JP5376455B2 - Crustacean ripening and / or egg-laying composition - Google Patents
Crustacean ripening and / or egg-laying composition Download PDFInfo
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Description
本発明は、甲殻類の催熟及び/又は産卵用組成物に関する。具体的には、本発明は、ゴカイ類抽出物から単離された中性脂質画分及び/又は遊離アミノ酸・ペプチド画分を含む、甲殻類、好ましくはクルマエビの催熟及び/又は産卵用組成物に関する。 The present invention relates to a crustacean ripening and / or egg-laying composition. Specifically, the present invention provides a composition for ripening and / or laying eggs of crustaceans, preferably prawns, comprising a neutral lipid fraction and / or a free amino acid / peptide fraction isolated from a coral extract. Related to things.
甲殻類の一種であるクルマエビは、重要な養殖種であるにも関わらず、天然から得られた親エビから卵を採って、稚エビに育てた後、養殖している。天然資源の減少や天然保菌エビの増加によって、稚エビの品質低下が懸念されている。そこで、人工飼育化で親エビを育成し、卵を確保する技術の開発が望まれている。したがって、親エビを催熟させ、産卵させるための高品質飼料の開発が急務である。 The shrimp, a kind of crustacean, is an important aquaculture species, but it is cultivated after collecting eggs from the parent shrimp obtained from nature and raising them to juvenile shrimp. Due to the decrease in natural resources and the increase in natural carrier shrimp, there is concern about the decline in the quality of juvenile shrimp. Therefore, it is desired to develop a technique for raising parent shrimp and securing eggs by artificial breeding. Therefore, there is an urgent need to develop a high quality feed for ripening and spawning shrimp.
現在までに、砂泥域でクルマエビの天然飼料となっていると考えられるイソゴカイがクルマエビの催熟効果を有すること、及び特にイソゴカイ抽出脂質がかかる催熟効果を有することが示唆されている(非特許文献1)。 To date, it has been suggested that isogokai, which is considered to be a natural feed for prawns in the sand and mud area, has a ripening effect on prawns, and that in particular, the extracted lipids of isopipe have such a ripening effect (non- Patent Document 1).
本発明は、甲殻類の催熟及び/又は産卵を効率的に促進させることができる新規な組成物を提供することを目的とする。 An object of the present invention is to provide a novel composition capable of efficiently promoting crustacean ripening and / or egg-laying.
本発明者らは、上記課題を解決するために、イソゴカイから抽出された各画分の催熟及び/又は産卵への効果を分析した結果、中性脂質を含む画分と遊離アミノ酸・ペプチドを含む画分が優れた甲殻類の催熟効果を有することを見出し、本発明を完成させるに至った。 In order to solve the above-mentioned problems, the present inventors analyzed the effect on the ripening and / or egg-laying of each fraction extracted from isogoyakai, and as a result, the fraction containing neutral lipid and the free amino acid peptide were obtained. It has been found that the contained fraction has an excellent crustacean ripening effect and has led to the completion of the present invention.
すなわち本発明は以下の特徴を包含する。
(1) ゴカイ類抽出物から単離された中性脂質画分及び/又は遊離アミノ酸・ペプチド画分を含み、ゴカイ類由来の前記中性脂質の含有量は乾燥重量で0.5重量％以上であり、ゴカイ類由来の前記遊離アミノ酸・ペプチドの含有量は乾燥重量0.1重量％以上である、甲殻類の催熟及び/又は産卵用組成物。
That is, the present invention includes the following features.
(1) A neutral lipid fraction and / or a free amino acid / peptide fraction isolated from a coral extract, wherein the content of the neutral lipid derived from coral is 0.5% by weight or more in dry weight A composition for ripening and / or laying eggs of a crustacean, wherein the content of the free amino acids / peptides derived from squirrels is 0.1% by weight or more in dry weight.
