JP3302123B2 - Animal Plankton Culture Feed - Google Patents

Animal Plankton Culture Feed

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Publication number
JP3302123B2
JP3302123B2 JP24885193A JP24885193A JP3302123B2 JP 3302123 B2 JP3302123 B2 JP 3302123B2 JP 24885193 A JP24885193 A JP 24885193A JP 24885193 A JP24885193 A JP 24885193A JP 3302123 B2 JP3302123 B2 JP 3302123B2
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JP
Japan
Prior art keywords
weight
feed
algae
dha
zooplankton
Prior art date
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JP24885193A
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Japanese (ja)
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JPH0775507A (en
Inventor
清 長谷川
美代子 佐野
真 柏倉
Original Assignee
日清製油株式会社
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Description

【発明の詳細な説明】DETAILED DESCRIPTION OF THE INVENTION

【0001】[0001]

【産業上の利用分野】本発明は、栽培漁業・養殖におけ
る水産種苗の生産において必要不可欠なシオミズツボワ
ムシ、アルテミア等の動物性プランクトンを培養するた
めの飼料に関する。さらに詳しくは、栄養面でドコサヘ
キサエン酸を強化できる動物性プランクトン培養用飼料
に関する。
BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a feed for culturing zooplankton such as Rotifer, Artemia and the like, which are indispensable for the production of aquatic seedlings in cultivation, fishery and aquaculture. More specifically, the present invention relates to a feed for culturing zooplankton that can enhance docosahexaenoic acid in nutritional aspects.

【0002】[0002]

【従来の技術】水産分野の魚類および甲殻類の種苗生産
すなわち稚仔魚介類(以下、単に稚仔魚という)の生産
において、シオミズツボワムシ、アルテミア等の動物性
プランクトンは生物餌料として有用であり、必要不可欠
のものである。そして稚仔魚にとっては、エイコサペン
タエン酸(以下、EPAと略す)に代表されるω−3系
高度不飽和脂肪酸が必須脂肪酸であることが知られてい
る。しかるに稚仔魚の餌料となる動物性プランクトン自
体はω−3系高度不飽和脂肪酸をほとんど含有しない。
2. Description of the Related Art In the production of fish and crustaceans in the field of fisheries, i.e., the production of juveniles and larvae (hereinafter referred to simply as juveniles), zooplankton such as rotifers and artemia are useful as biological feeds. Is essential. For larvae, it is known that ω-3 highly unsaturated fatty acids represented by eicosapentaenoic acid (hereinafter abbreviated as EPA) are essential fatty acids. However, zooplankton, which is a feed for larvae, contains almost no ω-3 highly unsaturated fatty acids.

【0003】このため動物性プランクトンにω−3系高
度不飽和脂肪酸を強化し、その生体内に蓄積せしめる試
みがこれまでに行われてきた。例えば、海産クロレラと
通称され広く知られているナンノクロロプシス(Nannoc
hloropsis )属の藻類は、その藻体中にEPAを含み、
シオミズツボワムシやアルテミアを培養する際の好適な
飼料として利用されている。そしてこれを用いて培養し
た動物性プランクトンはEPAが強化され、稚仔魚の生
産においては成長率の向上、死亡率の低下等の点で効果
を発揮している。(「油脂」、第42巻、NO. 12、第
242〜246頁、1989年等)
[0003] For this reason, attempts have been made to enhance ω-3 polyunsaturated fatty acids in zooplankton and accumulate them in the body. For example, Nannochloropsis (Nannoc chloropsis), commonly known as marine chlorella
algae of the genus hloropsis) contain EPA in their algal bodies,
It is used as a suitable feed when cultivating rotifers and artemia. The zooplankton cultured using this has enhanced EPA and is effective in improving the growth rate and decreasing the mortality in the production of larvae and larvae. ("Oil", Vol. 42, No. 12, pp. 242-246, 1989, etc.)

【0004】ところで、近年、マダイやシマアジ等の海
産稚仔魚に対する必須脂肪酸としての効果は、EPAよ
りもドコサヘキサエン酸(以下、DHAと略す)の方が
大きいことが報告されている。(平成元年度日本水産学
会春季大会講演要旨集、第39頁および平成2年度日本
水産学会春季大会講演要旨集、第44頁)すなわち、人
工配合飼料中のEPAおよびDHA含量を変えて飼育し
た稚仔魚において、DHA添加区に成長促進効果を認め
ている。
[0004] In recent years, it has been reported that docosahexaenoic acid (hereinafter abbreviated as DHA) is more effective as an essential fatty acid for marine juveniles such as red sea bream and red horse mackerel than EPA. (Proceedings of the Spring Meeting of the Fisheries Society of Japan, pp. 39, and Abstracts of the Spring Meeting of the Fisheries Society of Japan, pp. 44), namely, seedlings reared by changing the EPA and DHA content in artificial feed In the larvae, the DHA-added group has a growth promoting effect.

