CN1504109A - Liquid containing diatom, diatom and method for culturing diatom - Google Patents

Liquid containing diatom, diatom and method for culturing diatom Download PDF

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CN1504109A
CN1504109A CNA200310120738A CN200310120738A CN1504109A CN 1504109 A CN1504109 A CN 1504109A CN A200310120738 A CNA200310120738 A CN A200310120738A CN 200310120738 A CN200310120738 A CN 200310120738A CN 1504109 A CN1504109 A CN 1504109A
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diatom
nitrogen
silicon
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liquid
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CN100500828C (en
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近藤裕
郎
山内一郎
士彦
富田富士彦
子
柏仓真
山下卓
铃木顺子
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Japanese Polytron Technologies Inc Marin
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Nisshin Oillio Ltd
Yamaha Motor Co Ltd
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Abstract

A liquid for culturing diatom comprising nitrogen, silicon and phosphorus, wherein a value of silicon/nitrogen, representing the ratio of concentration by mass of silicon to nitrogen, is set to be a value of 0.06 or more, and a value of phosphorus/nitrogen, representing the ratio of concentration by mass of phosphorus to nitrogen, is set to be a value of 0.18 or more, and the pH of the culture liquid is set in the range of 6.4 to 8.4. The culture liquid allows the culture of Chaetoceros at a cell concentration greater than the maximum value (3 X 10<13> cells/m<3>) achieved by the conventional culture methods and with stability.

