JP2014226123A - Algae culture method - Google Patents
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- JP2014226123A JP2014226123A JP2013110865A JP2013110865A JP2014226123A JP 2014226123 A JP2014226123 A JP 2014226123A JP 2013110865 A JP2013110865 A JP 2013110865A JP 2013110865 A JP2013110865 A JP 2013110865A JP 2014226123 A JP2014226123 A JP 2014226123A
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- 241000195493 Cryptophyta Species 0.000 title claims abstract description 56
- 238000012136 culture method Methods 0.000 title claims abstract description 25
- 229910001919 chlorite Inorganic materials 0.000 claims abstract description 36
- 229910052619 chlorite group Inorganic materials 0.000 claims abstract description 36
- QBWCMBCROVPCKQ-UHFFFAOYSA-M chlorite Chemical compound [O-]Cl=O QBWCMBCROVPCKQ-UHFFFAOYSA-M 0.000 claims abstract description 24
- 229940005993 chlorite ion Drugs 0.000 claims abstract description 24
- QBWCMBCROVPCKQ-UHFFFAOYSA-N chlorous acid Chemical compound OCl=O QBWCMBCROVPCKQ-UHFFFAOYSA-N 0.000 claims abstract description 17
- UKLNMMHNWFDKNT-UHFFFAOYSA-M sodium chlorite Chemical compound [Na+].[O-]Cl=O UKLNMMHNWFDKNT-UHFFFAOYSA-M 0.000 claims abstract description 4
- 229960002218 sodium chlorite Drugs 0.000 claims abstract description 4
- 238000003756 stirring Methods 0.000 claims description 24
- -1 chlorite ions Chemical class 0.000 claims description 19
- 239000001963 growth medium Substances 0.000 abstract description 6
- 230000035755 proliferation Effects 0.000 abstract 1
- 238000000034 method Methods 0.000 description 15
- OSVXSBDYLRYLIG-UHFFFAOYSA-N dioxidochlorine(.) Chemical compound O=Cl=O OSVXSBDYLRYLIG-UHFFFAOYSA-N 0.000 description 10
- 239000003814 drug Substances 0.000 description 10
- 230000007704 transition Effects 0.000 description 9
- 229940079593 drug Drugs 0.000 description 6
- 239000004155 Chlorine dioxide Substances 0.000 description 5
- 235000019398 chlorine dioxide Nutrition 0.000 description 5
- 241000195628 Chlorophyta Species 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 230000007423 decrease Effects 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
- 230000000694 effects Effects 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241000224489 Amoeba Species 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000005192 partition Methods 0.000 description 2
- 244000062645 predators Species 0.000 description 2
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- QDHHCQZDFGDHMP-UHFFFAOYSA-N Chloramine Chemical compound ClN QDHHCQZDFGDHMP-UHFFFAOYSA-N 0.000 description 1
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 1
- 241000223782 Ciliophora Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 241000162833 Trebouxiophyceae sp. MBIC11204 Species 0.000 description 1
- 229930003779 Vitamin B12 Natural products 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 229910021529 ammonia Inorganic materials 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
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 229960001701 chloroform Drugs 0.000 description 1
- FDJOLVPMNUYSCM-WZHZPDAFSA-L cobalt(3+);[(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+3].N#[C-].N([C@@H]([C@]1(C)[N-]\C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C(\C)/C1=N/C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C\C1=N\C([C@H](C1(C)C)CCC(N)=O)=C/1C)[C@@H]2CC(N)=O)=C\1[C@]2(C)CCC(=O)NC[C@@H](C)OP([O-])(=O)O[C@H]1[C@@H](O)[C@@H](N2C3=CC(C)=C(C)C=C3N=C2)O[C@@H]1CO FDJOLVPMNUYSCM-WZHZPDAFSA-L 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- VISKNDGJUCDNMS-UHFFFAOYSA-M potassium;chlorite Chemical compound [K+].[O-]Cl=O VISKNDGJUCDNMS-UHFFFAOYSA-M 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
- 238000003753 real-time PCR Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- 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 1
- 235000019163 vitamin B12 Nutrition 0.000 description 1
- 239000011715 vitamin B12 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
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
本発明は、藻類培養方法に関する。 The present invention relates to an algal culture method.
