JP4143530B2 - 魚類冷水病ワクチン - Google Patents
魚類冷水病ワクチン Download PDFInfo
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- JP4143530B2 JP4143530B2 JP2003417937A JP2003417937A JP4143530B2 JP 4143530 B2 JP4143530 B2 JP 4143530B2 JP 2003417937 A JP2003417937 A JP 2003417937A JP 2003417937 A JP2003417937 A JP 2003417937A JP 4143530 B2 JP4143530 B2 JP 4143530B2
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Description
(1)フラボバクテリウム サイクロフィラムG3724(以下の実験でもこの株を使用した。)の一白金耳を4mLのMCYT培地(トリプトン2.0g、酵母エキス0.5g、肉エキス0.2g、酢酸ナトリウム0.2g、塩化カルシウム0.2g、蒸留水1000mL、pH7.2)に接種し、15℃で2日間培養後、そのうちの0.5mLを200mL MCYT培地に接種し、15℃で振盪培養した。培養時間と菌体数及び600nmにおける光学密度との関係を図1に示す。図1から明らかなように、本菌は0〜24時間までが誘導期であり、24〜48時間までが対数増殖期であり、48時間以降が定常期であることがわかる。
フラボバクテリウム サイクロフィラムG3724を2000mL容坂口フラスコ中で1000mL MCYT培地中15℃で培養した。OD600nmが0.2〜0.7のものを対数増殖期菌体として用いた。すなわち、培養24〜36時間の間のOD600nmが0.2〜0.7の時期の培養物を、0.3%ホルマリン中で15℃、2日間インキュベーションして不活化菌体とし、次いで4℃で8,000〜10,000×gで遠心分離して不活化菌体とした。また、対照として培養36時間経過後(OD600nm=1.0)の培養物を同様に不活化して定常期不活化菌体を得た。
フラボバクテリウム サイクロフィラムG3724の一白金耳を50mLのMCYT培地に接種し、15℃で48時間予備培養した。このうち2.5mLを1000mLのMCYT培地に接種し、15℃で36時間培養した。このときOD600nmは0.2〜0.7であった。培養物を0.3%ホルマリン中で15℃、2日間インキュベーションした。次いで、4℃で8,000〜10,000×gで遠心分離して菌体を採取した。得られた菌体をさらに0.3%ホルマリン生理食塩水に再懸濁し、本菌の不活化菌体を含むワクチン組成物を得た。
平均体重5.0gのアユに、実施例2で得た対数増殖期及び定常期の菌体から得た不活化菌体を、0.1FKCg/kg/dayで経口投与した。
このように経口投与したアユに対して浸漬攻撃実験を行なった。その結果を表1に示す。
実施例3で得たワクチン組成物を用いてワクチン効果を検討した。すなわち、攻撃試験開始5週間前からワクチンを2週間経口投与(0.1g/kg)を行ない、その後、3週間免疫活性を上昇させる為に通常飼料で飼育を行なった。その後、3週間後に攻撃試験を実施する区と、ワクチン投与終了から、7週後に攻撃試験を実施する2区を設定し、攻撃試験を実施した。
0.5gのアユの2000尾に対して、ワクチンを毎日経口投与した区と、2週間で5回投与した(中2日で経口投与)区の2区を設けた。結果を表2、図7及び図8に示す。
さらに、ワクチン投与後、7週間で攻撃試験を実施した場合、ワクチンを投与した両区において対照区よりも有意にワクチン効果が高かった。
フラボバクテリウム サイクロフィラム NCMB1947をMCYT培地中15℃の振盪培養し、培養24〜48時間の間のOD600nmが0.2〜0.7の対数増殖期の培養液を人工感染に用いた。すなわち、対数増殖期の本菌を106〜108CFU/mLになるようにニジマスの水槽に添加し、浸漬方法による人工感染を試みた。なお、供試したニジマスは1〜4gであり、水温は15℃とした。その結果、図11に示すように対照群(非感染群)の死亡率が0%であったのに対して、対数増殖期の本菌感染群は死亡率が55.8%であり、本菌の浸漬方法によるニジマスに対する人工感染に初めて成功した。図12に健康なニジマス(A)、1日目に死亡したニジマスの症状(B、C、D)、5日目に死亡したニジマスの症状(E、F)及び死亡したニジマスの尾鰭から発見されたフラボバクテリウム サイクロフィラムを示す。
Claims (3)
- フラボバクテリウム サイクロフィラムの対数増殖期の不活化菌体を有効成分とする魚類冷水病ワクチン。
- フラボバクテリウム サイクロフィラムの対数増殖期の不活化菌体を含有する魚類冷水病ワクチン組成物。
- フラボバクテリウム サイクロフィラムの対数増殖期の不活化菌体の有効量を投与することを特徴とする魚類冷水病の予防法。
Priority Applications (1)
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JP2003417937A JP4143530B2 (ja) | 2002-12-18 | 2003-12-16 | 魚類冷水病ワクチン |
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JP2002366769 | 2002-12-18 | ||
JP2003417937A JP4143530B2 (ja) | 2002-12-18 | 2003-12-16 | 魚類冷水病ワクチン |
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JP2004210769A JP2004210769A (ja) | 2004-07-29 |
JP4143530B2 true JP4143530B2 (ja) | 2008-09-03 |
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Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
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JPS58102108A (ja) * | 1981-12-14 | 1983-06-17 | Mitsubishi Electric Corp | レ−ザ測距装置 |
JP6218512B2 (ja) * | 2013-09-03 | 2017-10-25 | 和歌山県 | アユ冷水病菌由来毒素 |
JPWO2018062527A1 (ja) * | 2016-09-29 | 2019-06-24 | Jx金属株式会社 | レーザー焼結用表面処理金属粉 |
JP2024033734A (ja) * | 2022-08-31 | 2024-03-13 | 滋賀県 | ワクチン製剤及びその製造方法、並びに魚類細菌感染症の予防方法 |
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