JP6822258B2 - Rotifer feed - Google Patents
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- JP6822258B2 JP6822258B2 JP2017057317A JP2017057317A JP6822258B2 JP 6822258 B2 JP6822258 B2 JP 6822258B2 JP 2017057317 A JP2017057317 A JP 2017057317A JP 2017057317 A JP2017057317 A JP 2017057317A JP 6822258 B2 JP6822258 B2 JP 6822258B2
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- 241000700141 Rotifera Species 0.000 title claims description 103
- 230000002378 acidificating effect Effects 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 3
- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 description 16
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 14
- 238000004519 manufacturing process Methods 0.000 description 13
- 230000007704 transition Effects 0.000 description 9
- 235000020661 alpha-linolenic acid Nutrition 0.000 description 8
- 229960004488 linolenic acid Drugs 0.000 description 8
- 238000000034 method Methods 0.000 description 7
- 229910052757 nitrogen Inorganic materials 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 235000005911 diet Nutrition 0.000 description 6
- 230000037213 diet Effects 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 239000012141 concentrate Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000003860 storage Methods 0.000 description 5
- 238000011156 evaluation Methods 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 229930003779 Vitamin B12 Natural products 0.000 description 2
- 238000005273 aeration Methods 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- AGVAZMGAQJOSFJ-WZHZPDAFSA-M cobalt(2+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+2].N#[C-].[N-]([C@@H]1[C@H](CC(N)=O)[C@@]2(C)CCC(=O)NC[C@@H](C)OP(O)(=O)O[C@H]3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)\C2=C(C)/C([C@H](C\2(C)C)CCC(N)=O)=N/C/2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O AGVAZMGAQJOSFJ-WZHZPDAFSA-M 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 150000002739 metals Chemical class 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 229960003495 thiamine Drugs 0.000 description 2
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 235000019163 vitamin B12 Nutrition 0.000 description 2
- 239000011715 vitamin B12 Substances 0.000 description 2
- 241000700104 Brachionus plicatilis Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241001415279 Pseudococcidae Species 0.000 description 1
- 241001140502 Pseudococcomyxa Species 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 229930003451 Vitamin B1 Natural products 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- LFYJSSARVMHQJB-QIXNEVBVSA-N bakuchiol Chemical compound CC(C)=CCC[C@@](C)(C=C)\C=C\C1=CC=C(O)C=C1 LFYJSSARVMHQJB-QIXNEVBVSA-N 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 235000020774 essential nutrients Nutrition 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229960002901 sodium glycerophosphate Drugs 0.000 description 1
- REULQIKBNNDNDX-UHFFFAOYSA-M sodium;2,3-dihydroxypropyl hydrogen phosphate Chemical compound [Na+].OCC(O)COP(O)([O-])=O REULQIKBNNDNDX-UHFFFAOYSA-M 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 229910021654 trace metal Inorganic materials 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000019158 vitamin B6 Nutrition 0.000 description 1
- 239000011726 vitamin B6 Substances 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
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- Feed For Specific Animals (AREA)
- Fodder In General (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
本開示はワムシ用餌料及びその製造方法に関する。 The present disclosure relates to feed for rotifers and methods for producing the same.
シオミズツボワムシ(以下、ワムシとする)は、動物プランクトンの一種である。養殖産業において種苗生産対象稚仔を飼育するとき、ワムシは初期餌料として使用される。養殖産業においてワムシは大量に使用されるため、ワムシを大量に培養する必要がある。ワムシの培養において給餌されるワムシ用餌料として、産業用微細藻類濃縮液が使用される。産業用微細藻類をワムシの培養に用いることは特許文献１に開示されている。 Brachionus plicatilis (hereinafter referred to as rotifer) is a type of zooplankton. Rotifers are used as the initial feed when raising seedling-producing juveniles in the aquaculture industry. Since rotifers are used in large quantities in the aquaculture industry, it is necessary to cultivate rotifers in large quantities. An industrial microalgae concentrate is used as a feed for rotifers to be fed in rotifer culture. The use of industrial microalgae for rotifer culture is disclosed in Patent Document 1.
現在、産業用微細藻類濃縮液は、製造後の保存可能な期間が限られるため、さらに長期的な保存が可能になること望まれる。
本開示は、保存可能な期間が長いワムシ用餌料及びその製造方法を提供する。
At present, the industrial microalgae concentrate has a limited shelf life after production, so it is desired that it can be stored for a longer period of time.
