7% respectively), but not from the cultured fishes, suggesting anisakid infection is rare in cultured fish. The wild fish get in contact with the anisakids through their prey such as small fishes or crustaceans which carry the anisakids; whereas the cultured fish are fed with formulated feed, free of anisakids. Bacterial detection rates from the wild fishes examined in the study were lower than those of cultured fishes. Vibrio sp. dominated among detected bacterial population in cultured olive flounder (18%). Since vibriosis is known as a secondary infection caused by other stressful factors such as parasitic infections, handling and chemical treatment, it seems that cultured olive flounder are exposed to stressful environment. Viruses diagnosed in the study showed difference in distribution between wild and cultured fishes; hirame rhabdovirus (HRV) (0.1%) and lymphocystis disease virus (LCDV) (3.9%) were detected in the cultured olive flounder, but not in the wild fish, and marine birnavirus (MBV) (1.7%) and red sea bream iridovirus (RSIV) (3.2%) were detected from the wild and cultured red sea bream, respectively. From the survey conducted, it can be concluded that even though some pathogens (Trichodina sp., Microcotyle sp., etc.) are detected from both the wild and cultured fish, pathogens such as Anisakis sp., Vibrio sp. and LCDV showed difference in distribution in the wild and cultured host of same fish species and this can be attributed to their environmental condition and feeding.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.4
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• pp.545-552
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• 1998

This study was undertaken to clarify the effects of electrical stimulation on the biochemical properties of plaice sarcoplasmic reticulum and myofibrils at early period of death. The plaices were electrically stimulated (110V/60 Hz) In sea water bath for 15, 35, and 60 seconds, and killed instantly by spiking at the head. Killed samples were investigated for the changes in $Ca^{2+}$-ATPase activity of FSR (fragmented sarcoplasmic reticulum), LSR (light SR), HSR (heavy SR), and SDS-PAGE pattern of FSR. $Ca^{2+}$-ATPase activity of FSR increased until $45^{\circ}C$ and inactivated over $50^{\circ}C$. $Ca^{2+}$-ATPase activity of FSR remarkably decreased according to the duration of electrical stimulation. Myofibrillar $Mg^{2+}$-ATPase activity of electrically stimulated plaices in the presence of $Ca^{2+}$ was higher than that of sample instantly killed by spiking. $Mg^{2+}$-ATPase activity of myofibrils increased by electrical stimulation and the activity decreased during storage at $5^{\circ}C$. Myofibrillar $Mg^{2+}$-ATPase activity in sample killed by spiking was not affected by $Ca^{2+}$ ion. Myofibrillar $Mg^{2+}$-ATPase activity of electrically stimulated sample in the absen-re of $Ca^{2+}$ decreased during storage at $5^{\circ}C$, whit $Mg^{2+}$-ATPase activity in unstimulated sample did not show any change. $Ca^{2+}$-sensitivity of myofibrils showed no differences between electrically stimulated sample and sample killed by spiking, and the was no change during at $5^{\circ}C$.

• Journal of fish pathology
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• v.3 no.1
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• pp.39-50
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• 1990

Chemical constituents in blood serum and electrolytes were mearsured to the cultured flounder, Paralichthys olivaceus classified by control group and establishments of mari-netcages from April 7, 1990 to May 8, 1990. The total protein, albumin and glucose were $5.74{\pm}0.83g/dl$, $0.94{\pm}0.22g/dl$ and $62{\pm}29mg/dl$, respectively in the blood serum constituents of control group. The potassium, sodium and calcium were $2.5{\pm}1.1\;mmol/l$, $169{\pm}17\;mmol/l$ and $5.0{\pm}1.7\;mg/l$, respectively in the electrolytes of control group. In the variation of chemical constituents and electrolytes in blood serum, glucose of PVC group was alone represented $66{\pm}19\;mg/dl$ as control level on the 9th after locomotion to mari-floating netcages, and was stable within period of experiment in comparision with the other group. Subsequently stable netron group was decreased $49{\pm}15\;mg/dl$ on the 12th. Total protein of netron group was alone represented $5.72{\pm}1.11\;g/dl$ as control level on the 27th. Albumin of PVC group was stable, and was retruned $1.00{\pm}0.18\;g/dl$ as control level on the 27th. Potassium of PVC and netron group were stable within priod of experiment. Sodium of PVC, pole-PVC group were stable, and were $169{\pm}17{\sim}178{\pm}10\;mmol/l$, $173{\pm}10{\sim}179{\pm}9\;mmol/l$ respectively in blood serum with in priod of experiment. According to the combined results of chemical constituents in blood serum and electrolytes classified by establishment of mari-floating netcages, the experiment group that were stabilized variation were in order of PVC, netron and pole-PVC group. But the other groups were extended the range of variation.