(2) ゴカイ類はイソゴカイである上記(1)記載の組成物。
(3) 甲殻類はクルマエビである上記(1)記載の組成物。
(2) The composition according to the above (1), wherein the shellfish is isopagokai.
(3) The composition according to (1) above, wherein the crustacean is prawns.
(4) 前記中性脂質画分は、ゴカイ類原料から1種以上の有機溶媒を用いて溶媒抽出されたものである、上記(1)記載の組成物。 (4) The composition according to the above (1), wherein the neutral lipid fraction is obtained by solvent extraction from a cormorant raw material using one or more organic solvents.
(5) 有機溶媒は、メタノール、クロロホルム及びアセトンである、上記(4)記載の組成物。
(6) 前記遊離アミノ酸・ペプチド画分は、ゴカイ類原料から1種以上の有機溶媒を用いて溶媒抽出されたものである、上記(1)記載の組成物。
(5) The composition according to (4) above, wherein the organic solvent is methanol, chloroform and acetone.
(6) The composition according to the above (1), wherein the free amino acid / peptide fraction is obtained by solvent extraction from a raw material of gokai using one or more organic solvents.
(7) 有機溶媒は、メタノール、クロロホルム及びトリクロロ酢酸である、上記(6)記載の組成物。
(8) 上記(1)〜(7)のいずれか記載の組成物を飼料に添加することを含む、甲殻類の催熟用飼料の製造方法。
(7) The composition according to (6) above, wherein the organic solvent is methanol, chloroform and trichloroacetic acid.
(8) A method for producing a crustacean ripening feed, comprising adding the composition according to any one of (1) to (7) above to the feed.
本発明によれば、甲殻類の催熟及び/又は産卵を効率的に促進させることができる組成物が提供される。 According to the present invention, a composition capable of efficiently promoting crustacean ripening and / or egg laying is provided.
本発明は甲殻類の催熟及び/又は産卵用組成物（以下、本発明の組成物と称する）に関する。本発明で使用する用語「催熟及び/又は産卵用」は、甲殻類に給餌することによって催熟、産卵を促進することを目的とする用途をいい、より具体的には、甲殻類の成熟の促進、卵影比の増大、産卵までに要する日数の短縮、産卵率の改善、卵質の改善、孵化率の向上などの1つ以上を目的とする用途を指す。本発明の組成物は、成熟前又は成熟後のいずれの甲殻類に対して給餌してもよいが、好ましくは成熟後の甲殻類に使用する。 The present invention relates to a crustacean ripening and / or egg-laying composition (hereinafter referred to as the composition of the present invention). The term “for ripening and / or egg laying” used in the present invention refers to an application aimed at promoting ripening and egg laying by feeding crustaceans, more specifically maturation of crustaceans. This is intended to be used for one or more of the following purposes: promotion of growth, increase of egg shadow ratio, reduction of days required for egg laying, improvement of egg production rate, improvement of egg quality, and improvement of hatching rate. The compositions of the present invention may be fed to either pre-mature or post-mature crustaceans, but are preferably used on post-mature crustaceans.