The present disclosure provides a feed for rotifers having a long shelf life and a method for producing the same.
本開示の一態様は、微細藻類シュードココミクサ属を含むワムシ用餌料である。本開示の一態様であるワムシ用餌料は、ワムシを増殖させる効果が高い。また、本開示の一態様であるワムシ用餌料は、保存可能な期間が長い。 One aspect of the present disclosure is a feed for rotifers containing the microalgae Pseudococomixa. The rotifer feed, which is one aspect of the present disclosure, has a high effect of growing rotifers. In addition, the rotifer feed, which is one aspect of the present disclosure, has a long shelf life.
本開示の別の態様は、微細藻類シュードココミクサ属を培地において培養し、前記培地に含まれる水の少なくとも一部を除去するワムシ用餌料の製造方法である。本開示の別の態様であるワムシ用餌料の製造方法によれば、ワムシを増殖させる効果が高く、保存可能な期間が長いワムシ用餌料を製造することができる。 Another aspect of the present disclosure is a method for producing a feed for rotifers in which microalgae Pseudococcidae are cultured in a medium and at least a part of water contained in the medium is removed. According to the method for producing a rotifer feed according to another aspect of the present disclosure, it is possible to produce a rotifer feed having a high effect of growing rotifers and having a long shelf life.
本開示の実施形態を説明する。
１．ワムシ用餌料
ワムシ用餌料は、微細藻類シュードココミクサ(Pseudococcomyxa)属を含む。微細藻類シュードココミクサ属として、適宜選択した株を用いることができる。好ましい株として、例えば、シュードココミクサ ｓｐ.ＫＪ株等が挙げられる。
An embodiment of the present disclosure will be described.
1. 1. Rotifer diets Rotifer diets include the genus Pseudococcomyxa, a microalgae. As the genus Pseudococomixa, a strain appropriately selected can be used. Preferred strains include, for example, Pseudococomixa sp.KJ strain and the like.
シュードココミクサ ｓｐ.ＫＪ株は、２０１３年６月４日付で独立行政法人製品評価技術基盤機構 特許生物寄託センター（ＮＩＴＥ−ＩＰＯＤ）（千葉県木更津市かずさ鎌足２−５−８ １２０号室）に受託番号ＦＥＲＭ Ｐ−２２２５４として寄託されている。 Pseudo Cocomixa sp.KJ strain was transferred to the National Institute of Technology and Evaluation Patent Organism Depositary (NITE-IPOD) (Room 2-5-8 Kazusakamatari, Kisarazu City, Chiba Prefecture) on June 4, 2013. It has been deposited under the accession number FERM P-22254.
ワムシ用餌料の剤型は特に限定されず、例えば、液状、粉末状等が挙げられる。ワムシ用餌料は酸性であることが好ましい。酸性である場合、ワムシ用餌料におけるコンタミ微生物の増殖を抑制できる。ワムシ用餌料のｐＨは４以下であることが好ましく、３以上４以下であることが特に好ましい。ｐＨがこの範囲内である場合、ワムシ用餌料におけるコンタミ微生物の増殖を一層抑制できる。 The dosage form of the rotifer feed is not particularly limited, and examples thereof include liquid and powder. The rotifer feed is preferably acidic. When acidic, the growth of contaminated microorganisms in the rotifer diet can be suppressed. The pH of the rotifer feed is preferably 4 or less, and particularly preferably 3 or more and 4 or less. When the pH is within this range, the growth of contaminated microorganisms in the rotifer diet can be further suppressed.
ワムシ用餌料の剤型が液状である場合、ワムシ用餌料における微細藻類シュードココミクサ属の濃度は、１００〜１５０（乾燥重量）ｇ／Ｌの範囲が好ましい。
ワムシ用餌料は中性であってもよい。微細藻類シュードココミクサ属を培養する培地が酸性である場合、例えば、培養終了後の培地に塩基性物質を添加し、中和することで、ワムシ用餌料を中性にすることができる。塩基性物質として、例えば、水酸化ナトリウム等が挙げられる。
When the dosage form of the rotifer feed is liquid, the concentration of the microalga Pseudococomixa in the rotifer feed is preferably in the range of 100 to 150 (dry weight) g / L.
The rotifer diet may be neutral. When the medium for culturing the microalgae Pseudococcylus is acidic, for example, by adding a basic substance to the medium after the culture is completed and neutralizing it, the rotifer feed can be neutralized. Examples of the basic substance include sodium hydroxide and the like.
