Go to the main menu
Skip to content
Go to bottom
REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
> Journal Vol & Issue
Journal of fish pathology
Journal Basic Information
Journal DOI :
The Korean Society of Fish Pathology
Editor in Chief :
Volume & Issues
Volume 22, Issue 3 - Dec 2009
Volume 22, Issue 2 - Aug 2009
Volume 22, Issue 1 - Apr 2009
Selecting the target year
Susceptibility of rainbow trout Oncorhynchus mykiss and cherry salmon Oncorhynchus masou against Ichthyophthirius multifiliis
Kim, Yi-Cheong ; Kim, Jin-Do ; Jee, Bo-Young ; Jung, Sung-Hee ; Seo, Jung-Soo ; Park, Sung-Woo ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 193~200
This study was performed to natural infection of Ichthyophthirius multifiliis and mortalities of two fish species were compared at 10
after artificial infection with the parasite. Subsequently compared to motality by the artificial infection with the different parasite orgin of host at 17
using rainbow trout Oncorhynchus mykiss. The prevalence of Ichthyophthirius multifiliis in cherry salmon was higher than that in rainbow trout during the periods of low temperature but no difference in high temperature season. The parasite was not detectable in culturing water and detritus except May that of density has been high. Susceptibility to the parasite was higher in cherry salmon than in rainbow trout at 10
of water temperature but no difference between two fish species at 17
. When rainbow trout were infected with Ichthyophthirius multifiliis from rainbow trout and cherry salmon at 17
, there was no difference in death of host. These results suggest that cherry salmon have higher susceptibility to Ichthyophthirius multifiliis than rainbow trout at low temperature.
Survey of Anisakis spp. infection in wild populations of marine fish caught from coastal areas of Korea
Choi, Hee-Jung ; Jun, Eun-Ji ; Lee, Deok-Chan ; Cho, Mi-Young ; Jee, Bo-Young ; Im, Young-Su ; Park, Myoung-Ae ; Seo, Jung-Soo ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 201~210
Although Anisakis type larvae have been shown to cause various injuries directly or indirectly in humans and animals, the epidemiological studies on these larval infestations are in insufficient state. The status of larval infestation was investigated in 989 fishes of 44 species, which are inhabiting around the east-westernsouth costal area of Korea during the period from March 2007 to February 2008. The Anisakis type larvae were infected approximately 38% (377 fishes) in 989 fishes. Most of the worms were identified as Anisakis simplex type I by morphological finding and 18S ribosomal DNA sequence analysis. In the seasonal variations of infestation, most of the fishes showed higher infestation rate during spring and summer, while the fishes such as herring Clupea pallasii did during winter. From the histopathological studies of infested fishes, it has been observed that Anisakis type larvae are harbouring mainly around the intestinal viscera such as liver, pancreas, stomach, pylolic cecum, and cloaca.
Phylogenetic analysis of marine birnavirus (MABV) isolated from cultured starry flounder Platichthys stellatus and olive flounder Paralichthys olivaceus in Korea
Park, Shin-Hoo ; Park, Myoung-Ae ; Cho, Mi-Young ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 211~218
In this study, we have compared the genome of marine birnavirus (MABV) detected from starry flounder Platichthys stellatus and olive flounder Paralichthys olivaceus. A molecular analysis based on the nucleotide sequence (433 bases) of VP2-NS-VP3 region revealed that MABV (08-KU) from starry flounder showed 98% similarity with MABV Y6 isolated from Yellowtail Seriola quinqueradita in Japan (Accession no: AY283781) and with other aquabirnaviruses identify more than 76%. Comparison with MABV strains (06-KP, 08-KC) from olive flounder and MABV Y6 strain showed 97-98% sequence identities. Phylogenetic analysis was performed in order to examine the relationship among previously determined aquatic birnaviruses isolates showed that MABV and IPNV strains were classified into seven clusters. Three isolates from starry flounder and olive flounder in this study, belong to the genogroup VII including MABV Y6 strain and other aquabirnaviruses isolated from marine fish and molluscan shellfish in Japan. This report is the first description of a MABV from starry flounder in Korea.
