스쿠티카충 Miamiensis avidus에 대한 넙치의 면역반응
Immune Response of Olive flounder, Paralichthys olivaceus against Miamiensis avidus (Ciliophora: Scuticociliatida)
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Miamiensis avidus는 넙치에 폐사를 일으키는 스쿠티카 섬모충이다. M. avidus에 대한 넙치의 면역반응을 측정하기 위하여 FKC (formalin killed cell)를 마리당 2.6 × 106 cells을 복강주사하고, FKC를 초음파처리한 것을 2.4 × 106 cells/㎖로 14.9 cm (26.8 g)의 넙치에 2주 간격으로 두 번 면역하였다. 면역한 넙치의 항체는 충체의 운동성둔화 (immobilization)와 응집 (agglutination)을 유발하였다. 면역한 그룹은 면역하지 않은 그룹보다 높은 ELISA항체가를 나타내었다. 또한, 복강주사로 면역한 넙치에서 유의적으로 높은 phagocytic ratio, phagocytic index와 주화활성을 나타내었다. FKC의 면역방어효과를 검증하기 위하여 8.6 cm (6.3 g)의 넙치에 마리당 1.43 × 105 cells을 복강주사면역하고 2.2 ×105 cells/㎖의 농도로 침지면역했다. 2주 간격으로 두 번 면역하고 2.0 × 104과 2.0 × 103 cells/㎖ 농도로 침지 공격 감염시킨 후 폐사율을 관찰하였다. 감염시킨 3주 후 폐사율은 복강주사구에서 감소하였고 침지면역구는 폐사가 지연되었다. 위의 결과로부터 M. avidus의 불활화백신을 넙치에 면역하면 충에 대한 특이면역이 증강되며 증강된 면역반응이 방어력을 제공했을 것으로 생각되었다.
Abstract
Miamiensis avidus is a scuticociliate causing mortality in olive flounder Paralichthys olivaceus. To evaluate immune response of olive flounder against M. avidus, 2.6×106cells/fish of Formalin killed cell (FKC) was intraperitoneally (i.p.) injected, and 2.4 × 106cells/㎖ of sonicated FKC was immersion immunized to 14.9 cm (26.8g) fish. Fish were immunized 2 times with 2 weeks intervals. Antiserum from immunized fish caused agglutination and immobilization of the ciliate. In ELISA test, immunized group exhibited higher titers than control group. In addition, i.p. immunized fish showed higher phagocytic ratio, phahgocytic index and chemotaxic activity. To evaluate in vivo efficacy of FKC on fish, 1.43 × 105cells/fish was i.p.. injected, and 2.2 × 105 cells/㎖ of sonicated cell was immersed into 8.6cm (6.3g) fish. After 2 times immunization with 2 weeks intervals, fish were infected with 2.0 × 104 and 2.0 × 103 cells/ml of live ciliates by immersion. After 3 weeks, cumulative mortality was lower in the i.p. immunized group and mortality was delayed in immersion immunized group. In conclusion, specific immune response of oliver flounder against M. avidus was elevated after immunization and these immune response may prevent and/or delay for the M. avidus infection to olive flounder.
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