ヘテロボツリウム孵化幼生を用いたトラフグへの標準化感染実験 Standardized Method for Experimental Infection of Tiger Puffer Takifugu rubripes with Oncomiracidia of Heterobothrium okamotoi (Monogenea : Diclidophoridae) with Some Data on the Oncomiracidial Biology

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Author(s)

    • 千ヶ崎 学 CHIGASAKI Manabu
    • Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo
    • 中根 基行 NAKANE Motoyuki
    • Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo
    • 小川 和夫 [他] OGAWA Kazuo
    • Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo
    • 若林 久嗣 WAKABAYASHI Hisatsugu
    • Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo

Abstract

蛍光色素 CFSE で生体染色したヘテロボツリウム孵化幼生を0歳トラフグに感染させた。 CFSE は孵化幼生の感染能力に影響しなかった。 0―4日齢の孵化幼生を異なる濃度(1L, 1尾当たり 29,58,116虫)と接触時間(1―10時間)で感染させ, 以下の結果を得た。 孵化幼生の感染能力は経日的に低下し, 2日齢以降はほとんど感染しなかった。 濃度は着定率に影響しなかった。 着定率は3時間以降は有意に増加しなかった。 従って, 孵化後1日以内の幼生に3時間接触させるのを標準感染法とした。

0-year-old tiger puffer, Takifugu rubripes, were exposed to suspensions of Heterobothrium okamotoi oncomiracidia under different conditions. A fluorescent dye, CFSE (5 (6) -carboxy fluorescein diacetate, succinimidyl-ester), was used as a mean of labeling oncomiracidia to find them with a fluorescence Microscope immediately after settlement on the host. The labeling procedure did not affect the infectivity of the oncomiracidium. When fish were exposed to labeled oncomiracidia for 1, 3, 5 or 10 h, the settlement rate on the gills increased with increasing exposure time up to 3 h. When similarly exposed to labeled oncomiracidia of different ages (0-4 days after hatching), infectivity of the oncomiracidium decreased rapidly with increasing parasite age, and most oncomiracidia of 2 days or older failed to infect the gills. When exposed to 29, 58 and 116 labeled Oncomiracidia/ L/ fish and observed under fluorescence stereomicroscopy immediately after exposure, the settlement rates on the body surface and on the gills were not different among the 3 concentrations tested. The settlement rate on the body surface was always higher than that on the gills, irrespective of the difference in concentration. Thus, infection methods using tiger puffer and H. okamotoi oncomiracidia were standardized in terms of exposure time (3 h), parasite age (within 24 h after hatching), and parasite concentration (up to 116 oncomiracidia/ fish). CFSE-staining is suitable for the study of early stages of parasite settlement on the host fish.

Journal

  • Fish Pathology

    Fish Pathology 35(4), 215-221, 2000-12-15

    The Japanese Society of Fish Pathology

References:  14

Cited by:  6

Codes

  • NII Article ID (NAID)
    10019222497
  • NII NACSIS-CAT ID (NCID)
    AN00063165
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    0388788X
  • NDL Article ID
    5605421
  • NDL Source Classification
    ZR4(科学技術--生物学--動物)
  • NDL Call No.
    Z18-994
  • Data Source
    CJP  CJPref  NDL  J-STAGE 
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