Comparison of immunogenecities of three beta-nodavirus proteins, capsid protein, non-structural protein B1 and B2 in olive flounder
Cha, Seung-Ju ; Do, Jeong-Wan ; Ko, Myoung-Seok ; Kim, Jin-Woo ; Park, Jeong-Woo ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 219~228
The genomic and subgenomic RNAs of fish nodavirus encode the four proteins, protein A, capsid protein, non-structural protein B1 and B2. In this study, we describe the immune response of olive flounder Paralichthys olivaceus immunized with live fish nodavirus or recombinant capsid protein, non-structural protein B1 and B2 expressed in E. coli. Nodavirus-infected flounder produced antibodies to capsid protein, B1 and B2 and nodavirus-neutralizing activities were detected in the serum of the nodavirus-infected flounder. The flounder were immunized against the three recombinant proteins of fish nodavirus and the sera from these immunized fishes were assayed for nodavirus-specific antibody by ELISA and a neutralization test. In the immunized flounder, all three recombinant proteins induced the production of similar levels of antibody, but only the antibody to capsid protein significantly neutralized nodavirus. These results indicate that all three nodaviral proteins are immunogenic in flounder, but only the capsid protein can induce neutralizing antibody against nodavirus.
Isolation of marine birnavirus from ascidian Halocynthia roretzi, and its relation with tunic softness syndrome
Song, Jin-Kyung ; Yun, Hyun-Mi ; Choi, Byeong-Dae ; Oh, Myung-Joo ; Jung, Sung-Ju ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 229~237
The causative agent for the tunic softness syndrome of the cultured ascidian Halocynthia roretzi from Jan 1999 to Feb 2009 was identified using virus isolation and polymerase chain reaction (PCR). The pathogenicity of the isolated virus MABV UR-1 strain was determined by experimental infection trials. The cytopathic effects was observed in CHSE-214 cell line at a level 5.1% (4/78) in normal ascidian and 1.8% in abnormal ascidian showing tunic softness syndrome signs. MABV gene was detected in 16.8% (18/107) of normal and 13.1% (5/38) of abnormal organisms by PCR. The ratio of MABV isolation and gene detection was similar level in normal and soft tunic diseased ascidian. Based on the VP2/NS junction region sequences, eight strains of virus isolated from ascidian, were included in the same genogroup with MABV which is originally isolated in wide ranges of marine fish and shellfish species. The UR-1 strain caused 60% mortality (36.5% mortality in control group) by immersion infection and 37% mortality (same mortality in control group) in injection infection indicating no significant differences in infected and control groups. These results suggest that ascidian can act as reservoir of the MABV, and this virus is not directly related with the ascidian mortality.
Effects of the microbiological properties and pathogenicity of Photobacterium damselae subsp. damselae under different culture conditions
Kwon, Mun-Gyeong ; Cho, Byoung-Youl ; Park, Myeong-Ae ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 239~251
The effects of microbiological properties and pathogenicity of Photobacterium damselae subsp. damselae were investigated under different culture conditions, temperature, pH, NaCl and iron concentration on culture media. Favorable conditions for bacterial growth were between 15-30
, pH 5-9, 0-4% NaCl concentration and iron contents of over 10 mM, whereas the bacterial growth was inhibited under iron chelator existence. When P. damselae was cultured in iron-limited tryptic soy broth, total protein concentration of extracellular products, cytotoxic ability of ECPs on cell line, bacterial viability in flounder serum, phospholipase and siderophore activities of ECPs were significantly increased. On the other hand, the activities of P. damselae cultured under iron-added conditions were decreased. In this study, the iron-limited conditions were similar to the host in which iron concentration is low. During infection caused by P. damselae, the conditions could be related to the pathogenesis of the pathogen.
Pathological changes of the heart of olive flounder Paralichthys olivaceus in experimental Streptococcus parauberis infection
Kim, Jin-Woo ; Cho, Mi-Young ; Won, Kyoung-Mi ; Kim, Byoung-Gwan ; Choi, Hee-Jung ; Han, Myoung-Chul ; Park, Myoung-Ae ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 253~262
We conducted bacteriological and histopathological analysis for olive flounder Paralichthys olivaceus after experimental infection with Streptococcus parauberis (FP2284) isolated from diseased olive flounder under different stress conditions. Experimental challenge was performed in healthy flounder (40.4 g in average body weight) by intraperitoneal (i.p.) injection with
CFU/fish under normal (no stress) or netting (for 2 min, twice/day) stress condition. The cumulative mortalities of no-stress and netting stress group were 70% and 95%, respectively. The most prevalent changes observed in experimentally infected flounder were darkness of skin and inflammation of the heart. Severe pericarditis, myocarditis and fibrosis were observed in the heart of the affected flounder. The results of viable counts showed the number of bacteria of the heart tissue was maintained over the
heart for 3 weeks after inoculation. Histological lesions of the heart was more extensive and gradual decrease in bacterial numbers of heart tissue was delayed under stress condition.