【0005】したがって、稚仔魚を飼育するための動物
性プランクトンはDHAを含有するものが好ましく、こ
れを用いれば、稚仔魚に必須脂肪酸としてのDHAを容
易に摂餌させることができるのみならず、油分をはじめ
とする飼料成分が水中で分散し、水底にたい積して汚染
源となり、ひいては稚仔魚の生育を阻害することにもな
りかねない人工配合飼料に比べて、飼育効率の良い生物
餌料となる可能性がある。
[0005] Therefore, zooplankton for breeding larvae and larvae preferably contains DHA, which not only allows the larvae to easily feed DHA as an essential fatty acid, Oil and other feed components disperse in the water, deposit on the bottom of the water and become a source of contamination, resulting in a more efficient biological feed compared to artificial formula feed, which may inhibit the growth of larvae and larvae. there is a possibility.

【0006】そこでDHAを産生する藻類についてみて
みると、例えばJ.Protozoal 、第117巻(2)、第2
13〜219頁、1970年には、アムフィディニウム
(Amphidinium )属等のDHA含有藻類が報告されてい
る。また特開平4−346760号公報では、DHA含
有藻類としてのイソクリシス(Isochrysis)属の藻体を
動物プランクトン用飼料として利用することが開示され
ている。しかしながら、これらの藻類はいずれもその培
養において光の照射を必須条件とする独立栄養で生育す
るものであり、かかる藻類を動物プランクトン用飼料と
して用いようとすれば製造コストが極めて高価になり、
実用的でない。
[0006] When looking at algae that produce DHA, for example, J. Protozoal, Vol. 117 (2), Vol.
On pages 13 to 219, 1970, DHA-containing algae such as genus Amphidinium are reported. JP-A-4-346760 discloses that alga bodies of the genus Isochrysis as DHA-containing algae are used as feed for zooplankton. However, these algae are all grown under autotrophic conditions that require light irradiation in their culture.If such algae are used as feed for zooplankton, the production cost becomes extremely expensive,
Impractical.

【0007】これに対し、従属栄養で生育するDHA産
生藻類として、Phytochemistry、第27巻(6)、第1
679〜1683頁、1988年にはクリプセコディニ
ウム(Crypthecodinium )属の藻類が記載されている。
この先行文献によれば、クリプセコディニウム コーニ
ー(Crypthecodinium cohnii)は、その培養藻体中の脂
質のうち約72%がトリアシルグリセリドを主体とする
中性脂質であり、約28%がホスファチジルコリン等の
リン脂質を主体とする極性脂質である。そして総脂質の
構成脂肪酸のうち8.8%がDHAである。
On the other hand, PHATOCHEMISTRY, Vol. 27 (6), No. 1
679-1683, 1988, describe algae of the genus Crypthecodinium.
According to this prior art document, Crypthecodinium cohnii (Crypthecodinium cohnii) is a neutral lipid composed mainly of triacylglyceride in about 72% of lipids in the cultured algal cells, and about 28% is phosphatidylcholine or the like. Is a polar lipid mainly composed of phospholipids. And 8.8% of the constituent fatty acids of the total lipids is DHA.

【0008】しかしながら、かかる従属栄養で生育する
DHA産生藻類の産業的用途開発の検討はこれまでほと
んどなされていなかった。また動物性プランクトンを培
養してDHAを栄養強化するためにDHAを多量に含有
する藻類が有効であることは前記のとおり公知である
が、動物性プランクトンを効率よく培養するためのその
脂質形態については知られていなかった。
[0008] However, there has been little research on industrial use development of DHA-producing algae growing on such heterotrophs. Although it is known that an algae containing a large amount of DHA is effective for culturing zooplankton and fortifying DHA as described above, it is known that an alga containing DHA is effective for culturing zooplankton efficiently. Was not known.

【0009】[0009]

【発明が解決しようとする課題】かかる現状に鑑み、本
発明者らは、水産種苗の生産において稚仔魚用生物餌料
として有用な動物性プランクトンを培養するための飼料
について鋭意検討を行い、本発明を完成するに至った。
すなわち本発明の目的は、前期動物性プランクトンに対
して稚仔魚の必須脂肪酸の1種であるDHAを栄養強化
でき、前記動物性プランクトンの生産効率が良い、安価
な飼料を提供することにある。
In view of the above situation, the present inventors have conducted intensive studies on a feed for culturing zooplankton, which is useful as a biological feed for larvae and larvae in the production of marine seeds and seedlings. Was completed.
That is, an object of the present invention is to provide an inexpensive feed that can fortify DHA, which is one of the essential fatty acids of larvae, to early zooplankton, and that has high production efficiency of the zooplankton.