２．ワムシ用餌料の製造方法
ワムシ用餌料の製造方法では、微細藻類シュードココミクサ属を培地において培養し、前記培地に含まれる水の少なくとも一部を除去する。
2. 2. Method for Producing Rotifer Feed In the method for producing rotifer feed, the genus Pseudococomixa, a microalgae, is cultured in a medium and at least a part of water contained in the medium is removed.
微細藻類シュードココミクサ属として、適宜選択した株を用いることができる。好ましい株として、例えば、シュードココミクサ ｓｐ.ＫＪ株等が挙げられる。
培地として、微細藻類の培養に通常使用されているものを使用することができる。培地として、例えば、各種栄養塩、微量金属塩、ビタミン等を含む公知の淡水産微細藻類用の培地、海産微細藻類用の培地等を使用することができる。培地として、例えば、ＡＦ６培地が挙げられる。ＡＦ６培地の組成は以下のとおりである。以下の数値は１００ｍｌの培地に含まれる質量である。
As the genus Pseudococomixa, a strain appropriately selected can be used. Preferred strains include, for example, Pseudococomixa sp.KJ strain and the like.
As the medium, a medium usually used for culturing microalgae can be used. As the medium, for example, a known medium for freshwater microalgae containing various nutrient salts, trace metal salts, vitamins and the like, a medium for marine microalgae and the like can be used. Examples of the medium include AF6 medium. The composition of AF6 medium is as follows. The following numbers are the mass contained in 100 ml of medium.
NaNO3：14mg
NH4NO3：2.2mg
MgSO4・7H2O：3mg
KH2PO4：1mg
K2HPO4：0.5mg
CaCl2・2H2O：1mg
CaCO3：1mg
Fe-citrate：0.2mg
Citric acid：0.2mg
Biotin：0.2μg
Thiamine HCl：1μg
Vitamin B6：0.1μg
Vitamin B12：0.1μg
Trace metals：0.5mL
Distilled water：99.5mL
栄養塩として、例えば、ＮａＮＯ３、ＫＮＯ３、ＮＨ４Ｃｌ、尿素等の窒素源；Ｋ２ＨＰＯ４、ＫＨ２ＰＯ４、グリセロリン酸ナトリウム等のリン源が挙げられる。また、微量金属として、例えば、鉄、マグネシウム、マンガン、カルシウム、亜鉛等が挙げられる。ビタミンとして、例えば、ビタミンＢ１、ビタミンＢ１２等が挙げられる。
NaNO 3 : 14 mg
NH 4 NO 3 : 2.2 mg
MgSO 4 · 7H 2 O: 3mg
KH 2 PO 4 : 1 mg
K 2 HPO 4 : 0.5 mg
CaCl 2 · 2H 2 O: 1mg
CaCO 3 : 1 mg
Fe-citrate: 0.2mg
Citric acid: 0.2mg
Biotin: 0.2 μg
Thiamine HCl: 1 μg
Vitamin B6: 0.1 μg
Vitamin B12: 0.1 μg
Trace metals: 0.5mL
Distilled water: 99.5mL
Examples of nutrient salts include nitrogen sources such as NaNO 3 , KNO 3 , NH 4 Cl, and urea; and phosphorus sources such as K 2 HPO 4 , KH 2 PO 4 , and sodium glycerophosphate. Examples of trace metals include iron, magnesium, manganese, calcium, zinc and the like. Examples of vitamins include vitamin B1 and vitamin B12.
微細藻類シュードココミクサ属の培養方法は、例えば、通気条件で二酸化炭素の供給とともに攪拌を行う方法である。その際、例えば、明暗サイクルをつけた光照射、又は、連続光照射して培養することができる。明暗サイクルとして、例えば、蛍光灯での１２時間の光照射と、１２時間の暗条件とを繰り返す明暗サイクル等が挙げられる。 The method for culturing the microalgae Pseudococomixa is, for example, a method of supplying carbon dioxide and stirring under aeration conditions. At that time, for example, light irradiation with a light-dark cycle or continuous light irradiation can be used for culturing. Examples of the light-dark cycle include a light-dark cycle in which a 12-hour light irradiation with a fluorescent lamp and a 12-hour dark condition are repeated.