本発明の組成物の使用対象となる甲殻類は特に限定されず、例えば食用又は養殖用であるエビ類、カニ類、ザリガニ類、シャコ類、アミ類など広範な甲殻類が含まれる。エビ類として、これに限定されるものではないが、例えばクルマエビ、コウライエビ、ショウナンエビ、フトミゾエビ、テラオクルマエビ、テンジククルマエビ、ウシエビ、アカオエビ、クマエビ、ブルーシュリンプ、レッドテールシュリンプ、バナナシュリンプ、インドエビ、ホワイトシュリンプ（バナメイエビ）などのクルマエビ類、テナガエビ、ミナミテナガエビ、ヤマトテナガエビ、オニテナガエビ、コンジンテナガエビなどのテナガエビ類、およびイセエビ、ハコエビ、ゴシキエビ、ウチワエビ、セミエビなどのイセエビ類を挙げることができる。またカニ類としては、アサヒガニ、上海ガニ、ガザミ、タイワンガザミ、ノコギリガザミ、モクズガニ、ズワイガニ、ベニズワイガニ、ケガニ、タカアシガニなどを挙げることができ、ザリガニ類としてはアメリカンロブスター、ヨーロピアンロブスター、アカザエビ、アメリカザリガニ、ニホンザリガニなどを挙げることができる。 Crustaceans to be used for the composition of the present invention are not particularly limited, and include a wide range of crustaceans such as shrimps, crabs, crayfishes, clams, and mites that are edible or aquaculture. Examples of shrimp include, but are not limited to, shrimp, shrimp, shrimp, shrimp, terra prawn, tiger shrimp, bull shrimp, red shrimp, bear shrimp, blue shrimp, red tail shrimp, banana shrimp, indian shrimp, white shrimp Examples include prawns such as shrimp, shrimp, southern shrimp, shrimp, shrimp, shrimp, shrimp, shrimp, shrimp, shrimp, shrimp, etc. Examples of crabs include Asahi Crab, Shanghai Crab, Crab, Thai Crab, Saw Crab, Crab Crab, Snow Crab, Crab Crab, Crab Crab, Crab Crab, and Crayfish Crab Crab. Examples include crayfish.
本発明の組成物は、ゴカイ類由来の中性脂質及び/又は遊離アミノ酸・ペプチドを含んでおり、その含有量が、前者については、乾燥重量で0.5重量％以上、好ましくは0.6重量％以上、より好ましくは0.7重量％以上であり、後者については、乾燥重量で0.1重量％以上、好ましくは0.12重量％以上、より好ましくは0.14重量％以上であることを特徴とする。このような本発明の組成物は、ゴカイ類抽出物から中性脂質画分及び/又は遊離アミノ酸・ペプチド画分を単離することによって製造することができる。 The composition of the present invention contains neutral lipids and / or free amino acids / peptides derived from coral, and the content of the former is 0.5% by weight or more, preferably 0.6% by weight or more in terms of dry weight, More preferably, it is 0.7% by weight or more, and the latter is characterized by 0.1% by weight or more, preferably 0.12% by weight or more, more preferably 0.14% by weight or more in dry weight. Such a composition of the present invention can be produced by isolating a neutral lipid fraction and / or a free amino acid / peptide fraction from a coral extract.
本発明の組成物の原料となるゴカイ類は、分類学的にゴカイ類に分類されるものであれば特に制限されず、例えばイソゴカイ、アオゴカイなどを挙げることができる。特に本発明の組成物をクルマエビ類に給餌する場合には、原料としてイソゴカイを使用することが好ましい。またゴカイ類の形態は特に制限されず、生存形態であっても乾燥形態であってもよい。 There are no particular limitations on the coral species used as the raw material for the composition of the present invention, so long as they are classified taxonally as taxonomy, and examples thereof include isogokai and blue crab. In particular, when feeding the composition of the present invention to prawns, it is preferable to use isogokai as a raw material. In addition, the form of the spider is not particularly limited, and may be a living form or a dry form.
ゴカイ類抽出物からの中性脂質画分の単離は、ゴカイ類原料から脂溶性成分を抽出し、該脂溶性成分から中性脂質のみを単離する操作によって行うことができる。上記のような中性脂質画分の単離は、例えば1種以上の有機溶媒を用いた溶媒抽出処理によって行うことができる。かかる溶媒抽出処理に使用することができる有機溶媒としては、これに限定されるものではないが、メタノール、エタノール、クロロホルム、アセトン、ジエチルエーテルなどを挙げることができる。有機溶媒としてメタノール、クロロホルム及びアセトンを使用することが好ましい。 Isolation of the neutral lipid fraction from the coral extract can be performed by extracting a fat-soluble component from the coral material and isolating only the neutral lipid from the fat-soluble component. The neutral lipid fraction as described above can be isolated, for example, by solvent extraction using one or more organic solvents. Examples of the organic solvent that can be used for the solvent extraction treatment include, but are not limited to, methanol, ethanol, chloroform, acetone, diethyl ether, and the like. Methanol, chloroform and acetone are preferably used as the organic solvent.