【0010】[0010]

【課題を解決するための手段】上記目的に対して、本発
明は、従属栄養で生育し、DHA含有極性脂質を含み、
かつ総脂肪酸中のDHA含量が10重量%以上である藻
類を原料としてなる動物性プランクトン培養用飼料、を
その要旨とする。まず、本発明において対象とする動物
性プランクトンとは、稚仔魚の生物餌料となるものであ
ればよく、具体的にはシオミズツボワムシ、アルテミ
ア、ミジンコ等を例示できる。
SUMMARY OF THE INVENTION To this end, the present invention is directed to a heterotrophic growing, comprising DHA-containing polar lipid,
The gist of the present invention is a feed for cultivating animal plankton, which is made from algae having a DHA content of 10% by weight or more in total fatty acids. First, the zooplankton to be used in the present invention may be any substance that can be used as a biological feed for fry and larvae, and specific examples include rotifers, artemia and daphnia.

【0011】次に本発明の飼料に係わる藻類について説
明する。本発明で用いる藻類は、従属栄養で培養が可能
な、DHA産生能のある藻類であれば、特にその種ある
いは株などを限定するものではない。この条件に該当す
る藻類としては、例えば「微生物学辞典1992年度
版」(技報堂出版)によれば、有色藻門、渦鞭毛藻綱、
ペリディニウム目、クリプセコディニウム属に属するク
リプセコディニウム コーニー(Crypthecodinium cohn
ii)、また褐色植物門、二毛菌綱、ミズカビ目のトラウ
ストキトリウム アウレウム(Thraustochytrium aureu
m )等があげられる。これらは American Type Culture
Collection(米国、略称ATCC)等の藻類研究機関
より容易に入手でき、前者の例としてATCC3002
1、同30334、同30336、同50052等、ま
た後者の例としてATCC34304、同28211等
がある。
Next, algae related to the feed of the present invention will be described. The alga used in the present invention is not particularly limited as long as it is an algae capable of culturing with heterotrophic nutrition and capable of producing DHA. Examples of the algae that meet this condition include, according to “Microbiological Dictionary 1992 Edition” (published by Gihodo), Colored Algae, Dinoflagellate,
Crypthecodinium cohn, belonging to the genus Crypsecodinium
ii) Also, brown plant phylum, dichomycete, Thraustochytrium aureu (Thraustochytrium aureu)
m) and the like. These are American Type Culture
Collection (US, abbreviated as ATCC) and other algal research institutions.
1, 30334, 30336, 50052, and the like, and examples of the latter are ATCC 34304, 28211, and the like.

【0012】本発明の飼料の原料として適する藻類は、
例えば前記藻類を培養したものであるが、その藻体中に
蓄積される脂質としてDHAを含有するリン脂質、糖脂
質等の極性脂質を含み、かつ脂質を構成する総脂肪酸の
うちDHAが10重量%以上を占めることを必須とす
る。動物性プランクトンを培養するための飼料原料とす
る藻類は、DHAをトリアシルグリセリド等の中性脂質
よりむしろ前記極性脂質の形態で藻体中に蓄積すること
が重要であり、かかるDHA含有極性脂質の全脂質中の
含有量は30重量%以上であることが望ましい。また藻
体中の脂質の総脂肪酸のうちDHA含有量が10重量%
以上であることを必要とし、好ましくは30重量%以上
である。10重量%未満であると、動物性プランクトン
にDHAを強化しにくくなる。
Algae suitable as a raw material for the feed of the present invention include:
For example, a culture of the algae, which contains polar lipids such as phospholipids and glycolipids containing DHA as lipids accumulated in the algae, and DHA is 10% by weight of the total fatty acids constituting the lipids It is mandatory to occupy more than%. It is important that algae used as feed materials for culturing zooplankton accumulate DHA in algal bodies in the form of the polar lipids rather than neutral lipids such as triacylglycerides. Is preferably 30% by weight or more in the total lipids. The DHA content is 10% by weight of the total fatty acids of the lipids in the alga body.
Or more, and preferably 30% by weight or more. If the amount is less than 10% by weight, it becomes difficult to enhance DHA in zooplankton.

【0013】本発明に係わる藻類を培養するには、次の
ように行なえばよい。すなわち食塩濃度が12〜25重
量%である天然もしくは人工海水培地、あるいはこれに
グルコース、ショ糖等の炭素源、アンモニウム塩、酵母
エキス、アミノ酸またはその塩等の窒素源、各種ビタミ
ン類およびミネラル類等を含む培地を用い、pHを5〜
7とし、20〜40℃で2〜10日間、通気培養する。
培養終了時期は必要とする藻体量が確保される任意の時
期でよいが、藻体細胞径は対数増殖中期前後では5〜1
5μm、定常期では10〜25μmとなり、これを適用
する動物性プランクトンの大きさに応じて培養を停止さ
せてもよい。
The cultivation of the algae according to the present invention may be carried out as follows. That is, a natural or artificial seawater medium having a salt concentration of 12 to 25% by weight, or a carbon source such as glucose or sucrose, a nitrogen source such as ammonium salt, yeast extract, amino acid or its salt, various vitamins and minerals Using a medium containing pH
The culture was aerated at 20 to 40 ° C for 2 to 10 days.
The cultivation end time may be any time at which the required amount of algal cells is secured, but the algal cell diameter is 5 to 1 around mid-logarithmic growth.
5 μm and 10 to 25 μm in the stationary phase, and the culture may be stopped according to the size of the zooplankton to which this is applied.