The pathogenicity of Streptococcus parauberis isolated from cultured olive flounder Paralichthys olivaceus
Choi, Hee-Jung ; Cho, Mi-Young ; Lee, Jae-Il ; Kwon, Mun-Gyeong ; Choi, Dong-Lim ; Kim, Jin-Woo ; Han, Myoung-Chul ; Lee, Deok-Chan ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 263~273
There is an increasing number of reports describing Streptococcus parauberis as an important pathogen of cultured olive flounder Paralichthys olivaceus and starry flounder Platichthys stellatus in Korea. We tried to determine the effects of water temperature (14
) on the pathogenicity in Streptococcal disease caused by S. parauberis. We have challenged 180 olive flounder by i.p injection to
live cells/fish. Mortality was monitored for 21 days post challenge. And histopathological characterizations as infection degree, tissue degeneration and/or bacterial distribution were investigated with H&E stain and in situ hybridization technique. Fifty percent and 16.7% of mortality occurred within 21 days at 21
water temperature, respectively. In most cases, the typical symptoms of olive flounder infected with S. parauberis were darkness of the skin, lethargy, mild abdominal distension cause by ascites, splenomegaly, congested liver and internal organs paleness. The pericardial sac contained large amounts of cloudy fluid. Numerous whitish nodules, which were variable in size and often confluent, were randomly scattered throughout the myocardium. Especially, pericarditis and/or myocarditis was observed in all tested fishes after death. Positive in reaction with S. parauberis were found in all tissues in situ hybridization analysis. The relative numbers of S. parauberis in heart were much more than in liver, spleen, kidney and stomach. We evaluated that S. parauberis strain causes serious damage in the pericardium, shortness of breath and the blood disorder. Therefore, pericarditis and myocarditis caused by S. parauberis were closely related to mortality of olive flounder.
Family variation in survival of olive flounder Paralichthys olivaceus after Edwardsiella tarda infection in challenge tests
Kim, Hyun-Chul ; Sohn, Sae-Bom ; Jeong, Jong-Geun ; Lee, Jeong-Ho ; Choi, Hye-Sung ; Noh, Jae-Koo ; Park, Choul-Ji ; Min, Byung-Hwa ; Kim, Jong-Hyun ; Kim, Kyung-Kil ; Myeong, Jeong-In ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 275~282
In this study, effect of mating group, mating dam and mating sire was analyzed using olive flounder Paralichthys olivaceus challenged with Edwardsiella tarda. A challenge test was conducted using 1,708 olive flounder fingerlings obtained through
factorial cross from parents selected based on resistant degree. Regarding the means of resistant index, survival rate and days of survival by each mating group, the group(RR) produced by mating among resistant parents were 17.98, 13.77%, and 18.36 days, respectively; the group(SS) produced by mating among susceptible parents were 12.46, 2.71%, and 12.40 days, respectively. This study gives an indication that challenge test results may be successfully used as selection criteria for disease resistance to E. tarda in olive flounder, P. olivaceus.
Effects of extruded pellet and moist pellet feed on health conditions of olive flounder Paralichthys olivaceus
Cho, Mi-Cho ; Oh, Yun-Kyeong ; Park, Shin-Hoo ; Lee, Hae-Young ; Kang, Yong-Jin ; Park, Myoung-Ae ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 283~291
This study was conducted to evaluate the effects of a commercial extruded pellet (EP) and raw fish moist pellet (MP) diet on disease prevalence and serum chemistry of olive flounder Paralichthys olivaceus grown in two commercial-scale aquaculture farms from July to December in 2008. The contents of serum GOT, GPT and glucose in fish fed EP diet (EP group) were higher than those of fish fed the MP diet (MP group). There were no distinct differences in survival rates and mean detection rates of fish pathogens among fish group fed the experimental diets. However, the mean detection rate of fish pathogens in MP group was higher than that of EP group from July to October which are high water temperature season. The dominant pathogens isolated in EP group were Dactylogyrus sp., E. tarda and red sea bream iridovirus (RSIV). On the other hand, Trichodina sp., Streptococcus spp., viral nervous necrosis virus (VNNV) were dominant in MP group.