藻類培養槽で藻類を培養するとき（特に開放系の藻類培養槽の場合）、藻類培養槽内で原生生物や真菌類等が発生することがある。特にアメーバ、鞭毛虫、繊毛虫等の原生生物は藻類を捕食する捕食生物である。これらの捕食生物が藻類培養槽内で増殖すると藻類の培養を阻害してしまう。 When culturing algae in an algae culture tank (especially in the case of an open algae culture tank), protists and fungi may be generated in the algae culture tank. In particular, protists such as amoeba, flagellate and ciliate are predators that prey on algae. If these predators grow in the algal culture tank, the culture of the algae is inhibited.
そこで、藻類培養槽内の原生生物を低減する技術が求められる。特許文献１、２には、船舶のバラスト水中の原生生物を死滅させることを目的とする技術が開示されている。 Therefore, a technique for reducing protists in the algal culture tank is required. Patent Documents 1 and 2 disclose techniques aimed at killing protists in ship ballast water.
しかしながら、特許文献１、２記載の技術を藻類の培養に適用すると、原生生物だけではなく、藻類も死滅するか、増殖が大きく阻害されてしまう。本発明は以上の点に鑑みなされたものであり、藻類の増殖を妨げにくく、原生生物を低減できる藻類培養方法を提供することを目的とする。 However, when the techniques described in Patent Documents 1 and 2 are applied to algae culture, not only protists but also algae are killed or their growth is greatly inhibited. The present invention has been made in view of the above points, and an object of the present invention is to provide an algae culture method that can prevent the growth of algae and reduce protists.
本発明の藻類培養方法は、亜塩素酸塩を含む培養液を用いて藻類を培養する。そして、培養液における亜塩素酸イオンの濃度が１．２ｐｐｍ以下であり、少なくとも一時的に、亜塩素酸イオンの濃度が０．４ｐｐｍ以上であることを特徴とする。 In the algal culture method of the present invention, algae are cultured using a culture solution containing chlorite. And the density | concentration of a chlorite ion in a culture solution is 1.2 ppm or less, The density | concentration of a chlorite ion is 0.4 ppm or more at least temporarily.
本発明の藻類培養方法は、藻類の増殖を妨げにくく、且つ原生生物を低減することができる。 The algae culture method of the present invention can hardly inhibit the growth of algae and can reduce protists.
本発明の実施形態を説明する。培養液が含有する亜塩素酸塩としては、例えば、亜塩素酸ナトリウム、亜塩素酸カリウム等を用いることができる。また、培養液は、例えば、安定化二酸化塩素（亜塩素酸ナトリウムと所定の添加物との混合物）を含有することができる。安定化二酸化塩素は、ｐＨの影響を受けにくいため、例えば、酸性条件下で藻類を培養する際に好ましい。 An embodiment of the present invention will be described. As the chlorite contained in the culture solution, for example, sodium chlorite, potassium chlorite and the like can be used. The culture solution can contain, for example, stabilized chlorine dioxide (a mixture of sodium chlorite and a predetermined additive). Stabilized chlorine dioxide is less susceptible to the influence of pH, and is preferable, for example, when culturing algae under acidic conditions.
培養液における亜塩素酸イオンの濃度は、新たに亜塩素酸塩を添加しなければ、時間の経過とともに徐々に減少する。培養液に亜塩素酸塩を添加した場合、培養液における亜塩素酸イオンの濃度は、添加直後に最大となり、時間の経過とともに徐々に減少する。 The concentration of chlorite ions in the culture solution gradually decreases with time unless new chlorite is added. When chlorite is added to the culture solution, the concentration of chlorite ions in the culture solution becomes maximum immediately after the addition, and gradually decreases with time.
なお、本発明における亜塩素酸イオンの濃度とは、培養液全体における平均的な値である。培養液に亜塩素酸塩を添加する場合は、培養液の攪拌を十分に行い、亜塩素酸塩の添加場所から十分離れた位置において亜塩素酸イオンの濃度を測定することが好ましい。 In addition, the density | concentration of chlorite ion in this invention is an average value in the whole culture solution. When chlorite is added to the culture solution, it is preferable to sufficiently stir the culture solution and measure the concentration of chlorite ions at a position sufficiently away from the chlorite addition site.