有機溶媒としてメタノール、クロロホルム及びアセトンを使用する場合には、例えば、次のようにして中性脂質を単離することができる。すなわち、ホモジナイザー等により粉砕したゴカイ類原料にメタノール/クロロホルム混合液(2:1〜1：2、例えば1:1の割合)を添加して攪拌した後、濾過により残渣を除き、メタノール層とクロロホルム層とに分離するまで静置する。次いで、クロロホルム層を回収し、クロロホルムを除去する。次いで、5〜10倍量の冷アセトンを添加して静置し、上澄みを回収する操作を3〜5回繰り返す。このようにして回収された上澄みを全て合わせ、必要に応じて中性脂質を濃縮、乾燥処理に付すことにより、目的とする中性脂質画分を得ることができる。 When methanol, chloroform and acetone are used as the organic solvent, for example, neutral lipids can be isolated as follows. That is, a methanol / chloroform mixture (2: 1 to 1: 2, for example, a ratio of 1: 1) was added to a mulberry raw material pulverized with a homogenizer or the like, stirred, and then the residue was removed by filtration. The methanol layer and chloroform Let stand until separated into layers. Next, the chloroform layer is recovered and the chloroform is removed. Then, 5 to 10 times the amount of cold acetone is added and left to stand, and the operation of collecting the supernatant is repeated 3 to 5 times. By combining all the supernatants collected in this manner and concentrating and drying the neutral lipid as necessary, the objective neutral lipid fraction can be obtained.
ゴカイ類抽出物からの中性脂質の単離は上記手法に制限されず、当業者に公知の他の手法を用いて行ってもよい。そのような手法として、例えばブライとダイアーの方法(Can. J. Biochem. Physiol., 37:911-917, 1959参照)、フォルチの方法(J. Biol. Chem., 226:497-509, 1957参照)などを挙げることができる。 Isolation of the neutral lipid from the coral extract is not limited to the above method, and may be performed using other methods known to those skilled in the art. Examples of such methods include the Blye and Dyer method (see Can. J. Biochem. Physiol., 37: 911-917, 1959) and the Forch method (J. Biol. Chem., 226: 497-509, 1957). For example).
こうして単離される中性脂質画分は、中性脂質として主に、遊離脂肪酸、トリアシルグリセロール、ステロール類(例えばコレステロールなど)、トリ(及びモノ)アシルグリセロールを含んでいる(下記表2参照)。 The neutral lipid fraction thus isolated mainly contains free fatty acids, triacylglycerols, sterols (such as cholesterol), and tri (and mono) acylglycerols as neutral lipids (see Table 2 below). .
ゴカイ類抽出物からの遊離アミノ酸・ペプチド画分の単離は、ゴカイ類原料から水溶性成分を抽出し、該水溶性成分から遊離アミノ酸及びペプチドのみを単離する操作によって行うことができる。上記のような遊離アミノ酸及びペプチドの単離は、例えば1種以上の有機溶媒を用いた溶媒抽出処理によって行うことができる。遊離アミノ酸及びペプチドの単離に使用することができる有機溶媒としては、これに限定されるものではないが、メタノール、エタノール、クロロホルム、ジエチルエーテル、トリクロロ酢酸などを挙げることができる。有機溶媒としてメタノール、クロロホルム及びトリクロロ酢酸を使用することが好ましい。 Isolation of the free amino acid / peptide fraction from the coral extract can be performed by extracting a water-soluble component from the coral material and isolating only the free amino acid and the peptide from the water-soluble component. Isolation of free amino acids and peptides as described above can be performed, for example, by solvent extraction using one or more organic solvents. Examples of organic solvents that can be used for isolation of free amino acids and peptides include, but are not limited to, methanol, ethanol, chloroform, diethyl ether, trichloroacetic acid, and the like. Methanol, chloroform and trichloroacetic acid are preferably used as the organic solvent.