Effects of low salinity stresses on the physiology of disc abalone, Haliotis discus discus
Jwa, Min-Seok ; Kang, Kyung-pil ; Choi, Mi-Kyung ; Yeo, In-Kyu ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 293~303
Effects of stress on the low salinity stress were examined in the pacific abalone Haliotis discus discus. Changes in survival rate, hemolymph count, antioxidant enzyme activities (catalase: CAT and superoxide dismutase: SOD), respiratory burst activity, phenoloxidase activity, lysozyme activity and expression of heat shock protein 70 (HSP70) mRNA were measured 0, 3, 6, 12, 24 or 48hours after low salinity treatment with 25, 30, 33 and 35 psu. Survival rates of pacific abalone were 100% at 33 and 35 psu, but 93 and 97% at 25 and 30 psu for 48 hours, respectively. Hemolymph counts decreased in the time elapsed-dependent way at all of the experimental groups. At low salinity, 25 and 30 psu, SOD and CAT activity increased compared to the experimental group of 33 psu. Moreover, respiratory burst activities of the pacific abalone seemed to have no effect on low salinity stress at any experimental group. However, phenoloxidase activity is an important component of the defence against pathogen that was decreased in a reduction of salinity dependent way. Lysozyme activity also immediately reduced at 25 psu experimental group for 48 h. The HSP70 mRNA was weakly expressed at 33 psu, but strongly detectable at 25 psu experimental group. The HSP 70 mRNA expression in gill increased in the time elapsed-dependent way at 25 psu experimental group and then recovered at 48 h. These results suggest that low salinity stress give rise to inhibitory action of immune system as a result of the decrease of phenoloxidase and lysozyme activity in the pacific abalone, especially.
Characterization of haemocytes in the surf clam Mactra veneriformis
Yu, Jin-Ha ; Park, Kyung-Il ; Park, Sung-Woo ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 305~316
Haemocyte characterization in the surf clam, Mectra veneriformis was carried out based on morphological, cytochemical, and phagocytic characteristics. The haemocytes were classified into two cell types, granulocytes and agranulocytes on the basis of presence of cytoplasmic granules. Granulocytes were then classified again into 2 types, large eosinophilic granulocytes and small eosinophilic granulocytes after staining with May-Grünwald Giemsa. In electron microscopy, both types of granulocytes contained electron-dense and electron-lucent cytoplasmic granules. Agranulocytes (hyalinocytes) were also divided into two cell types, large agranulocytes and small agranulocytes based on their sizes. Both cell types did not have granules in cytoplasm. Granulocyte and agranulocyte were negative for the enzyme activities of alkaline phosphatase, peroxidase, and
-glucuronidase but positive for phenoloxidase and acid phosphatase activities. Both types of haemocytes have phagocytic activity, with the exception of small agranulocyte, and granulocytes seemed more active in this respect than agranulocytes. This present study is the first study to characterize haemocytes of M. veneriformis.