本発明では、培養液における亜塩素酸イオンの濃度が、（例えば亜塩素酸塩の添加直後においても）、１．２ｐｐｍ以下である。そのことにより、藻類の増殖を妨げにくい。
また、亜塩素酸イオンの濃度が時間の経過とともに徐々に減少したとしても、亜塩素酸イオンの濃度は、少なくとも一時的には０．４ｐｐｍ以上である。そのことにより、原生生物を低減することができる。
In the present invention, the concentration of chlorite ions in the culture solution is 1.2 ppm or less (for example, immediately after addition of chlorite). This makes it difficult to prevent the growth of algae.
Moreover, even if the concentration of chlorite ions gradually decreases with the passage of time, the concentration of chlorite ions is at least temporarily at least 0.4 ppm. As a result, protists can be reduced.
亜塩素酸イオンの濃度が０．４ｐｐｍ以上である時間は、１時間以上（より好ましくは２時間以上）継続することが好ましい。１時間以上継続することで、原生生物を低減する効果が一層高くなる。 The time during which the concentration of chlorite ions is 0.4 ppm or more is preferably continued for 1 hour or more (more preferably 2 hours or more). By continuing for 1 hour or more, the effect of reducing protists is further enhanced.
本発明の藻類培養方法は、例えば、亜塩素酸塩を含む溶液を培養液に添加する工程Ａを有し、その溶液の添加中は培養液を攪拌し、溶液の添加後は培養液の攪拌を停止することができる。 The algae culture method of the present invention has, for example, the step A of adding a solution containing chlorite to the culture solution, stirring the culture solution during the addition of the solution, and stirring the culture solution after the addition of the solution. Can be stopped.
この場合、溶液の添加中は攪拌を行うので、局所的に亜塩素酸イオン濃度が高い部分が培養液中に生じにくい。また、溶液の添加後、攪拌を停止するので、亜塩素酸イオン濃度の低下を抑制し、亜塩素酸イオン濃度が０．４ｐｐｍ以上である時間を延ばすことができる。 In this case, since stirring is performed during the addition of the solution, a portion having a locally high chlorite ion concentration is unlikely to occur in the culture solution. Moreover, since stirring is stopped after addition of a solution, the fall of a chlorite ion density | concentration can be suppressed and the time when a chlorite ion density | concentration is 0.4 ppm or more can be extended.
培養液の攪拌を停止した後、所定時間が経過すれば、攪拌を再開してもよい。この場合、藻類の培養を一層適切に行うことができる。
亜塩素酸塩を含む溶液を培養液に添加する場合、その溶液における亜塩素酸イオンの濃度は、２．５〜１０ｐｐｍであることが好ましい。１０ｐｐｍ以下であることにより、局所的に亜塩素酸イオン濃度が高い部分が培養液中に生じにくい。また、２．５ｐｐｍ以上であることにより、培養液における亜塩素酸イオン濃度を所望の濃度まで速やかに高めることができる。
Stirring may be resumed if a predetermined time elapses after stirring of the culture broth is stopped. In this case, algae can be cultured more appropriately.
When a solution containing chlorite is added to the culture solution, the concentration of chlorite ions in the solution is preferably 2.5 to 10 ppm. By being 10 ppm or less, a portion having a locally high chlorite ion concentration is unlikely to occur in the culture solution. Moreover, the chlorite ion density | concentration in a culture solution can be rapidly raised to a desired density | concentration because it is 2.5 ppm or more.
本発明の藻類培養方法は、例えば、培養液の一部及びそれに含まれる藻類を取り出す工程Ｂを有し、前記工程Ａは、前記工程Ｂの後で行うことができる。この場合、工程Ｂの後における培養液の補充と、亜塩素酸塩の添加とを同時に行うことができる。 The algae culture method of the present invention includes, for example, a step B for removing a part of the culture solution and the algae contained therein, and the step A can be performed after the step B. In this case, replenishment of the culture solution after step B and addition of chlorite can be performed simultaneously.
培養する藻類としては、例えば、緑藻植物が挙げられる。緑藻植物としては、例えば、トレボウクシア藻綱や緑藻網等が挙げられる。トレボウクシア藻綱としては、例えば、クロレラ目、トレボウクシア目等が挙げられる。トレボウクシア目としては、例えば、シュードコリシスティス（Pseudochoricystis ellipsoidea）等が挙げられる。 Examples of the algae to be cultured include green algae plants. Examples of green algae plants include Trevoxia algae and green algae net. Examples of Trevoxia algae include Chlorella, Trevoxia and the like. Examples of Trevoxia include Pseudochoricystis ellipsoidea and the like.