【0014】なお前記培養にあたり、藻類の増殖曲線に
おける指数増殖中期以降の任意の培養時期、好ましくは
対数増殖期から定常期に至る時期に、培地中の食塩濃度
を当該藻類の生育に好適な食塩濃度すなわち12〜25
重量%より0.1〜10重量%、好ましくは0.1〜3
重量%高い値に設定し、その食塩濃度で培養をさらに3
〜50時間程度行うことにより、DHA含有極性脂質が
30重量%以上、より実用的には50〜65重量%程
度、また総脂肪酸中のDHA含量が30重量%以上、よ
り実用的には45〜50重量%程度の藻体を得ることが
できる。
In the culturing, the salt concentration in the medium is adjusted at any time after the middle exponential growth period in the growth curve of the algae, preferably during the period from the logarithmic growth period to the stationary phase, by changing the salt concentration suitable for the growth of the algae. Concentration, ie 12-25
0.1 to 10% by weight, preferably 0.1 to 3% by weight
Weight% higher, and further culture at that salt concentration for 3 more days.
By carrying out for about 50 hours, the DHA-containing polar lipid is 30% by weight or more, more practically about 50 to 65% by weight, and the DHA content in the total fatty acids is 30% by weight or more, more practically 45 to 45%. Algae of about 50% by weight can be obtained.

【0015】このようにして得られる藻体は、培養終了
後、遠心分離、精密ないし限外濾過等の適当な手段で濃
縮し、要すれば水洗、減圧または噴霧乾燥、細胞破砕等
の処理を施し、これを単独もしくは他の公知の餌料成分
と混合して前記の動物性プランクトンの飼料とする。か
かる飼料で好ましくは12〜100時間培養し養成した
動物性プランクトンや2次培養をした動物性プランクト
ンは活力がよく、その総脂肪酸中のDHA含量は強化さ
れて10〜30重量%となる。
After completion of the cultivation, the alga thus obtained is concentrated by a suitable means such as centrifugation, precision or ultrafiltration, and if necessary, washed, washed under reduced pressure or spray dried, and crushed. This is used alone or mixed with other known feed components to obtain the above-mentioned zooplankton feed. The zooplankton cultivated and cultivated with such a feed for 12 to 100 hours or the zooplankton cultivated in the secondary culture is highly active, and the DHA content in the total fatty acids is enhanced to 10 to 30% by weight.

【0016】[0016]

【実施例】【Example】

参考例1 クリプセコディニウム コーニー(ATCC3033
6)を、表1に示した組成の培地3リットルに植えつけ
30℃にて、pHを6.8に調整しながらジャーファー
メンターで通気培養した。培養中は定期的にサンプリン
グし、O.D.660nmの濁度を測定した。その濁度
より増殖曲線を描き、対数増殖中期(85時間目)に培
養藻体を遠心分離して集め、さらに凍結乾燥して乾燥藻
体を32g得た。
Reference Example 1 Crypsecodinium coney (ATCC3033
6) was inoculated into 3 liters of a medium having the composition shown in Table 1, and aerated with a jar fermenter at 30 ° C. while adjusting the pH to 6.8. During the culture, samples were taken periodically. D. The turbidity at 660 nm was measured. A growth curve was drawn from the turbidity, and the cultured algal cells were collected by centrifugation at the middle stage of logarithmic growth (85 hours), and freeze-dried to obtain 32 g of dried algal cells.

【0017】[0017]

【表1】 [Table 1]

【0018】 *ビタミンミックス水溶液 ビオチン : 0.003g/リットル−溶液 チアミン : 1.000 〃 **メタルミックス水溶液 Na2 EDTA : 1.00 g/リットル−溶液 FeCl2 ・6H2 O : 0.05 〃 H3 BO3 : 1.00 〃 MnCl2 ・4H2 O : 0.15 〃 ZnCl2 : 0.01 〃 CoCl・6H2 O : 0.005 〃* Vitamin mix aqueous solution Biotin: 0.003 g / liter-solution Thiamine: 1.000〃 ** Metal mix aqueous solution Na 2 EDTA: 1.00 g / liter-solution FeCl 2 .6H 2 O: 0.050.05 H 3 BO 3: 1.00 〃 MnCl 2 · 4H 2 O: 0.15 〃 ZnCl 2: 0.01 〃 CoCl · 6H 2 O: 0.005 〃