Antimicrobial and antioxidant activity of some Indian medicinal plants for the protection against fish pathogenic bacteria
Harikrishnan, Ramasamy ; Jawahar, Sundaram ; Kim, Man-Chul ; Kim, Ju-Sang ; Jang, Ik-Soo ; Balasundaram, Chellam ; Heo, Moon-Soo ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 317~326
This study has shown the screening of anti-bacterial activity of three Indian medicinal plant choloroform : methanol (50:50) solvent leaf extracts (i.e. Azadirachta indica, Ocimum sanctum, and Curcuma longa) with different concentrations (10, 5, 2.5, 1.25, 0.625, 0.312, and 0.156 mg/ml) under in vitro conditions against fish pathogenic bacteria, Aeromonas hydrophila, Streptococcus iniae, Vibrio harveyi, V. anguillarum, and Edwardsiella tarda isolated from olive flounder farms, Jeju Island, South Korea. The anti-microbial activity of the A. indica and O. sanctum extracts yielded the zones of growth inhibition (ZI) was 3 and 1mm against A. hydrophila at concentration of 0.156 mg/ml when compared to that of tetracycline standard (3 mm). At highest concentration (10 mg/ml) of A. indica, O. sanctum, and C. longa, high inhibition was 9, 7, and 6 mm when compared to that of tetracycline (11 mm) against A. hydrophila. The minimum inhibitory concentration (MIC) of A. indica, O. sanctum, and C. longa at 0.156 mg/ml that yield 9, 10, and 13 CFU/ml for A. hydrophila, 16, 22, and 25 CFU/ml for S. iniae and 18, 22, and 23 CFU/ml for E. tarda compared to the tetracycline. At highest concentration (10 mg/ml) of the three extracts was better inhibiting the growth of A. hydrophila, S. iniae and E. tarda. A. indica, O. sanctum, and C. longa were determined to the potential antioxidant activityon the basis of their scavenging activity of the stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical. A. indica extract was 0.625 mg/ml which indicated that the strong anti-oxidant activity. However, O. sanctum and C. longa extracts showed weak anti-oxidant activity at this concentration. Hence, in vitro assay among the pathogens, A. hydropila is better inhibitory activity of the extracts. It is evident that the Indian medicinal plants extracts were subjected to its effectiveness against A. hydrophila, S. iniae, and E.tarda at low concentrations. The obtained results in the present study suggested that the Indian plant extracts is a prevention tools for Korean olive flounder aquaculture pathogens and its need further advance investigation.
The efficacy and influence on growth of olive flounder Paralichthys olivaceus vaccinated against Edwardsiella tarda and Streptococcus iniae
Kim, Myoung-Sug ; Kim, Kyoung-Duck ; Kim, Kang-Woong ; Park, Myoung-Ae ; Kim, Jin-Woo ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 327~334
This study was performed to verify the efficacy and safety of vaccine mixed formalin killed Edwardsiella tarda and formalin killed Streptococcus iniae in olive flounder Paralichthys olivaceus. Sera were obtained from each group at 2, 4, 6 and 8 weeks after vaccination and agglutination titers to E. tarda and S. iniae were determined using the microtiter method. Three weeks after vaccination, the fish challenged by intraperitoneal injection of E. tarda or S. iniae and immunization resulted in a relative percentage survival (RPS) of above 81.0-92.8%. Neither the groups vaccinated nor control groups resulted in significantly reduced weight gain 140 days post-vaccination, but differences between the groups vaccinated and control groups were found at the early days post-vaccination. This results suggest that it is possible to prevent edwardsiellosis and streptococosis after a vaccination without affecting growth of olive flounder.
Evaluation of the stability of IgM and specific antibody response of sevenband grouper Epinephelus septemfasciatus for application of antibody-detection ELISA
Kim, Chun-Seob ; Jang, Min-Seok ; Kim, Wi-Sik ; Kim, Jong-Oh ; Kim, Du-Woon ; Kim, Do-Hyung ; Han, Hyun-Ja ; Jeong, Sung-Ju ; Oh, Myung-Joo ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 335~342
The stability of immunoglobulin M (IgM) on different serum storage conditions and specific antibody response were tested using the serum collected from sevenband grouper Epinephelus septemfasciatus by enzyme-linked immunosorbent assay (ELISA). To test the effect of storage temperature and duration, sevenband grouper antiserum against bovine serum albumin (BSA) was stored at -80, -20 or 4