本発明の藻類培養方法により低減できる原生生物として、例えば、アメーバ、鞭毛虫、真菌類等の生物、及びこれらの生物の増殖因子である細菌等が挙げられる。
＜実施例＞
１．藻類の培養に用いる培養システム１の構成
藻類の培養に用いる培養システム１の構成を、図１及び図２に基いて説明する。培養システム１は、培養槽３と、薬剤供給機構５とを備える。培養槽３はいわゆるレースウェイ型の槽であり、中央の仕切板７、及び攪拌用パドル９を備えており、攪拌用パドル９の回転により、培養槽３内の培養液４に、仕切板７の回りを周回する、図１に示す矢印Ｆ方向の流れを生じさせることができる。
Examples of protists that can be reduced by the algal culture method of the present invention include organisms such as amoeba, flagellate, and fungi, and bacteria that are growth factors of these organisms.
<Example>
1. Configuration of Culture System 1 Used for Algal Culture The configuration of the culture system 1 used for algae culture will be described with reference to FIGS. 1 and 2. The culture system 1 includes a culture tank 3 and a drug supply mechanism 5. The culture tank 3 is a so-called raceway-type tank, and is provided with a central partition plate 7 and a stirring paddle 9. The flow in the direction of arrow F shown in FIG.
薬剤供給機構５は、薬剤を保持するタンク１１と、薬剤を培養槽３に供給する配管１３とを備える。配管１３は、その一方の端部１３Ａにおいてタンク１１の内部と連通するとともに、その反対側の端部１３Ｂ側を培養槽３上に張り出している。端部１３Ｂの位置は、攪拌用パドル９の近傍であって、培養槽３における流れ方向に関し、攪拌用パドル９よりも上流側である。 The drug supply mechanism 5 includes a tank 11 that holds a drug and a pipe 13 that supplies the drug to the culture tank 3. The pipe 13 communicates with the inside of the tank 11 at one end 13 </ b> A, and the opposite end 13 </ b> B side projects over the culture tank 3. The position of the end portion 13B is in the vicinity of the stirring paddle 9 and upstream of the stirring paddle 9 with respect to the flow direction in the culture tank 3.
図２に示すように、配管１３の下面（図２における下側の面）のうち、端部１３Ｂの近傍には、複数の孔１５が設けられている。薬剤供給機構５は、タンク１１内に保持する薬剤を、図示しないポンプによって配管１３内に送り出すことができる。配管１３内に送り出された薬剤は、複数の孔１５を通って落下し、培養槽３内の培養液４に添加される。薬剤が落下する位置は、攪拌用パドル９の近傍であって、培養槽３における流れ方向に関し、攪拌用パドル９よりも上流側の位置である。 As shown in FIG. 2, a plurality of holes 15 are provided in the vicinity of the end 13 </ b> B on the lower surface of the pipe 13 (the lower surface in FIG. 2). The medicine supply mechanism 5 can send out the medicine held in the tank 11 into the pipe 13 by a pump (not shown). The medicine sent into the pipe 13 falls through the plurality of holes 15 and is added to the culture solution 4 in the culture tank 3. The position where the medicine falls is in the vicinity of the stirring paddle 9 and on the upstream side of the stirring paddle 9 with respect to the flow direction in the culture tank 3.
２．藻類の培養方法
以下の方法で藻類の培養を行った。
（２−１）培養槽３に５００Ｌの培養液と藻類とを入れ、藻類の培養を行った。培養液は周知のＡＦ６培地と同様であり、その１００ｍｌにおける組成は以下のとおりである。塩酸、硫酸等を用いて、藻類培養開始時の培養液ｐＨを４に調整し、培養中はｐＨ３〜４に保持した。使用する安定化二酸化塩素はダイソー社製のオスロン（商品名）である。
2. Algal culture method Algal culture was performed by the following method.
(2-1) A 500 L culture solution and algae were placed in the culture tank 3, and algae were cultured. The culture solution is similar to the well-known AF6 medium, and its composition in 100 ml is as follows. Using hydrochloric acid, sulfuric acid or the like, the culture solution pH at the start of algal culture was adjusted to 4 and maintained at pH 3 to 4 during the culture. The stabilized chlorine dioxide to be used is Oslon (trade name) manufactured by Daiso Corporation.
