Vibrio scophthalmi에 대한 넙치,Paralichthys olivaceus의 면역반응
- Abstract
- Vibrio scophthalmi는 Vibrionaceae에 속하는 Gram-negative bacteria이며,우리 나라 넙치 양식장에서 새로운 피해 원인으로 대두되고 있다. 본 연구에서는 V. scophthalmi 강독성 균주와 약독성 균주의 넙치에 면역 작용에 대한 저항성 시험을 통해 이 병원체의 감염성에 대한 기초 자료를 얻고자 하였다.
시험균의 염분 농도별 발육 시험에서 강독성 균주 A28003가 다른 균주보다 광범위한 농도 조건에서 잘 성장하는 특성을 보였다. pH의 변화에 따른 시험균의 성장시험에서는 강독성 균주 A28003이 시험 pH 3.0~ 8.0의 범위 내에서 광범위하게 잘 성장하여 다른 시험 균주와 차이를 나타내었다. 시험어의 생체 방어 기구에 대한 시험균의 생존능은 체표 점액 생존능에서 A28003과 A19010 두 균주 모두 3시간 까지 그 수가 증가하였으나 3시간 이후 두 균주 모두 급격하게 감소하여 유의적인 차이를 볼 수 없었다. 시험균의 장 점액에 대한 생존능은 강독성 균주 A28003이 반응 직후 급격히 증가하여 약독성 균주에 비해 유의적인 차이가 나타내었다. 혈청에 대한 저항성 시험에서 두 균주 사이에는 반응 1시간 이후부터 유의적이 차이를 나타내어 강독성 균주의 저항성이 높은 것을 확인할 수 있었다. 각 시험 균주별 넙치 두신 leucocytes에서의 adhesion과 invasion 활성을 측정한 결과를 강독성 균주가 약독성 균주보다도 Adhesion능과 invasion능이 높은 것으로 나타났다. H2O2의 감수성 시험에서 강독성 균주와 약독성 균주 모두 H2O2의 영향을 받아 시간의 흐름에 따라 균수가 감소함을 알 수 있었다. 시험균을 인위 감염시킨 넙치 혈청 라이소자임 활성은 강독성 균주 시험구에서 약독성 시험구에 비해 유의적인 차이를 나타내었다. 시험어에 시험균을 인위 감염시켰을 때 혈청 내 AST 및 ALT 값이 강독성 균주 시험구에서 약독성 균주 시험구에 비하여 유의적인 차이가 나타났다.
이상의 시험 결과로부터 강독성 균주와 약독성 균주의 특성이 시험어의 생체 방어 기구 중 장점액과 혈청에 대해서 유의적인 생존능의 차이가 있음을 밝혔다. 따라서 이러한 차이가 병원성 발현의 영향을 주는 요소로 작용되었을 것으로 생각되며, 이외에도 넙치 두신의 leucocyte에 대한 Adhesion능과 invasion능의 차이를 비롯한 여러 가지 요소가 강독성 균주의 생존능을 높이는데 영향을 주어 약독성 균주보다 더 강한 병원성을 나타내게 되었을 것으로 사료된다.
- Author(s)
- 김민경
- Issued Date
- 2014
- Awarded Date
- 2014. 2
- Type
- Dissertation
- Publisher
- 부경대학교
- URI
- https://repository.pknu.ac.kr:8443/handle/2021.oak/1434
http://pknu.dcollection.net/jsp/common/DcLoOrgPer.jsp?sItemId=000001966843
- Affiliation
- 대학원
- Department
- 대학원 수산생명의학과
- Advisor
- 박수일
- Table Of Contents
- 목차
목차 ··············································································· ⅰ
Abstract ·········································································· ⅲ
Ⅰ. 서론 ··········································································· 1
Ⅱ. 재료 및 방법 ································································· 3
1. 시험어 ········································································ 3
2. 시험균주 ····································································· 3
3. V. scophthalmi 강독성 균주와 약독성 균주의 발육 성상 비교 ······ 5
3.1. 염분 농도에 따른 발육 시험 ·········································· 5
3.2. pH에 따른 발육 시험 ··················································· 5
4. 시험어의 면연능에 대한 시험균의 저항성 비교 ························ 6
4.1. 체표 점액 내에서의 생존능 · ··········································· 6
4.1.1. 체표 점액 분리 ····················································· 6
4.1.2. 체표 점액 내에서의 생존능 시험 ································· 6
4.2. 장 점액 내에서의 생존능 ·············································· 6
4.2.1. 장 점액 분리 ························································ 6
4.2.2. 장 점액 내에서의 생존능 시험 ··································· 7
4.3. 혈청 내에서의 생존능 시험 ············································ 7
4.3.1. 혈청 분리 ···························································· 7
4.3.2. 혈청에서의 생존능 시험 ··········································· 8
5. 시험균의 어류 두신 leucocyte에 대한 adhesion, invasion ············ 8
5.1. 넙치 두신 leucocyte 분리 ·············································· 8
5.2. Adhesion assay ························································· 9
5.3. Invasion assay ·························································· 9
6. H2O2 감수성 시험 ························································ 10
7. 시험어에 대한 공격 시험 ················································ 10
7.1. 인위 감염 시험 방법 ·················································· 10
7.2. 혈청 성분에 미치는 영향 ··············································11
7.3. 혈청 lysozyme activity에 미치는 영향 ···························11
8.시험균주에 대한 넙치 누적폐사율 ···········································12
Ⅲ. 결과 ···········································································13
1. 염분 농도 및 pH에 따른 발육정도 비교 ································13
2. 체표 점액에서의 생존능 ···················································15
3. 장 점액에서의 생존능 ·····················································16
4. 혈청 내에서의 생존능 ·····················································17
5. 시험균의 어류 두신 leucocyte에 대한 adhesion ·······················18
6. 시험균의 어류 두신 leucocyte에 대한 invasion ························19
7. H2O2 감수성 시험 ··························································20
8. AST 변화 ···································································21
9. ALT 변화 ···································································22
10. 혈청 lysozyme activity에 미치는 영향 ································23
11. 시험균주에 대한 넙치 누적폐사율 ····································24
Ⅳ. 고찰 ···········································································25
Ⅴ. 요약 ···········································································30
Ⅵ. 감사의 글 ·····································································32
Ⅶ. 참고 문헌 ·····································································34
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