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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal of fish pathology
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Journal DOI :
The Korean Society of Fish Pathology
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Volume & Issues
Volume 6, Issue 2 - Dec 1993
Volume 6, Issue 1 - May 1993
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Studies on Viral Disease of masu salmon, Oncorhynchus masou-II Isolation of infectious hematopoietic necrosis virus form masu salmon fry
Sohn, Sang-Gyu ; Park, Myoung-Ae ; Park, Jeong-Woo ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 87~92
In February of 1990, an epizootic disease to masu salmon. Onchorynchus masou cultured at the hatchery of trout in Samchuk. Kwangwondo have broken out and induced heavy mortality. An infectious hematopoietic necrosis virus(IHNV) was isolated from diseased masu salmon fry by the use of fish cell line, CHSE-214. This IHNV isolated from masu salmon was compared with USA isolates of IHNV, SRCV and RB-76 by analysis of virion proteins in sodium dodecyl sulfate poly-acrylamide gel electrophoresis (SDS-PAGE) and neutralization tests with two monoclonal antibodies raised against SRCV(MAb SRCV/A4) and RB-76(MAb RB/B5). In the antigenicity and the size of structural proteins. this IHNV, SCS atrain was smilar to RB-76 belonged to the electropherotype I proposed by Hsu et al.(1986).
Characterization of the cloned RNA1 gene of Saccharomyces cerevisiae
Song, Young-Hwan ; Kim, Dae-Young ; Kim, Jin-Kyung ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 93~101
The RNAI mutation of Saccharomyces cerevisia is a recessive and temperature sensitive lethal mutation which interferes with the production of mRNA, rRNA, and tRNA. However, the precise role of RNAI gene have not been revealed until yet. We have cloned rna1-1 mutant gene from rna1-1 mutant yeast strain(R49 ; trpl, ura3-52, rna1-1). The 3.4kb BglII fragment of wild type RNAI clone(81-2-6) contains whole RNAI gene. The genomic southern blotting with BglII digested R49 genomic DNA as a probe shows the unique and identical band with wild type 3.4kb BglII fragment. Therefore, We prepared partial BglII genomic library(3~4kb BglII fragments) into BamH I site of pUC19. The rna 1-1 mutant clone was screened with Digoxigenin(DIG)-lableled probe by high density colony hybridization. The 5'-flanking region of rna1-1 gene was sequenced by dideoxy chain termination method. The 5'-flanking sequence of RNAI gene contains three TATA-like sequence ; TAATA, TATA and TTTTAA at position of -67, -45, and -36 from first ATG codon respectively. The 5'-flanking region of wild type RNA I gene from ATG codon to -103nt was deleted with Bal31 exonuclease digestion, generating
/RNA I. After constructing
I (consists of -103nt deleting RNA I gene, URA3 gene,
rep. origin), pYEPrna1-1(consists of Xba I fragment of pUCrna1-1. URA3 gene,
rep. origin), and pYEPRNAI. each plasmid was transformed into host strain(trpl, ura3-52, rna1-1) by electroporation, respectively. Yeast transformant carrying
I did not complement the thermal sensitivity of rna1-1 gene. It means that TATA-like sequences in 5'-flanking region is not TATA sequence for transcribing RNAI gene and there may be other essential sequence in upstream region for the transcription of RNAI gene.
Molecular Cloning of Chitinase Genes Family from Serratia marcescens
Song, Young-Hwan ; Kweon, Oh-Gun ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 103~110
Total genomic DNA library of Serratia marcescens was prepared by inserting Sau3AI partial digesting fragments(above 5 kb) into the dephosphorylated BamHl site of pUC19. In primary screening, two colonies were selected by observing the halo around E. coli transformants grown on the swollen colloidal chitin media. Secondary screening was performed by soaking two colonies with a few drops of 4-methylumbelleliferryl N-acetyl-
-D-glucocosaminide(4-MuNGlcNAc). As 4-MuNGlcNAc is a specific, fluorogenic substrate for chitinase, the positive clones produce light fluorescence by the exposure under the long wave U.V. light(360 nm). From genomic DNA library derived from pUC19, we have isolated two different chitinase clones, pCH1(11.0Kb) and pCH2(7.5Kb), which show completely different restriction map to each other. The cross-hybridization of pCH1EA and pCH2 have not revealed any hybridization signals to each other.