【0019】この乾燥藻体3.2gをクロロホルム/メ
タノール=1:1(重量)混合溶媒中でヒスコトロンに
より細胞破砕後、抽出し、0.3gの油分を得た。抽出
油分を、シリカゲルプレートを用いた薄層クロマトグラ
フィー(TLC)でクロロホルム/メタノール/酢酸混
合溶媒にて展開し、極性脂質画分と中性脂質画分とに分
離した。このシリカゲルプレートより、それぞれのスポ
ットをかき取り、クロロホルム/メタノール=1:1
(重量)混合溶媒にて再抽出したところ、極性脂質含量
が60重量%、中性脂質含量が40重量%であった。
3.2 g of the dried algal cells were crushed with Hiscotron in a mixed solvent of chloroform / methanol = 1: 1 (by weight) and then extracted to obtain 0.3 g of oil. The extracted oil was developed with a mixed solvent of chloroform / methanol / acetic acid by thin layer chromatography (TLC) using a silica gel plate to separate a polar lipid fraction and a neutral lipid fraction. Each spot was scraped off from this silica gel plate, and chloroform / methanol = 1: 1.
(Weight) When reextracted with a mixed solvent, the polar lipid content was 60% by weight and the neutral lipid content was 40% by weight.

【0020】また、抽出油分200mgに硫酸/メタノー
ル混液約5mlを加えて湯浴中で1.5時間還流し、メチ
ルエステル化した後、これにヘキサンおよび水各5mlを
加えて抽出し、脂肪酸組成をガスクロマトグラフィー
(GLC)で分析したところ、総脂肪酸中のDHA含量
は42重量%であった。さらに上記極性脂質画分を同様
に処理し構成脂肪酸組成をGLC分析した結果、極性脂
質中のDHA含量は50重量%であった。
About 200 ml of the extracted oil was added with about 5 ml of a mixed solution of sulfuric acid / methanol and refluxed in a hot water bath for 1.5 hours. After methyl esterification, hexane and water (5 ml each) were added, and the mixture was extracted. Was analyzed by gas chromatography (GLC) to find that the DHA content in the total fatty acids was 42% by weight. Further, the polar lipid fraction was treated in the same manner and the constituent fatty acid composition was analyzed by GLC. As a result, the DHA content in the polar lipid was 50% by weight.

【0021】参考例2 クリプセコディニウム コーニー(ATCC3033
6)を、培養時間を定常期(124時間)とする以外は
参考例1と同じ条件で培養した。乾燥藻体の収量は50
gであった。この乾燥藻体5.0gを用いて参考例1と
同様に油分抽出、TLCおよびGLC分析した結果、油
分収量:0.7g、極性脂質含量:25重量%、中性脂
質含量:75重量%、油分の総脂肪酸中のDHA含量:
18重量%、極性脂質中のDHA含量:50重量%であ
った。
Reference Example 2 Crypsecodinium coney (ATCC3033)
6) was cultured under the same conditions as in Reference Example 1 except that the culture time was set to the stationary phase (124 hours). The yield of dried alga bodies is 50
g. As a result of oil extraction, TLC and GLC analysis using 5.0 g of the dried algal cells in the same manner as in Reference Example 1, oil yield: 0.7 g, polar lipid content: 25% by weight, neutral lipid content: 75% by weight, DHA content in total fatty acids of oil:
18% by weight, DHA content in polar lipid: 50% by weight.

【0022】参考例3 トラウストキトリウム アウレウム(ATCC3430
4)を参考例1と同様に培養した。ただし、培養中、培
養液の濁度から求めた増殖曲線において、対数増殖中期
に相当する培養開始後120時間目に、食塩20g/リ
ットルを添加して培養液の食塩濃度を2重量%上昇さ
せ、さらに24時間培養した。乾燥藻体の収量は65g
であった。この乾燥藻体6.5gを用いて参考例1と同
様に油分抽出、TLCおよびGLC分析した結果、油分
収量=0.7g、極性脂質含量:58重量%、中性脂質
含量:42重量%、油分の総脂肪酸中のDHA含量:5
0重量%、極性脂質中のDHA含量:65重量%であっ
た。
Reference Example 3 Traustochytrium aureum (ATCC3430)
4) was cultured in the same manner as in Reference Example 1. However, during the culture, in the growth curve obtained from the turbidity of the culture solution, at 120 hours after the start of the culture corresponding to the mid-logarithmic growth period, 20 g / liter of sodium chloride was added to increase the salt concentration of the culture solution by 2% by weight. And further cultured for 24 hours. 65 g of dried alga body
Met. As a result of oil extraction, TLC and GLC analysis using 6.5 g of the dried algal cells in the same manner as in Reference Example 1, oil yield = 0.7 g, polar lipid content: 58% by weight, neutral lipid content: 42% by weight, DHA content in total fatty acids of oil: 5
0% by weight, DHA content in polar lipid: 65% by weight.

