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Isolation of fish macrophage and its physiology

Research Project

Project/Area Number 09660207
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General fisheries
Research InstitutionMiyazaki University

Principal Investigator

ENDO Makoto  Miyazaki Univ., Fac. Agriculture, Assoc. Professor, 農学部, 助教授 (80128355)

Co-Investigator(Kenkyū-buntansha) YOSHIDA Terutoyo  Miyazaki Univ., Fac. Agriculture, Assoc. Professor, 農学部, 助教授 (20240294)
IIDA Takaji  Miyazaki Univ., Fac. Agriculture, Professor, 農学部, 教授 (70159557)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1999: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1998: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
KeywordsFish / Macrophage / swim bladder / inflammation / phagocytosis / cytokine / ストレス / 分離
Research Abstract

Neutrophils of Tilapia Oreochromis niloticus were collected from the swim bladder 24 hrs after infection with 1 mg/fish of formalin-killed Eschericia coli in Hanks' balanced solution. The neutrophils were sonicated using an ultrasonic processor for 60 sec. To collect inflammatory macrophages, the fish were injected with 0.2 mL of the neutrophil cytolysates into the swim bladders, and cells that exuded into the organ were harvested at 4 days after injection. The cells were smeared and stained with Giemsa, non-specific esterase, and peroxidase stains to identify the cell types. Macrophages occupied 40% of the exudate cells. To purify the inflammatory macrophages, the exudate cells were incubated in L-15 medium 25℃ for 48 hrs or subjected to a gradient-centrifuge procedure with Ficoll (1.065 in gradient, at 1,500 rpm for 30 min). Both the methods enabled to purify the macrophages more than 90%.
In order to examine the macrophage physiology, the phagocytic activities and various cytokines levels (TNF-α, IL-1β, IL-10, and IL-12) with a ELISA method using rabbit IgG against them were surveyed on the macrophages. Besides, immunostimulants like glucan raised their physiological parameters of macrophages. Tilapia were kept under self-feeding for 1 week to make lower stressful condition. An analysis on the blood cortisol confirmed the lower cortisol level in the fish kept under the self-feeding than the ones kept under the usual feeding, meaning the self-feeding provides the low stress condition to the fish. The macrophages collected from the fish kept under the self-feeding showed the higher phagocytotic activities and cytokine productivities of macrophages. Thus, it appears that the stressful condition influences the macrophage physiology.

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] Chen, S. -C., T. yoshida, S. Adams, and R. richards: "Non-specific immune response of Niltilapia, Oreochromis nilotica, to the extracellular product of Myobacterium, spp. and to various adjuvants"J. Fish Diseases. 21. 39-46 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuyama, T., Tilida, and M. Endo: "Isolation of inflammatory niacrophages from swim bladder of tilepia"Fish Pathology. 34. 83-84 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 松山知正、飯田貴次、延東真: "鰾を用いた白血球の集収法"海洋. 号外14. 140-144 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Chen,S.-C., T.Yoshida, S.Adams, R.Richards: "Non-specific immune response of Nile tilapia, Oreochromis nilotica, to the extracellular products of Mycobaterium spp. and to various adjuvants"Journal of Fish Diseases. 21(1). 39-46 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuyama,T., T.Iida, and M.Endo: "Isolation of inflammatory macrophages from swim bladder of tilapia"Ocean (kaiyo). 14(supplement). 140-144 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuyama,T., T.Iida, and M.Endo: "Isolation of leukocytes using swim bladder"Fish Pathology. 34(2). 83-84 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T.Matsuyama,T.Iida,and M.Endo: "Isotation of Inflammatory Macrophages from Swim bladder of Tilapia"Fish Pathology. 34・2. 83-84 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Chen,S-C,T.Yoshida,S,Adams,R.Richards: "Non-specific immuneresponse of Nile tilapia,Dreothromrs nilotica,to the extracellular products of Mycobacterium spp,and to various adjuvants" Journal of Fish Disease. 21・1. 39-46 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 松山・飯田・延東: "鰾を用いた白血球の収集法" 海洋. 号外・14. 140-144 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Chen,S-C,et al.: "Non specific immune response of Nile tilapia,Oneochromis milotica,to the extracellular products of Mycobac terium spp.and to various adjuvants" Journal of Fish Disease. 21・1. 39-46 (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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