Description

The cultural method of diatomaceous liquid, diatom and diatom
Technical field
The present invention relates to diatom as microalgae, described microalgae is used for shellfishes such as oyster, Margarita, and japonicus, base enclose crustaceans such as shrimp, the bait of aquatic products seedlings such as Echinodermata such as sea cucumber, sea urchin, more specifically, relate to the liquid that contains diatom and the cultural method of diatom.
Background technology
Bait during as aquatic products seedlings such as cultivated shellfish, crustaceans, Echinodermatas, in recent years, the diatom (for example calcareous Chaetoceros (calcitrans) that belongs to of Chaetoceros, angle thorn algae (gracilis) etc.) that is categorized as microalgaes such as Chaetoceros (Chaetoceros) genus, brown algae (Phaeodactylum) genus has begun to be paid close attention to by people.
For example, calcareous Chaetoceros has 1 long flagellum respectively on 4 angles.Because the diameter (total length) of calcareous Chaetoceros is about about 3～6 μ m, the bait when therefore being especially suitable for use as aquatic products seedling seedling.
As the diatom of one of microalgae, compare with the other plant planktonic organism, good aspect mineral equilibrium and VITAMIN balance.In addition, diatom contains EPA (timnodonic acid, polybasic unsaturated fatty acid), DHA useful lipid acid such as (docosahexenoic acid, polybasic unsaturated fatty acids).For the foregoing reasons, the diatom in the microalgae is obtaining very high evaluation aspect bait.
But the diatom in the microalgae extremely difficulty is stably cultivated.In [1 fish multiplication engineering test, (1) salt solution seedling production research, biological feed is cultivated] in (being designated hereinafter simply as document 1) and [putting down into a large amount of culture technique development research reports of 4 years particular studies exploitation promotion cause microalgaes] (being designated hereinafter simply as document 2), reported that the cell concn upper limit of the diatom Chaetoceros (hereinafter to be referred as Chaetoceros) that obtains by existing cultural method is about 3.0 * 10 13Cell/m 3Cell concn is represented every 1m herein 3Cell count in the nutrient solution.
By existing cultural method, because the cell concn upper limit of Chaetoceros is about 3.0 * 10 13Cell/m 3, therefore can't obtain sufficient cultivation amount.Therefore seeking can high density and stably cultivate the liquid (nutrient solution, substratum) and the cultural method of Chaetoceros.
Summary of the invention
The present invention finishes in view of the above problems, its purpose be to provide a kind of can high density and stably cultivate liquid that contains diatom and diatom cultural method as the diatom of one of microalgae, and provide a kind of diatom of cultivating as stated above.
To achieve these goals, the invention provides a kind of liquid that contains diatom, is the liquid that is used to cultivate diatom, it is characterized in that described liquid contains diatom, silicon and nitrogen, and silicon/nitrogen (mass concentration ratio of described silicon and described nitrogen) is set at and surpasses 0.18 value.
To achieve these goals, the invention provides a kind of diatom, it is characterized in that, described diatom is cultivated in following liquid and forms, and described liquid contains silicon and nitrogen, and silicon/nitrogen (mass concentration ratio of described silicon and described nitrogen) is set at and surpasses 0.18 value.
According to the present invention, be set at and surpass 0.18 value owing to will be used for cultivating the silicon/nitrogen (mass concentration ratio of described silicon and described nitrogen) of the liquid of diatom, so can high density and stably cultivate diatom.
To achieve these goals, the invention provides a kind of liquid that contains diatom, be the liquid that is used to cultivate diatom, it is characterized in that, described liquid contains diatom, phosphorus, silicon and nitrogen, phosphorus/nitrogen (mass concentration ratio of described phosphorus and described nitrogen), and silicon/nitrogen (mass concentration ratio of described silicon and described nitrogen) be set at value more than 0.1 or 0.1 respectively.
To achieve these goals, the invention provides a kind of diatom, it is characterized in that, described diatom is cultivated in following liquid and forms, described liquid contains phosphorus, nitrogen and silicon, and phosphorus/nitrogen (mass concentration ratio of described phosphorus and described nitrogen), and silicon/nitrogen (mass concentration ratio of described silicon and described nitrogen) be set at respectively more than 0.1 or 0.1.
According to the present invention and since will be used for cultivating diatom liquid institute phosphorous/nitrogen (mass concentration ratio of phosphorus and nitrogen), and silicon/nitrogen (mass concentration ratio of silicon and nitrogen) be set at value more than 0.1 or 0.1 respectively, so can high density and stably cultivate diatom.
In order to realize purpose of the present invention, the invention provides a kind of liquid that contains diatom, be the liquid that is used to cultivate diatom, it is characterized in that described liquid contains diatom, phosphorus, silicon and nitrogen, pH is more than 6.4 or 6.4, below 8.4 or 8.4.
In order to realize purpose of the present invention, the invention provides a kind of diatom, it is characterized in that described diatom is cultivated and forms in following liquid, described liquid contains phosphorus, nitrogen and silicon, and pH is set in the scope more than 6.4 or 6.4, below 8.4 or 8.4.
According to the present invention, be set in the scope 6.4 or 6.4 or more, below 8.4 or 8.4 owing to will be used to cultivate the pH of the liquid of diatom, so can high density and stably cultivate diatom.
To achieve these goals, the invention provides the cultural method of a kind of diatom, be to use the diatom nutrient solution that contains nitrogen and silicon to cultivate the diatom cultural method of diatom, it is characterized in that, described method comprise the steps: to measure respectively in the described liquid the determination step in advance of initial stage concentration of nitrogenous initial stage concentration and silicon; The inoculation step of the described diatom of inoculation in described liquid; Measure determination step before the supply of concentration before the supply of concentration and described silicon before the supply of described nitrogen respectively; Obtain the concentration difference determination step of nitrogen concentration poor (concentration is poor before the initial stage concentration of described nitrogen and the supply) and silicon concentration poor (concentration is poor before the initial stage concentration of described silicon and the supply) respectively; Poor based on described nitrogen concentration difference and described silicon concentration, determine the nitrogen amount and the silicon amount of supply in described liquid respectively, and described liquid gone in its supply so that with the consistent supply step of ratio of described nitrogen initial stage concentration and described silicon initial stage concentration.