【0023】実施例1 参考例1および2の方法で得た藻体を用いてシオミズツ
ボワムシを培養した。70%海水を入れた20リットル
水槽2個を20℃に調節し、通気量を0.3V/V/M
(培養液1リットル当りの毎分の空気吹込量(リット
ル))に設定した後、シオミズツボワムシ(S型および
L型の混合)を培養開始時100個体/mlになるように
セットした。ついで各水槽に、参考例1および2で得た
培養藻体(それぞれ藻体およびという)を0.1g
(湿藻体重量)/リットルとなるように投与し、経時的
に1mlあたりのシオミズツボワムシの個体数を計測し
た。その結果を図1に示す。
Example 1 A rotifer was produced using the alga bodies obtained by the methods of Reference Examples 1 and 2. Two 20-liter water tanks containing 70% seawater were adjusted to 20 ° C., and the ventilation rate was 0.3 V / V / M.
After setting the air blowing rate (liter per minute per liter of culture solution (liter)), the rotifer (mixture of S-type and L-type) was set at 100 individuals / ml at the start of the culture. Then, 0.1 g of the cultured alga bodies (referred to as alga bodies) obtained in Reference Examples 1 and 2 were added to each aquarium.
(Wet algae body weight) / liter, and the number of rotifers per ml was measured over time. The result is shown in FIG.

【0024】また、72時間培養したシオミズツボワム
シをナイロンネットで回収し、水洗して前記藻体を除去
し、シオミズツボワムシ中の脂質の脂肪酸組成を参考例
1と同様の方法で分析したところ、総脂肪酸中のDHA
含量は藻体を用いたとき28重%、藻体を用いたと
き18重量%であった。この両値の差から極性脂質の有
効性が示された。また、シオミズツボワムシの携卵率
は、藻体を飼料とした場合に50%、藻体を飼料と
した場合には45%となり、いずれも高い活力を有して
いた。
The rotifer, cultured for 72 hours, was collected with a nylon net, washed with water to remove the algal cells, and the fatty acid composition of lipids in the rotifer was analyzed in the same manner as in Reference Example 1. As a result, DHA in total fatty acids
The content was 28% by weight when using algal cells, and 18% by weight when using algal cells. The difference between these two values indicated the effectiveness of the polar lipid. The egg-bearing rate of the rotifer was 50% when the algal body was used as a feed, and 45% when the algal body was used as a feed, all of which had high vitality.

【0025】実施例2 米国産アルテミア耐久卵を水温25〜27℃の75〜1
00%海水中に入れ、卵が沈降しない程度に強力に空気
を吹込み、24時間後、孵化したアルテミアノープリウ
スと卵殻とを分離した。海水を満たした5リットル水槽
を27℃に維持し、0.3V/V/Mで空気を通気しな
がら、前記アルテミアノープリウスを100個体/mlの
初発濃度になるようにセットした。これに参考例1の方
法で得た生藻体を0.2g(湿藻体重量)/リットルと
なるように投与し、24時間培養し摂餌させた。参考例
1の方法に準拠し、アルテミア中の脂質の脂肪酸組成を
分析したところ、孵化直後すなわち藻類での培養開始前
では総脂肪酸中のDHA含量が1重量%以下であった
が、藻類で24時間培養したものでは29重量%に上昇
していた。
Example 2 US-made durable Artemia eggs were cultivated at 75-1 ° C at a water temperature of 25-27 ° C.
After putting in 00% seawater and blowing air so strongly that the eggs did not settle, 24 hours later, the hatched Artemia nauplius and the eggshell were separated. While maintaining a 5-liter water tank filled with seawater at 27 ° C. and ventilating air at 0.3 V / V / M, the above-mentioned Artemia nauplius was set to an initial concentration of 100 individuals / ml. The fresh algal cells obtained by the method of Reference Example 1 were administered thereto at 0.2 g (wet algal cell weight) / liter, cultured for 24 hours and fed. When the fatty acid composition of the lipid in Artemia was analyzed in accordance with the method of Reference Example 1, the DHA content in the total fatty acids was 1% by weight or less immediately after hatching, that is, before the start of cultivation in algae. After culturing for an hour, the content increased to 29% by weight.

【0026】実施例3 50%海水を満たした5リットル水槽を25℃に維持
し、0.3V/V/Mで空気を通気しながら、汽水ミジ
ンコを50個体/mlの初発濃度になるようにセットし
た。これに参考例3の方法で得た乾燥藻体を0.1g
(乾燥藻体重量)/リットルとなるように投入し、60
時間培養し摂餌させた。参考例1の方法に準拠し、ミジ
ンコ中の脂質の脂肪酸組成を分析したところ、当初の総
脂肪酸中のDHA含量は0.8重量%であり、ほとんど
含有されていなかったが、藻類で培養したものでは25
重量%まで強化されていた。
Example 3 A 5-liter aquarium filled with 50% seawater was maintained at 25 ° C., and air was supplied at a flow rate of 0.3 V / V / M so that the initial concentration of brackish water daphnia was 50 individuals / ml. I set it. 0.1 g of the dried algal cells obtained by the method of Reference Example 3
(Dry algae body weight) / liter
They were cultured for a period of time and fed. When the fatty acid composition of the lipids in the daphnia was analyzed in accordance with the method of Reference Example 1, the initial DHA content in the total fatty acids was 0.8% by weight, and the DHA content was very low. 25
It was fortified to weight percent.