有機溶媒としてメタノール、クロロホルム及びトリクロロ酢酸を使用する場合には、例えば、次のようにして遊離アミノ酸・ペプチドを単離することができる。すなわち、ホモジナイザー等により粉砕したゴカイ類原料にメタノール/クロロホルム混合液(2:1〜1：2、例えば1:1の割合)を添加して攪拌した後、濾過により残渣を除き、メタノール層とクロロホルム層とに分離するまで静置する。次いで、メタノール層を回収し、メタノールを除去する。次いで、5〜10％濃度までトリクロロ酢酸を添加し、生成した沈殿を遠心分離により除去する。回収したトリクロロ酢酸可溶性画分は、その後必要に応じて、トリクロロ酢酸を除去するためのジエチルエーテルによる処理や当業者に公知の乾燥処理等に付し、目的とする遊離アミノ酸・ペプチドを得ることができる。 When methanol, chloroform and trichloroacetic acid are used as the organic solvent, for example, free amino acids / peptides can be isolated as follows. That is, a methanol / chloroform mixture (2: 1 to 1: 2, for example, a ratio of 1: 1) was added to a mulberry raw material pulverized with a homogenizer or the like, stirred, and then the residue was removed by filtration. The methanol layer and chloroform Let stand until separated into layers. The methanol layer is then collected and the methanol is removed. Then, trichloroacetic acid is added to a concentration of 5-10% and the resulting precipitate is removed by centrifugation. The recovered trichloroacetic acid-soluble fraction is then subjected to treatment with diethyl ether to remove trichloroacetic acid or drying treatment known to those skilled in the art to obtain the desired free amino acid / peptide, if necessary. it can.
ゴカイ類抽出物からの遊離アミノ酸・ペプチド画分の単離は上記手法に制限されず、当業者に公知の他の手法を用いて行ってもよい。そのような手法として、例えば限外ろ過法などを挙げることができる。 Isolation of the free amino acid / peptide fraction from the coral extract is not limited to the above method, and may be performed using other methods known to those skilled in the art. Examples of such a method include an ultrafiltration method.
本発明の組成物は、好ましくは、ゴカイ類抽出物に含まれる中性脂質と遊離アミノ酸・ペプチドの両方を含んでいる。下記実施例に示されるように、ゴカイ類抽出物に含まれる中性脂質及び遊離アミノ酸・ペプチドは、共に催熟及び産卵を促進する効果を有するため、両者を併用することによって前記効果を増強することができる。なおこの場合、本発明の組成物中の中性脂質画分と遊離アミノ酸・ペプチド画分の含有比は特に制限されない。 The composition of the present invention preferably contains both neutral lipids and free amino acids / peptides contained in the coral extracts. As shown in the examples below, neutral lipids and free amino acids / peptides contained in the coral extracts have the effect of promoting ripening and egg laying, and thus enhance the effect by using both in combination. be able to. In this case, the content ratio of the neutral lipid fraction and the free amino acid / peptide fraction in the composition of the present invention is not particularly limited.
上記の通り、ゴカイ類抽出物中に含まれる中性脂質及び/又は遊離アミノ酸・ペプチドは、甲殻類に摂食させることにより、甲殻類の成熟の促進、卵影比の増大、産卵までに要する日数の短縮、産卵率の改善、卵質の改善、孵化率の向上などの催熟、産卵を促進する効果を有するものである。したがって、本発明の組成物は、甲殻類の催熟、産卵用の飼料の製造に使用することができる。 As described above, neutral lipids and / or free amino acids / peptides contained in coral extracts are required to promote crustacean maturation, increase egg shadow ratio, and lay eggs by feeding them to crustaceans. It has effects of promoting ripening and egg-laying such as shortening the number of days, improving the egg-laying rate, improving the egg quality, and improving the hatching rate. Therefore, the composition of the present invention can be used for the ripening of crustaceans and the production of eggs for laying eggs.