for 1, 34, 61 or 119 days. In addition, to test the effect of repeated freeze-thawing condition, the anti-BSA fish serum was frozen at -20 and -80
and then thawn and frozen for 1, 5 or 10 times repeatedly. Consequently, no significant difference was found in ELISA optical density (O.D.) values of sera for the above mentioned storage conditions: different temperatures (-80, -20 and 4
), durations of storage (1, 34, 61 and 119 days), and repeated thaw-freeze cycles (1, 5, and 10 times), indicating that IgMs of test fish were stable. The specific antibody response of sevenband grouper was observed after BSA-immunization of the test fish reared at 20
. At the rearing temperature of 20
, the specific antibody against BSA first appeared at 14 days and maximum antibody titer was observed between 21 and 28 days, while at the rearing temperature of 25
, specific antibody appeared at 7 days and maximum antibody titer was observed between 14 and 21 days. In conclusion, the rearing temperature at 25
gave a faster and higher specific antibody response than at 20
and the specific antibody response maintained for approximately 2 months at 20℃ and 25
Molecular identification and expression analysis of a natural killer enhancing factor-A from black rockfish Sebastes schlegelii
Lee, Jeong-Ho ; Kim, Joo-Won ; Park, Chan-Il ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 343~352
Natural-killer-cell-enhancing factor (NKEF) belongs to the newly defined peroxiredoxin (Prx) family. It was originally isolated from human erythroid cells. The black rockfish NKEF cDNA was identified through the expressed sequence tag (EST) analysis of PBLs libraries. The full-length NKEF cDNA was 1433 bp long and contained an open reading frame (ORF) of 594 bp that encoded 198 amino-acid residues. The 5' UTR had a length of 39 bp, and the 3’UTR 800 bp. The deduced amino-acid sequence of the black rockfish had a density 93.4, 92.9, 87.8, 85.8, 84.8, 83.8, 80.3, 79.7, 77.2, and 75.2% that of the pufferfish, olive flounder, channel catfish, zebrafish, chicken, common carp, Myotis lucifugus, cattle, human PrxI, rat PrxI, human NKEF-A, and Xenopus tropicalis, respectively. The NKEF gene was expressed in all the tissues of the black rockfish. The RT-PCR indicated that the NKEF transcripts were predominantly in the spleen and gill, less dominantly in the PBLs, head kidney, trunk kidney, and liver, and least in the intestine and muscles. This is the first report on the existence of the NKEF-A gene in black rockfish.
Expressed sequence tags analysis of immune-relevant genes in rock bream Oplegnathus fasciatus peripheral leukocytes stimulated with LPS
Lee, Jeong-Ho ; Noh, Jae-Koo ; Kim, Hyun-Chul ; Park, Choul-Ji ; Min, Byung-Hwa ; Choi, Sang-Jun ; Myeong, Jeong-In ; Park, Hyung-Jun ; Park, Chan-Il ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 353~366
We constructed a rock bream Oplegnathus fasciatus leukocyte cDNA library and a total of 795 expressed sequence tag (EST) clones were generated. Gene annotation procedures and homology searches of the sequenced ESTs were locally done by BLASTX for amino acid similarity comparisons. Of the 795 EST clones, 491 different ESTs showed significant homology to previously described genes while 304 ESTs were unidentified, hypothetical, or unnamed proteins. Encoding 121 different sequences were identified as putative bio-defense genes or genes associated with immune response.
The effect of omae Prunus mume extract on the immune response and growth rate of Japanese eel Anguilla japonica
Kim, Jin-Do ; Kim, Yi-Cheong ; Kim, Gwang-Seok ; Woo, Sung-Ho ; Park, Sung-Woo ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 367~374
The effects of extacts of Omae Prunus mume were tested on non-specific immune response of Japanese eel Anguilla japonica. Lysozyme activity in the serum of the administrated group was presented higher than control, whereas no difference was lysozyme in the mucus or bacterial reaction of the serum. The lysozyme activity of the serum was increased 2 weeks after administrated and highest of that was 5 weeks later. Results of the experimental challenge of Edwardsiella tarda and Aeromonas hydrophila were presented administrated group was higher than control. The results of both groups in the growth rate were same.
