• Title/Summary/Keyword: mud fish

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Production of Transgenic Homozygous Diploid in Mud Loach(Misgurnus mizolepis) I. Transfer of Luciferase Gene and Evaluation of Mud Loack Expression Vector

  • Nam Yoon Kwon;Kim Moo-Sang;Lee Hyung-Ho;Kim Dong Soo
    • Journal of Aquaculture
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    • v.9 no.3
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    • pp.293-300
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    • 1996
  • Validities of several gene transfer methods including microinjection, electroporation and lipo-fection with luciferase gene (pRSVL), and effectiveness of mud loach expression vector which contains ARS from mud loach on production of transgenic mud loach were evaluated. Microiniection revealed the $0\~8\%$ of transgene incidence in 2-week-old fish with significant mosaicism. Electroporation and lipofection of mud loach sperm also successfully introduced the transgene into sperm cells, and transferred the foreign DNA into zygote. Gene transfer by electroporation and lipofection showed a range of $0\~28\%$ and $0\~48.1\%$ of transgene incidence, respectively in newly hatched larvae, altough most DNA introduced were gradually degraded with the development of fish. Microinjections of mud loach expression vector caused a significantly reduced survival rate of mud loach embryos with severe teratogenic effects, and ARS/Luc transgene could not be detected in normally developed fish after microinjection.

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Genetic breeding of fast-growing autotransgenic mud loach, Misgurnus mizolepis

  • Kim, Dong-Soo;Nam, Yoon-Kwon
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.10a
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    • pp.25-25
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    • 2001
  • Generation of transgenic fish acquiring the ability to express desirable phenotypes offers new possibilities for addressing fundamental biological questions, and can also attribute to enhanced aquaculture productivity. I describe here the recent research progress in my laboratory with particular emphasis on the development of fast-growing autotransgenic fish and its chromosome-set manipulation using our experimental organism, the mud loach, Misgurnus mizolepis. (omitted)

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Salt treatment for recovery of the mud loach, Misgurnus mizolepis from transport stress

  • Yu, Jin-Ha;Kim, Dae-Hyun;Han, Jung-Jo;Park, Sung-Woo
    • Korean Journal of Veterinary Research
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    • v.56 no.4
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    • pp.215-221
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    • 2016
  • Due to the shortage of the fingerling/juvenile mud loach, Misgurnus mizolepis in Korea, these fish have been imported from China. However, the mortality rate during and after their transportation is very high. In this study, we examined various physiological and histological parameters to evaluate the effect of salt treatment on the survival and recovery of mud loaches in holding farms during the quarantine process. Glucose, osmolality, $Na^+$, $Cl^-$, and histological changes were assessed for three different salinities. Non-treated fish (control 0.0%) exhibited lower levels of osmolality, and $Na^+$ and $Cl^-$ concentrations compared with those kept in solar salt solution (0.5% and 1.0%). Glucose levels in control fish were higher than those in fish exposed to 0.5% and 1.0% solar salt solution. Histologically, control fish showed thinner epidermis of skin, branchial hyperplasia and lamellar fusion with an abundance of eosinophilic granule cell-like cells. After solar salt solution treatment, damaged gill structures in the fish almost recovered within 5 days. The present study demonstrates that mud loaches transported from China suffer from skin and gill damage and physiological dysfunction which may increase the mortality and morbidity. Moreover, saline treatment might alleviate the stress responses and ionic/osmotic imbalances, and help heal gill damage.

Growth Performance of Transgenic Mud Loach Misgurnus mizolepis Carrying a GH Transgene Driven by Mud Loach C-Type Lectin Regulator

  • Song, Ha-Yeon;Kim, Dong-Soo
    • Fisheries and Aquatic Sciences
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    • v.15 no.1
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    • pp.43-47
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    • 2012
  • Growth hormone (GH) transgenesis in fish has the potential to improve aquaculture efficiency and capacity. However, many fast-growing transgenic fish have experienced side effects caused by excess GH expression. To overcome this unwanted issue associated with several GH transgenic mud loach Misgurnus mizolepis lines carrying GH construct driven by a strong ${\beta}$-actin regulator ($pml{\beta}$-actGH), we performed an alternative version of GH autotransgenesis using a weaker but more stable regulator, the mud loach lectin promoter. GH transgenesis with a pmlectGH construct consisting of the mud loach GH gene driven by the 2.3-kb lectin promoter exhibited significant growth stimulation. However, the extent of the growth acceleration in pmlectGH transgenics (six times maximum when assessed 2 months post hatching) was much less than that in transgenic individuals carrying the $pml{\beta}$-actGH construct. Additionally, the extraordinary gigantism that was common in $pml{\beta}$-actGH-transgenic mud loaches was diminished in transgenic loaches harboring the pmlectGH construct. Transgenic founders (pmlectGH) successfully transmitted their transgene into the next generation with up to 41% frequency. Growth stimulation also persisted in the transgenic F1 strains, with a seven-fold increase in maximum body weight at 6 months of age.