Gene Expression and its Regulation on Several Different B Cell Population by using in situ Hybridization technique
Jeong, Hyun-Do ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 111~122
The mechanism by which
region gene segments is selected in B lymphocyte is not known. Moreover, evidence for both random and nonrandom expression of
genes in matured B cells has been presented previously. In this report, the technique of in situ hybridization allowed us to analyze expressed
gene families in normal B lymphocyte at the single cell level. The analysis of normal B cells in this study eliminated any posssible bias resulting from transformation protocols used previously and minimized limitation associated with sampling size. Therefore, an accurate measure of the functional and expressed
gene repertoire in B lymphocyte could be made. One of the most important controls for the optimization of in situ hybridization is to establish probe concentration and washing stringency due to the degree of nucleotide sequence similarlity between different families which in some cases can be as high as 70%. When the radioactive
RNA probes are tested on LPS-stimulated adult spleen cells,
/slide shows low background and reasonable frequency of specific positive cells. For the washing condition. 40~50% formamide at
is found to be optimum for the
probes. The analyzed results clearly demonstrate that the level of each different
gene family expression is dependent upon the complexity or size of that family. These findings are also extended to the level of
gene family expression in separated bone marrow B cells depend upon the various stage of differentiation and conclude no preferential utilization of specific
gene family. Thus, the utilization of VH gene segments in B lymphocyte of adult BALB/c mice is random and is not regulated or changed during the differentiation of B cells.
Changes in Blood Parameters of the Cultured Flounder Paralichthys olivaceus Artificially Infected with Staphylococcus epidermidis
Sim, Doo-Saing ; Jung, Sung-Hee ; Park, Hyung-Sook ; Chun, She-Kyu ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 123~131
The cultured flounder(Paralichthys olivaceus) was injected with Staphylococcus epidermidis, various hematological and blood chemical changes were monitored over 96 hours. Red blood cell count, hemoglobin hematocrit. mean corpuscular hemoglobin concentration and mean corpuscular hemoglobin were significantly depressed after 24 to 48hours. Total protein, albumin, globulin and total cholestrol were significantly increased by the 24 or 48 hours, Glucose, bilirubin and transaminase were significantly depressed by 24 to 48hours. Erythrocytes were gotten shorter with round-shaped after 48hours inoculated with S. cpidermidis. Hemolytic erythrocytes and neutrophils were showed after 72hours inoculated with S. epidermidis. Price-Jones curve was transformed for left shift after 48hours inoculated with S. epidermidis, therfore staphylococcia appeared hemolytic anemia in the artificially infection.
ENZYMATIC STUDIES ON VITAMIN B6 METABOLISM
Kim, Young-Tae ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 133~142
Vitamin B6(pyridoxine, pyridoxamine. and pyridoxal) is a dietary requirement in relatively small quantities for growth, health, and function in animals and fish. The metabolically active B6 is pyridoxal-5-phosphate(PLP). It does function as a coenzyme in number of enzymes(PLP-dependent enzymes) in which amino acids are metabolized, including decarboxylases, aminotransferases, sulfhydrases, tryptophanase, and hydroxylases. Vitamin B6 requirement is higher for fish because fish are fed much higher protein diet than land animals. B6 is also involved in metabolism of carbohydrates and lipids and essential for the synthesis of heme and serotonin. Deficiency signs in fish develop quickly, in cluding nervous disorders, convulsions, poor swimming coordination, skin lesions, edema, exophthalmos, and tetany. The conversion of vitamin B6 to metabolically active form(PLP) is catalyzed by pyridoxal kinase and pridoxine(pyridoxamine) oxidase. In this review, we summarized in detail the enzymatic studies on vitamin B6 metabolism and about the mechanisms and properties of a PLP-dependent enzyme.
The Development Ceroidosis in Cultured Flounder, Paralichthys olivaceus
Lee, Chang-Hoon ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 143~161
1. For the induction of ceroidosis, the flounders were fed on the different Peroxidation Value(POV) diets for 6 months from April 1 to September 27, 1991. The growth of the flounder fed on the high POV diets, 90.4mEq/kg and 36.9mEq/kg, were markedly decreased after 3 months and 4 months respectively, compared to those fed on the two different low POV diets, 15.5mEq/kg or 15.2mEq/kg. 2. Histologically, it was hard to observe any pathological signs in the tissues of the control group with normal diets(POV 10.5mEq/kg). With the increasing of POV values of diets, severe ceroid accumulation and parenchymal degenerative lesions were found in the liver, spleen and kindeny. However, any abmormalities were not observed in muscle and intestine been in high POV diets fed flounder.