【0027】実施例4 クリプセコディニウム コーニー(ATCC3033
6)を参考例1と同条件で培養し、湿藻体を得た。この
湿藻体の一部を−20℃で凍結し、また常法により凍結
乾燥した。実施例1に記載のシオミズツボワムシの培養
方法を用い、培養開始時のシオミズツボワムシの濃度を
100個体/mlに調整し、前記の湿藻体、凍結藻体また
は凍結乾燥藻体を各0.1g(乾燥藻体重量換算)/リ
ットルとなるように投入し、それぞれ同様に培養した。
その結果、シオミズツボワムシの増殖速度はいずれもほ
ぼ同程度であり、72時間培養後のシオミズツボワムシ
の脂質の総脂肪酸中のDHA含量は25〜28重量%
(湿藻体のとき28重量%、凍結藻体のとき26重量
%、凍結乾燥藻体のとき25重量%)であり、携卵率も
45〜50%であった。
Example 4 Crypsecodinium coney (ATCC 3033)
6) was cultured under the same conditions as in Reference Example 1 to obtain a wet algal body. A part of this wet algal body was frozen at -20 ° C and freeze-dried by a conventional method. By using the method of cultivating the rotifer of the rotifer described in Example 1, the concentration of the rotifer at the start of the cultivation was adjusted to 100 individuals / ml, and the wet alga body, the frozen alga body, or the lyophilized alga body was obtained. 0.1 g (in terms of dry alga body weight) / liter was added, and the cells were cultured in the same manner.
As a result, the growth rates of the rotifer were almost the same, and the DHA content in the total fatty acids of the lipid of the rotifer was 25 to 28% by weight after culturing for 72 hours.
(28% by weight for wet algal bodies, 26% by weight for frozen algal bodies, 25% by weight for freeze-dried algal bodies), and the egg carrying rate was 45 to 50%.

【0028】比較例1 実施例1のシオミズツボワムシの培養方法において、本
発明に係わる藻体0.1g/リットルを油脂酵母(協和
発酵(株)製、製品名「油脂酵母」:魚油を資化させた
酵母)0.5g/リットルに置き換え、他の条件を同一
にして培養した。72時間培養後のシオミズツボワムシ
の脂質における総脂肪酸中のDHA含量は2.5重量%
にすぎず、携卵率は35%であった。
Comparative Example 1 In the method for cultivating the rotifer, Rotifer, 0.1 g / l of the alga body according to the present invention was prepared by using a fat oil yeast (manufactured by Kyowa Hakko Co., Ltd., product name "oil fat yeast": fish oil). (Assimilated yeast) was replaced with 0.5 g / liter and cultured under the same other conditions. DHA content in total fatty acids in the lipid of Rotifer, Rotifer was 72% by weight after culturing for 72 hours.
The egg-carrying rate was only 35%.

【0029】比較例2 実施例1のシオミズツボワムシの培養方法において、本
発明に係わる藻体0.1g/リットルを淡水産クロレラ
(日本クロレラ(株)製、製品名「フレッシュグリーン
600」:従属栄養で培養した淡水産クロレラの生藻
体)0.3g/リットルに置き換え、他の条件を同一に
して培養した。シオミズツボワムシの増殖速度および携
卵率は実施例1の結果とほぼ同程度であったが、シオミ
ズツボワムシの脂質における総脂肪酸中のDHA含量
は、120時間培養後でも0.5重量%以下であった。
Comparative Example 2 In the method for cultivating the rotifer, Rotifer, 0.1 g / l according to the present invention, freshwater chlorella (manufactured by Nippon Chlorella, product name "Fresh Green 600"): The fresh algae of freshwater chlorella cultured by nutrient) were replaced with 0.3 g / liter, and cultured under the same other conditions. The growth rate and egg carrying rate of the rotifer were almost the same as the results of Example 1, but the DHA content in the total fatty acids in the lipid of the rotifer was 0.5 weight even after culturing for 120 hours. % Or less.

【0030】比較例3 実施例1のシオミズツボワムシの培養方法において、本
発明に係わる藻体0.1g/リットルを粉末精製魚油
(キユーピー(株)製、製品名「シーオイルパウダ
ー」:DHA含量15重量%の魚油を卵白ペプチドに吸
着させたもの)0.5g/リットルに置き換え、他の条
件を同一にして培養した。72時間培養後のシオミズツ
ボワムシの脂質における総脂肪酸中のDHA含量は5重
量%にすぎず、携卵率は30%であった。
Comparative Example 3 In the method for cultivating the rotifer, Rotifer, 0.1 g / l of the algae according to the present invention, powdered fish oil (product name "Sea Oil Powder", manufactured by KUP Co., Ltd .: DHA) A 15% by weight fish oil adsorbed on an egg white peptide) was replaced with 0.5 g / liter and cultured under the same other conditions. The DHA content in the total fatty acids in the lipid of the rotifer, Rotifer rotifer after 72 hours of cultivation was only 5% by weight, and the egg-carrying rate was 30%.