NaNO3：14 mg
NH4NO3：2.2 mg
MgSO4 ・ 7H2O：3 mg
KH2PO4：1 mg
K2HPO4：0.5 mg
CaCl2 ・ 2H2O ：1 mg
CaCO3：1 mg
Fe-citrate：0.2 mg
Citric acid ：0.2 mg
Biotin：0.2 μg
Thiamine HCl：1 μg
Vitamin B6：0.1 μg
Vitamin B12：0.1 μg
Trace metals：0.5 mL
Distilled water：99.5 mL
(pH 6.62)
また、培養した藻類は、緑藻植物であって、トレボウクシア藻綱、トレボウクシア目のシュードコリシスティスMBIC11204株である。培養中は、攪拌用パドル９を継続的に回転させ、培養液４に、図１に示す矢印Ｆ方向の流れを生じさせた。
NaNO 3 : 14 mg
NH 4 NO 3 : 2.2 mg
MgSO 4・ 7H 2 O ： 3 mg
KH 2 PO 4 : 1 mg
K 2 HPO 4 : 0.5 mg
CaCl 2 · 2H 2 O: 1 mg
CaCO 3 : 1 mg
Fe-citrate: 0.2 mg
Citric acid: 0.2 mg
Biotin: 0.2 μg
Thiamine HCl: 1 μg
Vitamin B6: 0.1 μg
Vitamin B12: 0.1 μg
Trace metals: 0.5 mL
Distilled water: 99.5 mL
(pH 6.62)
The cultured algae are green algae plants, such as Trevoxia algae, Trevoxia pseudocolistis strain MBIC11204. During the culture, the stirring paddle 9 was continuously rotated to cause the culture solution 4 to flow in the direction of arrow F shown in FIG.
培養液中の藻類の濃度を定期的に測定しながら、藻類の濃度が０．２μｇ／Ｌとなるまで培養を続けた。なお、培養液中の藻類の濃度は、所定量の培養液を測り取り、その培養液を濾過して藻類を分離し、分離した藻類の乾燥重量を測定することで算出した。
（２−２）培養液中の藻類の濃度が０．２μｇ／Ｌに達すると、培養槽３内の培養液の半分（２５０Ｌ）を、その中に藻類を含んだ状態のまま、取り出した。なお、上記の工程は工程Ｂの一実施形態である。
（２−３）次に、薬剤供給機構５を用いて、安定化二酸化塩素を含む培養液（以下、添加培養液とする）を、培養槽３内に残った培養液に添加した。添加培養液における亜塩素酸イオン濃度は４ｐｐｍである。また、添加培養液における安定化二酸化塩素以外の成分は、周知のＡＦ６培地と同様である。
The culture was continued until the algae concentration reached 0.2 μg / L while periodically measuring the algae concentration in the culture solution. The concentration of algae in the culture solution was calculated by measuring a predetermined amount of the culture solution, filtering the culture solution to separate the algae, and measuring the dry weight of the separated algae.
(2-2) When the concentration of algae in the culture solution reached 0.2 μg / L, half (250 L) of the culture solution in the culture tank 3 was taken out while containing the algae therein. In addition, said process is one Embodiment of the process B.
(2-3) Next, using the drug supply mechanism 5, a culture solution containing stabilized chlorine dioxide (hereinafter referred to as an added culture solution) was added to the culture solution remaining in the culture tank 3. The concentration of chlorite ion in the added culture medium is 4 ppm. Moreover, components other than the stabilized chlorine dioxide in the added culture medium are the same as those in the well-known AF6 medium.
添加培養液の添加量は、培養槽３内の培養液において亜塩素酸イオンの濃度が初期濃度Ｃ０となる量とした。また、添加培養液の添加速度は、２〜８Ｌ／ｍｉｎとした。また、添加培養液を添加している間は、攪拌用パドル９を引き続き回転させ、培養液を攪拌した。 The addition amount of the supplemented culture solution, the concentration of chlorite ion is an amount that the initial concentration C 0 in the culture solution in the culture vessel 3. Moreover, the addition speed | rate of the addition culture solution was 2-8 L / min. Further, while the added culture solution was being added, the stirring paddle 9 was continuously rotated to stir the culture solution.