The treatment of Ceroidosis in Cultured flounder, Paralichthys Olivaceces
Lee, Chang-Hoon ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 163~175
1. By feeding on high Peroxidation Value(POV : 90.4mEq/kg) diets for 180 days, the cultured flounder were induced ceroidosis antificially. 2. The supplementation of glutathione to the diets having POV of 15.5mEq/kg improved the growth of the flounder that had shown depressed by the previous high POV diets(90.4mEq/kg). However, we found very much the same degree of improvement on growth by the supplemented diets of glutathione from 10mg/kg/day to 40mg/kg/day. 3. After 7 days, the flounder fed on the diets supplemented with 10, 30mg/kg/day glufthione start the recover from the accumulated ceroid and damaged tissues. These treated flounders with glutathione showed completely normal histlogical signo after 21 days. The effect of glutathione treatment with high concentration(30mg/kg/day) was better than that of low concentration(10mg/kg/day).
Prophylaxis of Ceroidosis in Cultured Flounder
Lee, Chang-Hoon ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 177~189
1. For the prophylaxis of ceroidosis Vitamin E and C supplemented diet(POV 90.4mEg/kg) was used for flounder culture during 80 days. 2. The supplementation of Vitamin E and C with our tested concentrations on the high POV diet supported almost normal growth of flounder. It is very much comparable with the abnormal growth of flounder fed the diet of high POV. 3. The best prophylactic effect were appeared the high POV diet supplemented with Vitamin E 1mg/g diet and Vitamin C 2IU/g diet. These results were analyzed by histological observation of tissues.
Viral diseases of Japanese flounder(Paralichthys olivaceus) in Japan
Nakai, Toshihiro ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 191~195
With the rapid progress in seed production techniques, aquaculture production of economically important species of marine fish has been accelerated in Japan. Howecer, mass mortalities due to viral infections as well as other microbial infections have often occurred during the seed production and grow-out stages. Among these diseases, four viral diseases have been known in cultured Japanese flounder (Paralichthys olivaceus) since around 1980. In this paper, viral diseases of cultured flounder in Japan are briefly reviewed, with special attention to two viral diseases. viral epidermal hyperplasia and rhabdovirus infection which are relatively important because of their frequent occurrence. Viral epidermal hyperplasia is characterized by fin opacity and associated with high mortality in larval flounder Electron microscopy of affected epidermal cells and transmission experiments with tissue filtrates demonstrated that the disease was caused by a herpesvirus but the agent has not been isolated in fish cell lines. On the other hand, rhabdovires infection occurrs in juvenile and production size fish with hemorrhage in the skeltal muscle and fins, congestion of the gonads, and ascites. A rhabdovirys was isolated in RTG-2 cells from the diseased flounder as a causative agent, which was designated hirame rhabdovirus (HRV) or Rahbdovirus olivaceus. HRV is serologically distinguishable from other known fish rhabdoviruses. Intensive researches on these viral diseases started in 1980th. but properties of the causative agents and infection mechanisms have not been fully investigated. This results in difficulty in controlling these diseases.
Bacterial diseases of flounder, Paralichthys olivaceus
Kanai, Kinya ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 197~204
Flounder culture has been developed mainly in the western parts of japan, and, to date, following six bacterial diseases have been reported. Bacterial white enteritis occurs in 16 to 30-day-old flounder larvae and often causes mass mortality in seed production. Bacterium named Vibrio sp. INFL invades and multiplies in the mucosae of posterier part of intestine, and causes desquamative enteritis. Gliding bacterial disease occurs mostly in juvenile stage and in spring to summer. Diseased signs are partial discoloration and erosion of skin and fins. Histologically, epidermis are removed, and the causative bacterium, Flexibacter maritimus, multiplies on the surface of demis and invades into the muscular tissue. Vibriosis caused by Vibrio anguillarum and related organisum is one of the well-known diseases among marine fish. Outbreaks of the disease in flounder culture are relatively few, but mass mortalities in fingerlings due to the disease were reported. An outbreak of nocardiosis in the autumn of 1984 has been reported, but since then the disease scarcely occurred. The disease is characterized by formation of abscesses under the skin and white nodes in the gill, heart, spleen and kidney. Streptococcicosis occurs frequently in recent years. Beta-hemolytic streptococcus is the causative bacterium, which possesses the same biochemical and serological characteristics as
-streptococci isolated from some marine and freshwater fish, and is seemed to related to Streptococcus iniae. Edwardsiellosis is the disease that causes most damage in flounder culture in Japan. Characteristic symptoms are swelling of abdomen and intestinal protrusion from the anus due to accumulation of ascites. Edwardsiella tarda, a well-known pathogen of freshwater fish, is the causative bacterium of the disease.