Screening of Potential Stress-Responsive and Immune-Related Genes by Expressed Sequence Tags in Mud Loach (Misgurnus mizolepis)

  • Nam, Yoon-Kwon;Kim, Dong-Soo
    • Journal of fish pathology
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    • v.15 no.2
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    • pp.83-92
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    • 2002
  • EST analysis was performed to identify stress-responsive and immune-related genes from mud loach (Misgurnus mizolepis), cDNA libraries were constructed with liver, intestine and kidney tissues and randomly chosen clones (216 for liver, 198 for intestine and 224 for kidney) were subjected to automated sequence analysis. Of 638 clones sequenced in totlal, approximalely 25% of ESTs was novel sequences (no match to GenBank) or sequences with high homology to hypothrtical/unknown genes. Several potential stress-responsive biomarker and/or immure-related genes were identified in all the tissues examined. It included lectin, MHC class I/II proteins, proteinase inhibitors, superoxide dismulase, catalase, glutathionc-S. transferase, heat-shock protein, warm temperature acclimation protein, complements, methylrransferasc, zinc finger proteins, macrophage maturation associated protein, and others. This information will offer new possibilities as fundamental baseline data for the molecular genetics and breeding of this species with an emphasis on the development of stress. (and disease)-resistsnt fish.

An Overturn disease of Cultured Mud Loach (Misgurnus mizolepis) (미꾸라지 (Misgurnus mizolepis)의 뒤집힘병에 관한 연구)

  • Park, Seong-U;Park, Hyeon-Tae;Choe, Seon-Nam
    • Journal of fish pathology
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    • v.17 no.1
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    • pp.21-28
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    • 2004
  • In late summer of 2001, a new disease "overturn disease" occurred among mud loach (Misgurnus mizolepis) cultured in a farm Chonbuk Province. The fish exhibited a lethargic, erratic and spiral swimming at the water surface. Affected fish showed swollen abdomen with accumulation of air bubbles in the intestinal canal. The bubbles were much larger in the anterior part than in the posterior part. Hematological and histopathological examinations were done. No significant differences were found in RBC count, Ht value and Hb concentration between the diseased and the healthy fish. Gill filaments showed proximal hyperplasia of the respiratory epithelia and fusing of the gill lamella. Hemorrhage and necrosis occurred in the intestinal epithelia and within the lamina propria, and the mucosal epithelia were separated from the muscularis. Hepatocytes underwent atrophy. In the fish experimentally tied between the annal fin and the anus by a thread to confirm accumulation of air bubbles within intestine, accumulation of air bubbles was confirmed. This disease in mud loach appeared to be induced by circulatory disturbances in the intestinal wall owing to accumulation of air bubbles in the alimentary canal after fed high protein diet throughout all culturing period.

Isolation of Aeromonas sobria from Cultured Mud Loach, Misgurnus mizolepis (양식 미꾸라지(Misgurnus mizolepis)로 부터 Aeromonas sobria 검출)

  • Yu, Jin-Ha;Park, Sung-Woo
    • Journal of fish pathology
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    • v.21 no.1
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    • pp.21-27
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    • 2008
  • Mass mortality occurred in mud loaches, Misgurnus mizolepis, cultured in ponds located in Kunsan. External signs of affected fish showed hemorrhage of skin and fins, Internally, pale liver with congestion, enlarged kidney, and spleen and enteritis exhibited. Causative bacteria isolated from liver, spleen, and kidney of the disease fish. In biochemical tests, the isolates were similar with those of the reference strains, A. sobria. The aerolysine gene from the present isolate was amplified PCR with the primer SOBF and SOBB for A. sobria. The isolate was identified as A. sobria on the basis of those tests. In virulence test, the present isolate resulted in the development of clinical signs identical to those in naturally infected fish. The present results conclude that the present isolate is A. sobria and can be a pathogen which causes motile aeromonad septicemia to mud loach.