本発明の甲殻類の催熟及び/又は産卵用飼料の製造方法は、本発明の組成物を各種飼料に配合することを含んでいる。本発明の方法に使用することができる飼料は、甲殻類への給餌に使用することができるものであれば特に制限されず、市販の人工飼料や天然餌料に配合させることができる。またその配合量も、催熟及び/又は産卵を効果的に促進できる量であれば特に制限されず、例えば飼料に対して、中性脂質の含有量が0.1〜1.0重量％、好ましくは0.2〜0.8重量％、例えば0.5重量％となるように上記中性脂質を配合させればよく、及び/又は遊離アミノ酸・ペプチドの含有量が0.02〜0.2重量％、好ましくは0.04〜0.16重量％、例えば0.1重量％となるように上記遊離アミノ酸・ペプチド画分を配合させればよい。 The method for producing a shellfish ripening and / or egg-laying feed of the present invention includes blending the composition of the present invention with various feeds. The feed that can be used in the method of the present invention is not particularly limited as long as it can be used for feeding shellfish, and can be blended with a commercially available artificial feed or natural feed. Further, the amount thereof is not particularly limited as long as it is an amount that can effectively promote ripening and / or egg laying, and for example, the content of neutral lipid is 0.1 to 1.0% by weight, preferably 0.2 to The neutral lipid may be added so as to be 0.8% by weight, for example 0.5% by weight, and / or the content of free amino acid / peptide is 0.02 to 0.2% by weight, preferably 0.04 to 0.16% by weight, for example 0.1 What is necessary is just to mix | blend the said free amino acid and peptide fraction so that it may become weight%.
以下、実施例により本発明をより詳細に説明するが、本発明は下記実施例に限定されるものではない。 EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited to the following Example.
本実施例では、養殖イソゴカイ（高知産）を使用した。イソゴカイ抽出物の作製に使用した溶媒（クロロホルム、メタノール、トリクロロ酢酸）は、ナカライテスク社製を用いた。 In this example, a cultured isogocai (Kochi product) was used. The solvent (chloroform, methanol, trichloroacetic acid) used for preparation of the isogokai extract was manufactured by Nacalai Tesque.
養殖イソゴカイは、入手後すぐに冷凍庫（−20℃）で凍結し、凍結乾燥機を用いて凍結乾燥した。イソゴカイ粉末100gに対して、メタノール250ml及びクロロホルム250mlを加え、カップホモジナイザーでよく混合し定性用濾紙（アドバンテック、No.3 ）を用いて、吸引濾過を行った。濾紙状の固形物（タンパク質、繊維等を含む）は、乾燥して冷凍保存した。濾液は、分液漏斗に移し、水約200mlを加え攪拌後、2層に分離するのを待ち、上層A（水溶性画分）と下層B（脂溶性画分）に分けた。 The cultured isagokai was frozen immediately after acquisition in a freezer (−20 ° C.) and freeze-dried using a freeze dryer. To 100 g of isogokai powder, 250 ml of methanol and 250 ml of chloroform were added, mixed well with a cup homogenizer, and subjected to suction filtration using a qualitative filter paper (Advantech, No. 3). The filter paper-like solid (including protein, fiber, etc.) was dried and stored frozen. The filtrate was transferred to a separatory funnel, and after adding about 200 ml of water and stirring, it was waited for separation into two layers, and divided into an upper layer A (water-soluble fraction) and a lower layer B (lipid-soluble fraction).