培地は酸性であることが好ましい。培地が酸性である場合、培地におけるコンタミ微生物の増殖を抑制できる。培地のｐＨが４以下であることが好ましく、３以上４以下であることが特に好ましい。ｐＨがこの範囲内である場合、培地におけるコンタミ微生物の増殖を一層抑制できる。培養温度は、２０〜３０℃であることが好ましい。 The medium is preferably acidic. When the medium is acidic, the growth of contaminated microorganisms in the medium can be suppressed. The pH of the medium is preferably 4 or less, and particularly preferably 3 or more and 4 or less. When the pH is within this range, the growth of contaminated microorganisms in the medium can be further suppressed. The culture temperature is preferably 20 to 30 ° C.
培地は窒素十分の状態であることが好ましい。培地が窒素十分の状態である場合、微細藻類シュードココミクサ属に含まれるα―リノレン酸の量を増すことができる。α―リノレン酸は、種苗生産対象稚仔の必須栄養素の１つであると考えられている。 The medium is preferably in a state of sufficient nitrogen. When the medium is nitrogen-sufficient, the amount of α-linolenic acid contained in the microalgae Pseudococomixa can be increased. α-linolenic acid is considered to be one of the essential nutrients for seedling production target infants.
窒素十分の状態とは、例えば、Ｎ／ＤＷが２質量％以上の状態である。窒素欠乏の状態とは、例えば、Ｎ／ＤＷが２質量％未満の状態である。Ｎ／ＤＷは、藻体の乾燥重量に占める窒素重量の比率である。 The state of sufficient nitrogen is, for example, a state in which N / DW is 2% by mass or more. The nitrogen-deficient state is, for example, a state in which N / DW is less than 2% by mass. N / DW is the ratio of nitrogen weight to dry weight of algae.
微細藻類シュードココミクサ属に含まれるα―リノレン酸の量が増すと、本開示のワムシ用餌料を給餌したワムシに含まれるα―リノレン酸の量が増加する。その結果、ワムシを給餌した種苗生産対象稚仔の生存率を高めることができる。 As the amount of α-linolenic acid contained in the microalgae Pseudococomixa increases, the amount of α-linolenic acid contained in the rotifers fed the rotifer diet of the present disclosure increases. As a result, the survival rate of the seedling production target larvae fed with rotifer can be increased.
培地に含まれる水の少なくとも一部を除去する工程は、例えば、遠心分離、膜分離等の方法で行うことができる。培地に含まれる水の少なくとも一部を除去した結果、微細藻類シュードココミクサ属は濃縮される。 The step of removing at least a part of the water contained in the medium can be performed by, for example, a method such as centrifugation or membrane separation. As a result of removing at least a part of the water contained in the medium, the microalgae Pseudococomixa is concentrated.
３．ワムシ用餌料及びその製造方法が奏する効果
（１Ａ）本開示のワムシ用餌料は、ワムシを増殖させる効果が高い。
（１Ｂ）本開示のワムシ用餌料は、保存可能な期間が長い。
3. 3. Effects of rotifer feed and its production method (1A) The rotifer feed of the present disclosure has a high effect of growing rotifers.
(1B) The rotifer feed of the present disclosure has a long shelf life.
（１Ｃ）本開示のワムシ用餌料の製造方法によれば、微細藻類シュードココミクサ属を含むワムシ用餌料を製造することができる。製造したワムシ用餌料は、ワムシを増殖させる効果が高く、保存可能な期間が長い。 (1C) According to the method for producing a rotifer feed of the present disclosure, a rotifer feed containing the microalgae Pseudococomixa can be produced. The produced rotifer feed has a high effect of growing rotifers and has a long shelf life.
４．実施例
（４−１）ワムシ用餌料の製造
以下の条件で、シュードココミクサ ｓｐ.ＫＪ株を培養した。培養の具体的な手順は以下のとおりとした。
4. Example (4-1) Production of feed for rotifers Pseudococomixa sp.KJ strain was cultured under the following conditions. The specific procedure for culturing was as follows.
（i）ＡＦ６培地を調製する。培地は窒素十分条件とする。培地のｐＨは３．５である。
（ii）１２１℃、２０ｍｉｎの条件で培地を滅菌し、シュードココミクサ ｓｐ.ＫＪ株を植藻する。
(I) Prepare AF6 medium. The medium should be nitrogen sufficient. The pH of the medium is 3.5.
(Ii) Sterilize the medium under the conditions of 121 ° C. and 20 min, and inoculate the pseudococomixa sp.KJ strain.