광어(Paralichtys olivaceus)의 스쿠치카감염증(感染症) -스쿠치카섬모충(纖毛蟲)의 배양성상(培養性状).약제감수성(藥劑感受性).병원성(病源性)-
Journal of fish pathology, volume 6, issue 2, 1993, Pages 205~218
On the development of hirame(Paratichtys olivaceus) culture, outbreak of scuticociliata infection was reported to cause severe damage in Japan. To establish effective measures for isolation and cultivation of this ciliate, we tried to culture this pathogenic ciliate using medium for bacteria and fish cell lines in vitro. Scuticociliata from the brain tissues of infected fish was aseptically inoculated to CHSE-214 cells cultured in MEM-10 without antibiotic. Scuticociliata grew well and the number of ciliate reached
at temperatures of
for 10d. The number of ciliate cultured in the cell lines is 10 times higher than the numbers cultured in the liquid medium alone. This ciliata could be cloned by dilution method. Scuticociliata isolated could grow well on 42 different cell lines that were established from marine fish, warm freshwater fish, and salmonids. This ciliate could be preserved in liquid nitrogen for more than 6 months. Subsequently, we observed the optimal temperature and salinity for growth, and tested the sensitivities of this organism to formaldehyde, flagyl(Metronidazole), Ekuteshin(Combination compound of sulfamonometoxin and ormethoprim), and ozonixation. Optimal temperature for growth was
and salinity was 1.0 to 1.5%. Washed scuticociliata was killed by formaldehyde at the concentration of 50ppm for 10min, but was not completely killed even at a high concentration of 400ppm for 20min in MEM-5. Flagyl and Ekuteshin can inhibit the growth of scuticociliata at the concentration of 1,000 and
in MEM-10, respectively. More than 99% of this scuticociliata could be killed by ozonization at a dose equivalent to
oxidant for 30sec in sea water. Isolated scuticociliata showed the pathogenicity to the cultured hirame by artificial infection(I. P. injection,
/fish). The number of scuticociliata in the water could be counted by most probable number(MPN) method using tissue culture, and the minimum detectable number was
. The number in the reservoir tank for water supply to the culture tank was 110 cells/l. After cleaning by elimination of the sediments from of the reservoir tank and disinfected with formaldehyde, number of scuticociliata decreased and was counted less than
and infection rate of cultured hirame was decreased.
Control Methods of Diseased of Japanese Flounder, Paralichthys olivaceus, used in Fish Farms, in Japan
Mizuno, Yoshitsugu ;
Journal of fish pathology, volume 6, issue 2, 1993, Pages 219~231
The author introduces the preventive and therapeutic methods of diseases in Japanese flounder which have been conducted by the Fish Disease Laboratory, the Mikamewan Fishery Cooperative Union, Ehime Prefecture, since 1982. Prevention 1. Addition of a sand substrate in the culture pond was effective to prevent ulcer disease. 2. Bathing in 0.5ppm of copper ion was effective to prevent some parasites. 3. Low stocking density of the fish reduced an incidence of edwardsiellosis or gliding bacterial disease. 4. Removal of the diseased fish prevented thd spread of lyphoeystis. 5. So-called
-water was effective to prevent the fry from some diseases. 6. Immersion of the juvenile in a sodium nifrusylate solution during transportation was effective to prevent gliding bacterial disease. Therapy 1. Sodium nifrustylate or oxytetracycline was effective to cure gliding bacterial disease. 2. Bathing in formalin(150ppm) or freshwater was effective to cure scuticociliatidosis. 3. Erythromycin was effective to cure
-hemolytic Streptococcus sp. infection. 4. Bathing in a hydrogen peroxide solution(1.5%) was effective to cure white spot disease.