なお、上記の工程は工程Ａの一実施形態であり、添加培養液は、亜塩素酸塩を含む溶液の一実施形態である。
（２−４）必要な量の添加培養液を添加し終わると、攪拌用パドル９の回転を止めて培養液の攪拌を停止し、１時間静置した。一時間の静置が終了すると、培養槽３内に水を加え、培養槽３内の液量を５００Ｌに戻した。
In addition, said process is one Embodiment of process A, and an addition culture solution is one Embodiment of the solution containing a chlorite.
(2-4) When the required amount of added culture broth was added, the rotation of the stirring paddle 9 was stopped, stirring of the culture broth was stopped, and the culture was allowed to stand for 1 hour. When the standing for 1 hour was completed, water was added to the culture tank 3, and the amount of liquid in the culture tank 3 was returned to 500L.
また、上記の一時間の静置が収量すると、攪拌用パドル９の回転を再開し、培養液の攪拌を行った。そして、培養液中の藻類の濃度を定期的に測定しながら、藻類の濃度が０．２μｇ／Ｌとなるまで培養を続けた。 Further, when the above-mentioned standing for 1 hour yielded, the rotation of the stirring paddle 9 was resumed, and the culture solution was stirred. And culture | cultivation was continued until the density | concentration of algae became 0.2 microgram / L, measuring the density | concentration of the algae in a culture solution regularly.
以降、上記（２−２）〜（２−４）の工程を繰り返し、藻類の培養を行った。
上記の培養方法であって、初期濃度Ｃ０が０ｐｐｍの場合（コントロール）、０．５ｐｐｍの場合、１．１ｐｐｍの場合、及び１．５ｐｐｍの場合をそれぞれ行った。そして、それぞれの条件での培養中に、培養槽３内の培養液における亜塩素酸イオンの濃度と、藻類の濃度とを繰り返し測定した。その結果を図３〜図５に示す。また図６に、初期濃度Ｃ０が０ｐｐｍの場合（コントロール）、１．１ｐｐｍの場合の原生生物量を示す。
Thereafter, the above steps (2-2) to (2-4) were repeated to culture algae.
In the above culture method, the initial concentration C 0 was 0 ppm (control), 0.5 ppm, 1.1 ppm, and 1.5 ppm. And the density | concentration of the chlorite ion in the culture solution in the culture tank 3 and the density | concentration of algae were repeatedly measured during culture | cultivation on each conditions. The results are shown in FIGS. FIG. 6 shows the amount of protists when the initial concentration C 0 is 0 ppm (control) and 1.1 ppm.
なお、図３〜６における横軸（時間軸）の０は、添加培養液の添加を開始した時点を意味する。亜塩素酸イオン濃度は、デジタル残留塩素テスターDCT-05（タクミナ製）を用いて測定した。また、藻類の濃度は上述した方法で測定した。また、原生生物の量は、定量ＰＣＲの方法で測定し、ＡＢＩ社製の測定装置であるStepOnePlus（商品名）を使用した。図６の縦軸は、添加培養液の添加開始時における原生生物の量を１とした場合の相対値である。 In addition, 0 of the horizontal axis (time axis) in FIGS. 3-6 means the time of starting the addition of an addition culture solution. The chlorite ion concentration was measured using a digital residual chlorine tester DCT-05 (manufactured by Takumina). Moreover, the density | concentration of algae was measured by the method mentioned above. In addition, the amount of protists was measured by a quantitative PCR method, and StepOnePlus (trade name), which is a measuring device manufactured by ABI, was used. The vertical axis in FIG. 6 is a relative value when the amount of protists at the start of addition of the added culture solution is 1.
図３Ａに示すように、初期濃度Ｃ０が０．５ｐｐｍの場合、亜塩素酸イオンの濃度は１．２ｐｐｍ以下であり、また、亜塩素酸イオンの濃度が０．４ｐｐｍ以上である時間が１時間以上継続していた。そして、図３Ｂに示すように、初期濃度Ｃ０が０．５ｐｐｍの場合の藻類濃度の推移は、コントロールの場合とほとんど変わらなかった。すなわち、亜塩素酸イオンにより、藻類の増殖が阻害されることはなかった。 As shown in FIG. 3A, when the initial concentration C 0 is 0.5 ppm, the concentration of chlorite ions is 1.2 ppm or less, and the time during which the concentration of chlorite ions is 0.4 ppm or more is 1 It continued for more than an hour. Then, as shown in FIG. 3B, transition initial concentration C 0 of algae concentration in the case of 0.5ppm is hardly changed in the case of control. That is, the growth of algae was not inhibited by chlorite ions.