（iii）２％Ｃ０２を通気する
（iv）光を照射しながら室温にて培養する。光の強さは約１００μmol/m2/sとする。室温は２５℃である。
(Iii) a 2% C0 2 was irradiated with aeration to (iv) light while culturing at room temperature. The light intensity is about 100 μmol / m 2 / s. Room temperature is 25 ° C.
培養終了後、培地に含まれる水の一部を遠心分離により除去し、ワムシ用餌料Ｓ１を完成した。ワムシ用餌料Ｓ１におけるシュードココミクサ ｓｐ.ＫＪ株の濃度は、１４０（乾燥重量）ｇ／Ｌである。 After completion of the culture, a part of the water contained in the medium was removed by centrifugation to complete the rotifer feed S1. The concentration of the pseudococomixa sp.KJ strain in the rotifer feed S1 is 140 (dry weight) g / L.
また、基本的にはワムシ用餌料Ｓ１と同様にしてワムシ用餌料Ｓ２を製造した。ただし、ワムシ用餌料Ｓ２の場合、窒素欠乏条件の培地でシュードココミクサ ｓｐ.ＫＪ株を培養した。 Further, basically, the rotifer feed S2 was produced in the same manner as the rotifer feed S1. However, in the case of the rotifer feed S2, the pseudococomixa sp.KJ strain was cultured in a medium under nitrogen-deficient conditions.
また、基本的にはワムシ用餌料Ｓ１と同様にしてワムシ用餌料Ｓ３を製造した。ただし、ワムシ用餌料Ｓ３の場合、培養終了後に水酸化ナトリウムを適量加えて、中性にした。
（４−２）ワムシ用餌料の分析
ガスクロマトグラフを用いて、ワムシ用餌料Ｓ１、Ｓ２におけるα―リノレン酸の量を測定した。その結果を図１に示す。図１に示す量は、１００ｇのシュードココミクサ ｓｐ.ＫＪ株に含まれるα―リノレン酸の質量（単位はｇ）である。ワムシ用餌料Ｓ１では、ワムシ用餌料Ｓ２に比べて、α―リノレン酸の量が多かった。
Further, basically, the rotifer feed S3 was produced in the same manner as the rotifer feed S1. However, in the case of the rotifer feed S3, an appropriate amount of sodium hydroxide was added after the completion of the culture to make it neutral.
(4-2) Analysis of rotifer feed The amount of α-linolenic acid in rotifer feeds S1 and S2 was measured using a gas chromatograph. The result is shown in FIG. The amount shown in FIG. 1 is the mass (unit: g) of α-linolenic acid contained in 100 g of Pseudococomixa sp.KJ strain. The amount of α-linolenic acid in the rotifer feed S1 was larger than that in the rotifer feed S2.
（４−３）ワムシ用餌料の使用と評価（その１）
ワムシ用餌料Ｓ１をワムシに１日１回給餌した。そのときのワムシ密度の推移を図２に示す。比較例として、産業用微細藻類濃縮液をワムシに１日１回給餌した。比較例における給餌量は、ワムシ用餌料Ｓ１の給餌量と同じである。比較例を給餌したときのワムシ密度の推移を図２に示す。
(4-3) Use and evaluation of rotifer feed (Part 1)
The rotifer feed S1 was fed to the rotifer once a day. The transition of the rotifer density at that time is shown in FIG. As a comparative example, an industrial microalgae concentrate was fed to rotifers once a day. The feed amount in the comparative example is the same as the feed amount of the rotifer feed S1. FIG. 2 shows the transition of the rotifer density when the comparative example was fed.
ワムシ用餌料Ｓ１、及び比較例のいずれを給餌した場合でも、ワムシ密度は増加した。培養日数が８日を越えると、ワムシ用餌料Ｓ１を給餌した場合の方が、比較例を給餌した場合よりも、ワムシの増殖が一層顕著であった。 The rotifer density increased when either the rotifer feed S1 or the comparative example was fed. When the number of days of culture exceeded 8 days, the growth of rotifers was more remarkable in the case of feeding the rotifer feed S1 than in the case of feeding the comparative example.