上層A（水溶性画分）は、ロータリーエバポレーターを用いて溶液中のメタノールを蒸発させたのち、水溶液の容積を測定し、トリクロロ酢酸（TCA）濃度が10％になるようTCAを添加した。溶液を冷蔵庫（4℃）で保存し、生成した沈殿と溶液を遠心分離によって、分離した。遠心分離の沈殿は水洗後、凍結乾燥して冷凍保存した。上澄みは、分液漏斗に移し、等量のジエチルエーテルを加え振とうした。静置し、二層分離後、溶液の上層を除去した。下層のpH が5〜6になるまで操作を繰り返し、溶液中のTCAを取り除いた。溶液は、ロータリーエバポレーターを用いて濃縮し、その後凍結乾燥して、遊離アミノ酸・ペプチド画分とし、凍結保存した。遊離アミノ酸・ペプチド画分の組成を下記表1に示す。 For the upper layer A (water-soluble fraction), the methanol in the solution was evaporated using a rotary evaporator, the volume of the aqueous solution was measured, and TCA was added so that the trichloroacetic acid (TCA) concentration was 10%. The solution was stored in a refrigerator (4 ° C.), and the produced precipitate and the solution were separated by centrifugation. The centrifugal precipitate was washed with water, freeze-dried and stored frozen. The supernatant was transferred to a separatory funnel, and an equal amount of diethyl ether was added and shaken. The mixture was allowed to stand, and after two-layer separation, the upper layer of the solution was removed. The operation was repeated until the pH of the lower layer reached 5-6, and TCA in the solution was removed. The solution was concentrated using a rotary evaporator and then freeze-dried to obtain a free amino acid / peptide fraction and stored frozen. The composition of the free amino acid / peptide fraction is shown in Table 1 below.
下層B（脂溶性画分）は、ロータリーエバポレーターを用いて溶媒を除去し、10倍量の冷アセトンを加え撹拌後、冷蔵保存した。1日静置後、上澄(中性脂質画分)と沈殿(極性脂質画分)に分離した。沈殿には冷アセトンをさらに加え、撹拌後、静置分離した。操作は上澄が無色になるまで繰り返し、上澄は全て合わせて、減圧濃縮し、中性脂質画分を得た。中性脂質画分の組成を下記表2に示す。 For the lower layer B (lipid-soluble fraction), the solvent was removed using a rotary evaporator, and 10 times the amount of cold acetone was added and stirred, and then stored refrigerated. After standing for 1 day, it was separated into a supernatant (neutral lipid fraction) and a precipitate (polar lipid fraction). Cold acetone was further added to the precipitate, and after stirring, the mixture was left to stand. The operation was repeated until the supernatant became colorless. All the supernatants were combined and concentrated under reduced pressure to obtain a neutral lipid fraction. The composition of the neutral lipid fraction is shown in Table 2 below.
沈殿は減圧乾燥し、極性脂質を得た。なお、本実施例で採用したゴカイ類抽出物の抽出フローチャートを図１に示す。 The precipitate was dried under reduced pressure to obtain polar lipids. In addition, FIG. 1 shows an extraction flow chart of the squirrel extract employed in this example.
実施例に用いた飼料は、イカ粉末、魚粉、オキアミ粉末及びイカ肝油を主体とし、コレステロール、ミネラル混合物、ビタミン混合物等を添加した。ゴカイ粉末もしくはゴカイ抽出画分をそれぞれ飼料に添加した。試験区を下記に示す：
試験区1：基本配合飼料
試験区2：基本配合飼料＋遊離アミノ酸・ペプチド画分0.5重量％添加
試験区3：基本配合飼料＋極性脂質画分0.5重量％添加
試験区4：基本配合飼料＋中性脂質画分0.5重量％添加
試験区5：冷凍ゴカイ
The feed used in the examples mainly consisted of squid powder, fish meal, krill powder and squid liver oil, and added with cholesterol, mineral mixture, vitamin mixture and the like. The flounder powder or the flounder extract fraction was added to the feed, respectively. The test areas are shown below:
Test plot 1: Basic formula feed Test plot 2: Basic formula feed + free amino acid / peptide fraction 0.5 wt% added Test plot 3: Basic formula feed + polar lipid fraction 0.5 wt% added Test plot 4: Basic formula feed + medium Test lipid 5
成熟試験には、松本水産株式会社（大分県）より入手した雌クルマエビを使用した。水槽収容時に、雌エビは、体重、体幅長および卵影幅を測定し、外科用縫合糸を用いて片眼柄処理を行い、個体識別できるようタグを取り付けた。成熟指標には卵影比を用いた。算出式は次のとおりである：
卵影比(%)=(卵影幅/体幅)×100
試験飼料を1日1回、15日間給餌し、2日おきに卵影幅長と体幅長を測定した。
For the maturation test, female prawns obtained from Matsumoto Suisan Co., Ltd. (Oita Prefecture) were used. When the aquarium was housed, the female shrimp measured the body weight, body width length, and egg shadow width, performed a one-eye pattern using a surgical suture, and attached a tag so that the individual could be identified. The egg shadow ratio was used as a maturity index. The formula is as follows:
Egg shadow ratio (%) = (Egg shadow width / body width) x 100
The test feed was fed once a day for 15 days, and the egg shadow width and body width were measured every two days.