また、図４Ａに示すように、初期濃度Ｃ０が１．１ｐｐｍの場合、亜塩素酸イオンの濃度は１．２ｐｐｍ以下であり、また、亜塩素酸イオンの濃度が０．４ｐｐｍ以上である時間が１時間以上継続していた。そして、図４Ｂに示すように、初期濃度Ｃ０が１．１ｐｐｍの場合の藻類濃度の推移は、コントロールの場合とほとんど変わらなかった。すなわち、亜塩素酸イオンにより、藻類の増殖が阻害されることはなかった。 As shown in FIG. 4A, when the initial concentration C 0 is 1.1 ppm, the concentration of chlorite ions is 1.2 ppm or less, and the time during which the concentration of chlorite ions is 0.4 ppm or more. Continued for over an hour. Then, as shown in FIG. 4B, transition initial concentration C 0 of algae concentration in the case of 1.1ppm is hardly changed in the case of control. That is, the growth of algae was not inhibited by chlorite ions.
一方、図５Ａに示すように、初期濃度Ｃ０が１．５ｐｐｍの場合、亜塩素酸イオンの濃度は１．２ｐｐｍを超えていた。そして、図５Ｂに示すように、初期濃度Ｃ０が１．５ｐｐｍの場合の藻類濃度は、コントロールの場合に比べて、大きく低下していた。すなわち、亜塩素酸イオンにより、藻類の増殖が阻害されていた。 On the other hand, as shown in FIG. 5A, when the initial concentration C 0 was 1.5 ppm, the concentration of chlorite ions exceeded 1.2 ppm. Then, as shown in FIG. 5B, algae concentration when the initial concentration C 0 is 1.5ppm, compared to the case of the control, it was reduced significantly. That is, the growth of algae was inhibited by chlorite ions.
また、図６に示すように、初期濃度Ｃ０が１．１ｐｐｍの場合、コントロールに比べて、原生生物の量が大きく低下していた。すなわち、亜塩素酸イオンにより、原生生物を減少させることができた。 Further, as shown in FIG. 6, when the initial concentration C 0 was 1.1 ppm, the amount of protists was greatly reduced compared to the control. In other words, protists could be reduced by chlorite ions.
３．藻類の培養方法が奏する効果
（１）本実施例の培養方法（初期濃度Ｃ０が０．５ｐｐｍ、１．１ｐｐｍの場合）は、藻類の増殖を妨げにくく、且つ原生生物を低減することができる。
3. Culturing method Effect (1) In this embodiment the culture process is achieved algae (when the initial concentration C 0 is 0.5 ppm, of 1.1 ppm) is unlikely to interfere with the growth of algae, and can be reduced protists .
（２）本実施例の培養方法では、添加培養液の添加中は培養液を攪拌するので、局所的に亜塩素酸イオン濃度が高い部分が培養液中に生じにくい。そのため、藻類の増殖を一層促進することができる。 (2) In the culture method of the present example, the culture solution is stirred during the addition of the added culture solution, so that a portion having a locally high chlorite ion concentration is unlikely to occur in the culture solution. Therefore, the growth of algae can be further promoted.
また、本実施例の培養方法では、添加培養液の添加後、攪拌を停止するので、亜塩素酸イオン濃度の低下を抑制し、亜塩素酸イオン濃度が０．４ｐｐｍ以上である時間を延ばすことができる。そのため、原生生物を一層低減することができる。 In addition, in the culture method of this example, the stirring is stopped after the addition of the added culture solution, so that the decrease in the chlorite ion concentration is suppressed and the time during which the chlorite ion concentration is 0.4 ppm or more is extended. Can do. Therefore, protists can be further reduced.
（３）本実施例の培養方法では、添加培養液における亜塩素酸イオンの濃度が４ｐｐｍである。そのため、添加培養液の添加中に、局所的に亜塩素酸イオン濃度が高い部分が培養液中に生じにくい。また、培養液における亜塩素酸イオン濃度を所望の初期濃度Ｃ０まで速やかに高めることができる。 (3) In the culture method of the present example, the concentration of chlorite ions in the added culture solution is 4 ppm. For this reason, a portion having a locally high chlorite ion concentration is unlikely to occur in the culture solution during the addition of the added culture solution. Further, it is possible to increase rapidly the chlorite ion concentration in the culture medium to the desired initial concentration C 0.
