JP2006296390A

JP2006296390A - Freezing and storing liquid of fish fertilized egg and preparation method thereof

Info

Publication number: JP2006296390A
Application number: JP2005146003A
Authority: JP
Inventors: Kiyoyasu Murakami; 清康村上
Original assignee: Individual
Current assignee: Individual
Priority date: 2005-04-15
Filing date: 2005-04-15
Publication date: 2006-11-02

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a preservative that is used for freezing and storing fish sperm and fertilized egg for the purpose of species preservation and sea farming and releasing of horse mackerel, etc., especially a preservative that comprises a polyethylene glycol and mannitol active ingredients and a phosphate buffer solution and is used for freezing and storing live cells and an enzyme. <P>SOLUTION: The preservative stores fish fertilized egg for a long period of time and comprises 10%(W/V) polyethylene glycol, 1 mol (molecular weight W/V)mannitol and a phosphate buffer solution at pH 7.9. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

発明の詳細な説明Detailed Description of the Invention

［産業上の利用分野］
本発明は魚類、貝類等の***、卵、受精卵の生細胞、酵素を凍結状態または低温で長期間保存に必要な生細胞および酵素の保存に関するもの[Industrial application fields]
The present invention relates to storage of sperm such as fish and shellfish, eggs, live cells of fertilized eggs, live cells necessary for long-term storage of enzymes in a frozen state or at a low temperature, and enzymes.

［従来の技術］
特公昭６０−２９４７１号、特開平５−３８２８４号等今日まで多くの研究が行われているが、サカナ等の受精卵の凍結保存によるところの孵化は皆無である[Conventional technology]
Many researches have been conducted to date, such as Japanese Patent Publication No. 60-29471 and Japanese Patent Laid-Open No. 5-38284, but there has been no hatching due to cryopreservation of fertilized eggs such as fish.

［発明が解決しようとする課題］
魚類などの***、受精卵の凍結保存の目的は、種の保存の必要性ならびにクロマグロ等の養殖及び稚魚放流にある。[Problems to be solved by the invention]
The purpose of cryopreservation of sperm such as fish and fertilized eggs is the necessity of preserving the species and the cultivation of bluefin tuna and the release of fry.

［課題を解決するための手段］
１〜２日の受精卵を容器に入れ、凍結保存液を注入し、氷蔵庫に低温保管する。その後ドライアイス入りエチルアルコ−ル液で急速凍結し、氷点下８５℃の冷凍庫にて保管する。
［解凍方法］
解凍は２８〜３０℃の温度とする。[Means for solving problems]
Place the fertilized eggs for 1 to 2 days into a container, inject a cryopreservation solution, and store in a freezer. Thereafter, it is rapidly frozen with an ethyl alcohol solution containing dry ice and stored in a freezer at 85 ° C. below freezing.
[Thawing method]
Thawing is performed at a temperature of 28-30 ° C.

［作用］
魚の受精卵はそのままだと５〜１０時間で劣化すると言われている。
凍結保存液の作用は不明である。[Action]
It is said that the fertilized egg of a fish will deteriorate in 5 to 10 hours if left as it is.
The effect of the cryopreservation solution is unknown.

［実施例］
１〜２日の受精卵を２ｍｌ容器に入れ、凍結保存液を注入し、氷蔵庫にて低温保管する。
ドライアイス入りエチルアルコ−ル液で急速凍結し、氷点下８５℃にて１〜３ケ月冷凍庫に保管した。解凍を２８〜３０℃で行い孵化させた。[Example]
Place the fertilized egg for 1-2 days in a 2 ml container, inject a cryopreservation solution, and store at low temperature in an ice storage.
It was snap frozen with ethyl alcohol solution containing dry ice and stored in a freezer at 85 ° C. below freezing for 1 to 3 months. Thawing was performed at 28-30 ° C. and hatched.

［発明の効果］
凍結受精卵が２０〜３０％の割合で孵化した。[The invention's effect]
Frozen fertilized eggs hatched at a rate of 20-30%.

Claims

ポリエチレングリコ−ル、マンニト−ルの有効成分ならびにリン酸緩衝液の緩衝作用による生細胞および酵素の保存剤Active ingredients of polyethylene glycol and mannitol, and preservatives for living cells and enzymes by buffering action of phosphate buffer

１０％（Ｗ／Ｖ）ポリエチレングリコ−ル、
１モル（分子量Ｗ／Ｖ）マンニト−ル
ｐＨ７．９リン酸緩衝液
に調整されたことを有効とする請求項１記載の生細胞および酵素の保存剤10% (W / V) polyethylene glycol,
The preservative for living cells and enzymes according to claim 1, wherein the preservative is effectively adjusted to 1 mol (molecular weight W / V) mannitol pH 7.9 phosphate buffer.

***、卵、受精卵の生細胞および酵素の凍結保存、低温保存を有効とする請求項１、請求項２記載の生細胞および酵素の保存剤3. The viable cell and enzyme preservative according to claim 1, wherein cryopreservation and cryopreservation of viable cells and enzymes of sperm, eggs and fertilized eggs are effective.