【0031】[0031]

【発明の効果】本発明によれば、水産種苗生産における
稚仔魚の生物餌料である動物性プランクトンを養成する
ための有用な飼料が提供される。本発明の飼料は、従属
栄養で培養できる藻類を用いるため安価に製造でき、動
物性プランクトンにDHAを効率的に栄養強化でき、そ
の含有量は動物性プランクトンの脂質の総脂肪酸のうち
約20〜30重量%に達する。また携卵率が高く、活力
の高い動物性プランクトンを安定的に養成することがで
きる。
According to the present invention, there is provided a useful feed for nurturing zooplankton, which is a biological feed for juvenile larvae in the production of marine seedlings. The feed of the present invention can be produced at low cost by using algae that can be cultured by heterotrophic nutrition, can efficiently fortify DHA to zooplankton, and its content is about 20 to 20% of the total fatty acids of lipids of zooplankton. Reaches 30% by weight. In addition, the zooplankton, which has a high egg carrying rate and high vitality, can be stably cultivated.

【図面の簡単な説明】[Brief description of the drawings]

【図1】 単位培養液あたりのシオミズツボワムシの個
体数と培養時間との関係を表す図である。藻体は参考
例1の方法で、藻体は参考例2の方法でそれぞれ得た
藻体を意味する。
FIG. 1 is a diagram showing the relationship between the number of rotifers per unit culture and the cultivation time. Algae refers to the alga body obtained by the method of Reference Example 1, and alga body refers to the alga body obtained by the method of Reference Example 2.

Claims (6)

(57)【特許請求の範囲】(57) [Claims] 【請求項1】 従属栄養で生育し、ドコサヘキサエン酸
含有極性脂質を含み、かつ総脂肪酸中のドコサヘキサエ
ン酸含量が10重量%以上である藻類を原料としてなる
動物性プランクトン培養用飼料。
1. A feed for cultivating zooplankton, which is grown under heterotrophic conditions, contains docosahexaenoic acid-containing polar lipids, and has a docosahexaenoic acid content in total fatty acids of at least 10% by weight as an ingredient.
【請求項2】 藻類の脂質中、前記極性脂質の含有量が
30重量%以上である請求項1に記載の飼料。
2. The feed according to claim 1, wherein the content of the polar lipid in the algal lipid is 30% by weight or more.
【請求項3】 藻類の脂質中、前記極性脂質の構成脂肪
酸のうちドコサヘキサエン酸含有量が30重量%以上で
ある請求項1または2に記載の飼料。
3. The feed according to claim 1, wherein the content of docosahexaenoic acid among the fatty acids constituting the polar lipid in the algal lipid is 30% by weight or more.
【請求項4】 藻類がクリプセコディニウム(Crypthec
odinium )属に属する単細胞藻類である請求項1〜3の
いずれかに記載の飼料。
4. The method according to claim 1, wherein the algae is Crypthecdinium.
The feed according to any one of claims 1 to 3, which is a unicellular algae belonging to the genus odinium).
【請求項5】 藻類が、その培養における指数増殖中期
以降に、生育に好適な食塩濃度より0.1〜10重量%
高い食塩濃度で培養されたものである請求項1〜4のい
ずれかに記載の飼料。
5. The method according to claim 5, wherein the algae is 0.1 to 10% by weight of a salt concentration suitable for growth after the middle of exponential growth in the culture.
The feed according to any one of claims 1 to 4, wherein the feed is cultured at a high salt concentration.
【請求項6】 動物性プランクトンがシオミズツボワム
シ、アルテミア、ミジンコである請求項1〜5のいずれ
かに記載の飼料。
6. The feed according to claim 1, wherein the zooplankton is a rotifer, Artemia or Daphnia.
JP24885193A 1993-09-09 1993-09-09 Animal Plankton Culture Feed Expired - Fee Related JP3302123B2 (en)

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JPH0775507A JPH0775507A (en) 1995-03-20
JP3302123B2 true JP3302123B2 (en) 2002-07-15

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ID=17184373

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE305048T1 (en) * 1997-08-01 2005-10-15 Martek Biosciences Corp NUTRIENT COMPOSITIONS CONTAINING DHA AND METHOD FOR THE PRODUCTION THEREOF
US6399118B1 (en) * 2001-06-29 2002-06-04 Fish Biotech Ltd. Process for storing enriched nematodes
JP5415299B2 (en) * 2010-01-14 2014-02-12 学校法人近畿大学 Biological feed for flounder breeding and flounder breeding method
KR101934552B1 (en) * 2018-05-09 2019-03-25 어업회사법인 가비 주식회사 Method of Improving Survival Rate for Hatching Fry of Coldsea Fish
JP2023166636A (en) * 2020-10-06 2023-11-22 Dic株式会社 Feed composition for zooplankton, production method of zooplankton, zooplankton, production method of fishery living thing, and additive to rearing water for fishery living thing

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