Triploid hybridization as a reproductive containment method of genetically modified fish, exemplified by fast-growing transgenic mud loach

  • Nam, Yoon-Kwon;Park, In-Seok;Kim, Dong-Soo
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.41-41
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    • 2003
  • Transgenic triploid hybrid between fast-growingtransgenic mud loach (Misgurnus mizolepis) males and cyprinid loach (M. anguillicaudatus) females were generated and their performance on growth, feed conversion ability and reproduction were evaluated. Although the growth accelerations of diploid and triploid transgenic hybrids were not as much as those of original transgenic mud loaches, they still represented persistent growth stimulation ranging 11 to 28 fold when compared to their non-transgenic counterparts, with significantly improved feed conversion efficiency up to 2-fold (compared to non-transgenic hybrid) and 1.5-fold (compared to non-transgenic mud loach) in maximum. The gonad development of diploid hybrids was fertile in histological views regardless of transgenic genotypes but the extent of developmentin hybrid fish were less than mud loach diploids at the same age. On the other hands, very stringent sterility was obtained in both sexes of the triploid hybrid transgenics: ovary and testis from transgenic triploid hybrids were significantly depressed and any notable sign for maturation to ovum or spermatids was not detected. No viable embryo was obtained in a fertilization trial using the suspension prepared from the minced testes of transgenic triploid hybrids. This study may indicate the potential usefulness of triploid hybridization as a mean for reproductive containment of transgenic mud loach.

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The Light Quality Dependence of Photoresponse of Mud Fish (Misgurunus mizolepis $G{\"{u}}nther$)and the Chromophores Photogenerating Active Oxygen in its Skin Tissues (미꾸라지 광반응의 광질 의존성과 피부조직의 내생 광증감제)

  • Boo, Yong-Chool;Jung, Jin
    • Korean Journal of Environmental Agriculture
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    • v.11 no.3
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    • pp.253-260
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    • 1992
  • A photoresponse that results in organisms dispersing from a region of bright light. generally termed photodispersal, is frequently observed in some fishes notably including mud fish (Misgurunus mizolepis G${\"{U}}$NTHER). The primary assumption for this study was that the photodispersion may result from the behavioral strategies of fishes aimed to avoid illumination conditions that could injure the cells in skin tissues via photodynamic sensitization reactions. Here we present some preliminary results that seem to support this assumption : (1) the locomotive action of dark-adapted mud fish was triggered by the onset of illumination with light : (2) blue light (400-500nm) was much more effective in bringing about the locomotive activity than yellow (550-650nm) and red (650-800nm) lights : (3) two blue light absorbing pigments, which photogenerate activated oxygen species, were separated from the skin tissues of mud fish, one of these being identified as riboflavin.

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Altered expression of mud loach (Misgurnus mizolepis; Cypriniformes) hepcidin mRNA during experimental challenge with non-pathogenic or pathogenic bacterial species

  • Lee, Sang-Yoon;Kim, Dong-Soo;Nam, Yoon-Kwon
    • Journal of fish pathology
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    • v.24 no.3
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    • pp.279-287
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    • 2011
  • Transcriptional response patterns of mud loach (Misgurnus mizolepis; Cypriniformes) hepcidin, a potential ortholog to human hamp1, in response to experimental challenges with non-pathogenic and pathogenic bacterial species were analyzed based on the semi-quantitative reverse transcription-PCR assay. Mud loach hepcidin transcripts were much more preferentially induced by pathogenic bacterial species (Edwardsiella tarda and Vibrio anguillarum) causing apparent pathological symptoms than by non-pathogenic species (Escherichia coli and Bacillus thuringiensis) displaying neither clinical signs nor mortality. However in overall, the induced amounts of hepcidin transcripts were positively related with the number of bacterial cells delivered in both pathogenic and non-pathogenic bacterial species. Inducibility of hepcidin transcripts were variable among three tissues examined (liver, kidney and spleen) in which kidney and spleen were more responsive to the bacterial challenge than liver. Time course expression patterns of hepcidin mRNAs after challenge were different between groups challenged with pathogenic and non-pathogenic species, although the overall pattern of hepcidin expression was in accordance with that generally observed in battery genes appeared during early phase of inflammation. Fish challenged with E. coli (non-pathogenic) showed the significant induction of hepcidin transcripts within 24 hr post injection (hpi) but the level was rapidly declined to the basal level either at 48 or 96 hpi. On the other hand, hepcidin transcript levels in E. tarda (pathogenic)-challenged fish were continuously elevated until 48 hpi, then downregulated at 96 hpi, although the level at 96 hpi was still significantly higher than control level observed in non-challenged fish. This expression pattern was consistent in all the three tissues examined. Taken together, our data indicate that hepcidin is tightly in relation with pathological and/or inflammation status during bacterial challenge, consequently providing useful basis to extend knowledge on the host defensive roles of hepcidin under infectious conditions in bony fish.