図2に示すように、いずれの試験区でも実験開始時に比べ卵影比は増加していたが、遊離アミノ酸・ペプチド画分を添加した試験区2が最も良好な増加を示した。また、最大卵影比はほぼ同等の値を示した（図3）。 As shown in FIG. 2, the egg shade ratio increased in all the test groups compared to the start of the experiment, but the test group 2 to which the free amino acid / peptide fraction was added showed the best increase. In addition, the maximum egg shadow ratio showed almost the same value (Figure 3).
しかし、最大卵影比に達するまでの日数は、冷凍ゴカイ区が最も低い値を示したが、ゴカイ中性脂質添加区との間に有意差はみられず、ゴカイ中性脂質のクルマエビ卵巣成熟に対する効果が確認された(図4)。 However, the number of days to reach the maximum egg shadow ratio was the lowest in the frozen scallop group, but there was no significant difference from the scallop neutral lipid addition group. The effect on was confirmed (Fig. 4).
次に、交尾栓をもった雌クルマエビに、上記遊離アミノ酸・ペプチド画分(0.5重量％)及び中性脂質画分(0.5重量％)を添加した配合飼料を給餌し、その産卵に及ぼす効果を活ゴカイ給餌区と比較した。両区で産卵が観察されたが、活ゴカイ給餌区が若干高い産卵率(産卵個体/総個体数x100)を示した。また、産卵個体当たりの産卵数に差はみられなかった。しかし、孵化率(孵化数/産卵数x100)は、配合飼料給餌区が活ゴカイ給餌区に比べ有意に高く、1尾当たりの幼生数も配合飼料給餌区が高い値を示した(図5及び6)。 Next, the female prawns with mating plugs were fed with a mixed feed supplemented with the above free amino acid / peptide fraction (0.5 wt%) and neutral lipid fraction (0.5 wt%), and the effects on egg laying were examined. Compared with the live feeding area. Oviposition was observed in both plots, but the live feeding group showed a slightly higher egg-laying rate (spawning individuals / total number of individuals x100). Moreover, there was no difference in the number of eggs laid per egg laying individual. However, the hatching rate (number of hatched eggs / number of eggs laid x100) was significantly higher in the mixed feed feeding group than in the live feeding group, and the number of larvae per fish was also higher in the mixed feed feeding group (Figs. 5 and 5). 6).
以上の結果から、ゴカイ中性脂質画分又は遊離アミノ酸・ペプチド画分の添加により、クルマエビ催熟用飼料が作製可能であることが示された。 From the above results, it was shown that a prawn ripening feed can be prepared by adding a neutral lipid fraction or a free amino acid / peptide fraction.
本発明により、甲殻類の催熟及び/又は産卵を効率的に促進させることができる組成物が提供される。従って、本発明に係る組成物は、クルマエビ類などの甲殻類養殖などの分野において有用である。 The present invention provides a composition capable of efficiently promoting crustacean ripening and / or egg-laying. Therefore, the composition according to the present invention is useful in the field of crustacean culture such as prawns.
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