Integration of a target gene into chromosomal genome of BF-2 cells using UV-inactivated snakehead retrovirus (SnRV)
Kwon, Se-Ryun ; Nishizawa, Toyohiko ; Yoshimizu, Mamoru ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 375~382
Integration and expression of a target gene into chromosomal genomes of host cell by retrovirus mediated gene transfer system usually require complicate and laborious procedures. In the present study, we investigate a simple method to integrate a target gene into genome of BF-2 cells using ultraviolet (UV)-inactivated snakehead retrovirus (SnRV), a fish retrovirus. First of all, an optimization of transfection condition was determined with BF-2 cells using Lipofectamine 2000 and Transome. Using 0.5
Lipofectamine 2000 resulted in 33.8, 40.6 and 40.2% of transfection efficacy with high survival rate (minimum 80%) in 0.5, 1 and 2
DNA, respectively, and those of Transome were all less than 5%. It was confirmed that UV-treatment for 5 min was enough to inactivate infectivity of SnRV. Next, a cassette composed of GFP (green fluorescent protein) gene flanked by LTR (long terminal repeats) sequences derived from SnRV was constructed and transfected into BF-2 cells followed by treatment with UV-inactivated SnRV for optimization of integration and expression of the cassette gene. As the results, the fluorescence was expressed in BF-2 cells treated with UV-inactivated SnRV 3 and 5 times, while there was no expression in BF-2 cells with once and non treatment. Accordingly, it was confirmed that GFP gene was integrated into chromosomal genome of BF-2 cells with UV-inactivated SnRV.
An EST-based approach for identifying genes expressed in the gills of olive flounder Paralichthys olivaceus
Lee, Jeong-Ho ; Noh, Jae-Koo ; Kim, Hyun-Chul ; Park, Choul-Ji ; Min, Byung-Hwa ; Kim, Young-Ok ; Kim, Jong-Hyun ; Kim, Kyung-Kil ; Kim, Woo-Jin ; Myeong, Jeong-In ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 383~389
Analysis of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and development of resources useful for functional genomics studies. As part of studies on the immune system of olive flounder, a total of 251 EST sequences from gill cDNA library were generated to identify and characterize important genes in the immune machanisms of olive flounder. Of the 251 clones, 126 clones (50.2%) were identified as orthologues of known genes from olive flounder and other organisms. Among the 126 EST clones, 16 clones (12.7%) were representing 9 unique genes identified as homologous to the previously reported olive flounder ESTs, 100 clones (79.4%) representing 103unique genes were identified as orthologs of known genes from other organisms. We also identified several kinds of immune associated proteins, indicating EST as a powerful method for identifying immune related 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.
Discrepancies in genetic identification of fish-derived Aeromonas strains
Han, Hyun-Ja ; Kim, Do-Hyung ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 391~400
Genetic identification of 17 fish-derived Aeromonas strains was attempted using 5 housekeeping genes. 16S rRNA, gyrB, rpoD, dnaJ and recA genes from the 17 strains were amplified, and total of 85 amplicons were sequenced. DNA sequences of the strains and type strains of the 17 Aeromonas homology groups were used for genetic identification and phylogenetic analyses. None of the strains was identified as a single species using the 16S rRNA gene, showing the same identities (average = 99.7%) with several Aeromonas species. According to gyrB, rpoD, dnaJ, and recA, 9 strains and RFAS-1 used in this study were identified as A. hydrophila and A. salmonicida, respectively. However, the other strains were closely related to 2 or more Aeromonas species (i.e., A. salmonicida, A. veronii, A. jandaei, A. media and A. troda) depending on the genetic marker used. In this study, gyrB, rpoD, dnaJ and recA gene sequences proved to be advantageous over 16S rRNA for the identification of field Aeromonas isolates obtained from fish. However, there are discrepancies between analyses of different phylogenetic markers, indicating there are still difficulties in genetic identification of the genus Aeromonas using the housekeeping genes used in this study. Advantages and disadvantages of each housekeeping gene should be taken into account when the gene is used for identification of Aeromonas species.
Isolation and identification of Flavobcterium succinicans from anadromous ayu Plecoglossus altivelis
Lee, Chang-Hoon ; Kim, Pil-Youn ; Lim, Bong-Soo ; Oh, Duck-Chul ; Kang, Bong-Jo ;
Journal of fish pathology, volume 22, issue 3, 2009, Pages 401~406
On May in 2008, mortality of anadromous Ayu Plecoglossus altivelis was observed on the Gangjeong river in Jeju. Major symptoms of the infected fish were mouth rot and skin ulcer. The causative agent was suspected as gliding bacteria. After culture on Shu-Shott and R2A media, we isolated bacterium belonging to the Flavobacterium from ayu with symptoms. As a result, the bacterium was identified as Flavobacterium succinicans JMFL55 by 16S rDNA sequence alignment with F. succinicans DSM 4002(98.27% similarity, GenBank accession NO. AM230492).