（４）本実施例の培養方法では、培養液の半分を取り出す工程の後で、添加培養液を添加する。そのため、培養槽３に対する培養液の補充と、亜塩素酸塩の添加とを同時に行うことができる。 (4) In the culture method of the present embodiment, the added culture solution is added after the step of taking out half of the culture solution. Therefore, replenishment of the culture solution to the culture tank 3 and addition of chlorite can be performed simultaneously.
（５）本実施例の培養方法では、添加培養液の添加速度が２〜８Ｌ／ｍｉｎである。そのため、添加培養液の添加中に、局所的に亜塩素酸イオン濃度が高い部分が培養液中に生じにくい。 (5) In the culture method of the present example, the addition rate of the added culture solution is 2 to 8 L / min. For this reason, a portion having a locally high chlorite ion concentration is unlikely to occur in the culture solution during the addition of the added culture solution.
（６）本実施例の培養方法では、添加培養液を添加する位置が、攪拌用パドル９の近傍であって、培養槽３における流れ方向に関し、攪拌用パドル９よりも上流側の位置である。そのため、添加された添加培養液は攪拌用パドル９により速やかに攪拌されるので、局所的に亜塩素酸イオン濃度が高い部分が培養液中に生じにくい。 (6) In the culturing method of the present embodiment, the position where the added culture solution is added is in the vicinity of the stirring paddle 9 and is upstream of the stirring paddle 9 in the flow direction in the culture tank 3. . Therefore, since the added culture medium is rapidly stirred by the paddle 9 for stirring, a portion having a locally high chlorite ion concentration is unlikely to occur in the culture liquid.
（７）特許文献１、２記載の技術で使用する次亜塩素酸ナトリウムはアンモニアや有機物と反応して有害なクロラミンやトリクロロメタンを生成するが、本実施形態の培養方法では、次亜塩素酸ナトリウムを使用しないので、そのような問題が生じない。 (7) Sodium hypochlorite used in the techniques described in Patent Documents 1 and 2 reacts with ammonia and organic substances to produce harmful chloramine and trichloromethane. In the culture method of this embodiment, hypochlorous acid is used. Since no sodium is used, such a problem does not occur.
また、特許文献１、２記載の技術では、ｐＨ５以下の場合、急速に分解反応が起こり、効果が減少してしまうが、本実施形態の培養方法では、そのような問題が生じにくい。 Further, in the techniques described in Patent Documents 1 and 2, when the pH is 5 or less, a decomposition reaction occurs rapidly and the effect is reduced. However, in the culture method of the present embodiment, such a problem hardly occurs.
１…培養システム、３…培養槽、４…培養液、５…薬剤供給機構、７…仕切板、９…攪拌用パドル、１１…タンク、１３…配管、１３Ａ、１３Ｂ…端部、１５…孔 DESCRIPTION OF SYMBOLS 1 ... Culture system, 3 ... Culture tank, 4 ... Culture solution, 5 ... Drug supply mechanism, 7 ... Partition plate, 9 ... Paddle for stirring, 11 ... Tank, 13 ... Pipe, 13A, 13B ... End part, 15 ... Hole
Claims (6)
前記培養液における亜塩素酸イオンの濃度が１．２ｐｐｍ以下であり、
少なくとも一時的に、前記濃度が０．４ｐｐｍ以上であることを特徴とする藻類培養方法。 Using a culture solution containing chlorite,
The concentration of chlorite ions in the culture broth is 1.2 ppm or less,
At least temporarily, the said density | concentration is 0.4 ppm or more, The algae culture method characterized by the above-mentioned.
前記溶液の添加中は前記培養液を攪拌し、前記溶液の添加後、前記培養液の攪拌を停止することを特徴とする請求項１〜３のいずれか１項に記載の藻類培養方法。 Adding the solution containing the chlorite to the culture solution A,
The algae culture method according to any one of claims 1 to 3, wherein the culture solution is stirred during the addition of the solution, and the stirring of the culture solution is stopped after the addition of the solution.
前記工程Ａは、前記工程Ｂの後で行われることを特徴とする請求項４又は５に記載の藻類培養方法。 A step B of taking out a part of the culture solution and the algae contained therein,
6. The algae culture method according to claim 4 or 5, wherein the step A is performed after the step B.
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