（４−４）ワムシ用餌料の使用と評価（その２）
ワムシ用餌料Ｓ１、Ｓ３を、それぞれ、製造から１ヶ月保存した。１ヶ月保存後のワムシ用餌料Ｓ１をワムシに１日１回給餌した。そのときのワムシ密度の推移を図３に示す。
(4-4) Use and evaluation of rotifer feed (Part 2)
The rotifer feeds S1 and S3 were stored for one month from production, respectively. The rotifer feed S1 after storage for one month was fed to the rotifer once a day. The transition of the rotifer density at that time is shown in FIG.
また、１ヶ月保存後のワムシ用餌料Ｓ３をワムシに１日１回給餌した。そのときのワムシ密度の推移を図３に示す。ワムシ用餌料Ｓ１、Ｓ３のいずれを給餌した場合でも、ワムシ密度は増加した。 In addition, the rotifer feed S3, which had been stored for one month, was fed to the rotifer once a day. The transition of the rotifer density at that time is shown in FIG. The rotifer density increased regardless of whether the rotifer feeds S1 and S3 were fed.
保存可能な期間を過ぎた比較例の微細藻類濃縮液をワムシに給餌した場合、ワムシは短期間で全滅した。
（４−５）ワムシ用餌料の使用と評価（その３）
ワムシ用餌料Ｓ１、Ｓ３を、それぞれ、製造から１ヶ月半保存した。１ヶ月半保存後のワムシ用餌料Ｓ１をワムシに１日１回給餌した。そのときのワムシ密度の推移を図４に示す。
When the rotifers were fed with the microalgae concentrate of the comparative example that had passed the storage period, the rotifers were annihilated in a short period of time.
(4-5) Use and evaluation of rotifer feed (Part 3)
The rotifer feeds S1 and S3 were stored for one and a half months from production, respectively. The rotifer feed S1 after storage for one and a half months was fed to the rotifer once a day. The transition of the rotifer density at that time is shown in FIG.
また、１ヶ月半保存後のワムシ用餌料Ｓ３をワムシに１日１回給餌した。そのときのワムシ密度の推移を図４に示す。ワムシ用餌料Ｓ１、Ｓ３のいずれを給餌した場合でも、ワムシ密度は増加した。 In addition, the rotifer feed S3, which had been stored for one and a half months, was fed to the rotifer once a day. The transition of the rotifer density at that time is shown in FIG. The rotifer density increased regardless of whether the rotifer feeds S1 and S3 were fed.
５．他の実施形態
以上、本開示の実施形態について説明したが、本開示は上述の実施形態に限定されることなく、種々変形して実施することができる。
5. Other Embodiments Although the embodiments of the present disclosure have been described above, the present disclosure is not limited to the above-described embodiments, and can be implemented in various modifications.
（１）上記実施形態における１つの構成要素が有する複数の機能を、複数の構成要素によって実現したり、１つの構成要素が有する１つの機能を、複数の構成要素によって実現したりしてもよい。また、複数の構成要素が有する複数の機能を、１つの構成要素によって実現したり、複数の構成要素によって実現される１つの機能を、１つの構成要素によって実現したりしてもよい。また、上記実施形態の構成の一部を省略してもよい。また、上記実施形態の構成の少なくとも一部を、他の上記実施形態の構成に対して付加又は置換してもよい。なお、特許請求の範囲に記載した文言から特定される技術思想に含まれるあらゆる態様が本開示の実施形態である。 (1) A plurality of functions possessed by one component in the above embodiment may be realized by a plurality of components, or one function possessed by one component may be realized by a plurality of components. .. Further, a plurality of functions possessed by the plurality of components may be realized by one component, or one function realized by the plurality of components may be realized by one component. Further, a part of the configuration of the above embodiment may be omitted. In addition, at least a part of the configuration of the above embodiment may be added or replaced with the configuration of the other above embodiment. It should be noted that all aspects included in the technical idea specified from the wording described in the claims are embodiments of the present disclosure.
（２）上述したワムシ用餌料の他、当該ワムシ用餌料を構成要素とする製品、ワムシの増殖方法、種苗生産対象稚仔の飼育方法等、種々の形態で本開示を実現することもできる。 (2) In addition to the above-mentioned rotifer feed, the present disclosure can be realized in various forms such as a product containing the rotifer feed as a component, a method for breeding rotifer, and a method for raising larvae to be produced as seedlings.
Claims (3)
酸性であり、液状であるワムシ用餌料。 The rotifer feed according to claim 1.
A rotifer feed that is acidic and liquid.
ｐＨが４以下であるワムシ用餌料。 The rotifer feed according to claim 2.
A feed for rotifers with a pH of 4 or less.
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