According to the present invention, before the inoculation diatom, measure the initial stage concentration of nitrogen and the initial stage concentration of silicon, before supply nitrogen and silicon, measure the preceding concentration of supply of nitrogen and the preceding concentration of supply of silicon.Then, based on nitrogen concentration poor (concentration is poor before the initial stage concentration of nitrogen and the supply) and silicon concentration poor (concentration is poor before the initial stage concentration of silicon and the supply), the nitrogen amount and the silicon amount of supply in the decision liquid.Thereby, because the ratio of nitrogen concentration and siliceous amount concentration is consistent, so can high density and stably cultivate diatom.
To achieve these goals, the invention provides the cultural method of a kind of diatom, be to use the diatom nutrient solution that contains nitrogen and silicon to cultivate the diatom cultural method of diatom, it is characterized in that described method comprises the steps: to measure respectively determination step before the supply of concentration before the supply of concentration and described silicon before the supply of described nitrogen; Obtain the concentration difference determination step of nitrogen concentration poor (concentration is poor before the normality of described nitrogen and the supply) and silicon concentration poor (concentration is poor before the normality of described silicon and the supply) respectively; Poor based on described nitrogen concentration difference and described silicon concentration, determine the nitrogen amount and the silicon amount of supply in described liquid respectively, and described liquid gone in its supply so that with the consistent supply step of ratio of the normality of the normality of described nitrogen and described silicon.
According to the present invention, before supply nitrogen and silicon, after the preceding concentration of supply of preceding concentration of the supply of mensuration nitrogen and silicon, based on nitrogen concentration poor (concentration is poor before the normality of nitrogen and the supply) and silicon concentration poor (concentration is poor before the normality of silicon and the supply), the nitrogen amount and the silicon amount of decision supply in liquid.Thereby, because the mass concentration of the nitrogen ratio with the mass concentration of silicon is consistent, so can high density and stably cultivate diatom.
To achieve these goals, the invention provides the cultural method of a kind of diatom, be to use the diatom nutrient solution that contains nitrogen and silicon to cultivate the diatom cultural method of diatom, it is characterized in that, described method comprise the steps: to measure in the described liquid of inoculating diatom the determination step in advance of initial stage concentration of nitrogenous initial stage concentration and silicon; Measure determination step before the supply of concentration before the supply of concentration and described silicon before the supply of described nitrogen respectively; Obtain the concentration difference determination step of nitrogen concentration poor (concentration is poor before the initial stage concentration of described nitrogen and the supply) and silicon concentration poor (concentration is poor before the initial stage concentration of described silicon and the supply) respectively; Poor based on described nitrogen concentration difference and described silicon concentration, determine the nitrogen amount and the silicon amount of supply in described liquid respectively, and its supply gone in the described liquid so that with the consistent supply step of ratio of the initial stage concentration of the initial stage concentration of described nitrogen and described silicon.
According to the present invention, behind the inoculation diatom, measure the initial stage concentration of nitrogen and the initial stage concentration of silicon, before supply nitrogen and silicon, measure the preceding concentration of supply of nitrogen and the preceding concentration of supply of silicon.Then, based on nitrogen concentration poor (concentration is poor before the initial stage concentration of described nitrogen and the supply) and silicon concentration poor (concentration is poor before the initial stage concentration of described silicon and the supply), the nitrogen amount and the silicon amount of supply in the decision liquid.Thereby, because the mass concentration of the nitrogen ratio with the mass concentration of silicon is consistent, so can high density and stably cultivate diatom.
To achieve these goals, the invention provides the cultural method of a kind of diatom, be to use the nutrient solution that contains nitrogen and silicon to cultivate the diatom cultural method of diatom, it is characterized in that, described method comprises the steps: to measure with in the liquid at the consumption of having inoculated described diatom, measures the nitrogen consumption of described diatom consumption and the determination step in advance of silicon consumption respectively; The inoculation step of the described diatom of inoculation in described nutrient solution; Measure the determination step of the cell concn of described diatom; Described cell concn surpasses under the situation of specified amount, and modulation is topdressed and used liquid, the modulation step that the ratio of the ratio of nitrogen and silicon and described nitrogen consumption and described silicon consumption roughly is consistent; Synthetic described topdressing gone into supply step in the described nutrient solution with the liquid supply.
According to the present invention, measure for liquid for consumption, behind the nitrogen consumption and silicon consumption measured respectively by diatom consumption, when cultivating diatom with nutrient solution, if the cell concn of diatom surpasses specified amount, then the ratio decision with nitrogen and silicon roughly is consistent for the ratio with nitrogen consumption and silicon consumption.Thereby, because the mass concentration of the nitrogen ratio with the mass concentration of silicon is remained necessarily, so can high density and stably cultivate diatom.
Description of drawings
Fig. 1 illustrates the nutrient solution composition that uses in the present embodiment.
Fig. 2 illustrates cell concn variation, nitrogen consumption, phosphorus consumption and the silicon consumption of the Chaetoceros of breeding in each nutrient solution of sample 1 to 11.
Fig. 3 A illustrates the relation of nitrogen consumption and cell concn variation.
Fig. 3 B illustrates the relation of phosphorus consumption and cell concn variation.
Fig. 3 C illustrates the relation of silicon consumption and cell concn variation.
The one-tenth that Fig. 4 illustrates the nutrient solution 1 to 6 that uses in the present embodiment respectively is grouped into and the one-tenth of existing nutrient solution 7 to 8 is grouped into.
Fig. 5 illustrate in each nutrient solution shown in Figure 4 nitrogenous mass concentration, the mass concentration of phosphorus and the mass concentration of silicon.
Fig. 6 illustrates the pH value of nutrient solution 4 and the relation of amount of hydrolysis.
Fig. 7 is the side elevational view of the flat culturing bottle that uses in the present embodiment.
Fig. 8 is illustrated in pH value that reaches nutrient solution 1 when adding sodium bicarbonate in nutrient solution 1 when adding sodium bicarbonate in the nutrient solution 1 and the relation that cell concn changes respectively.
The mass concentration that Fig. 9 is illustrated in sodium bicarbonate respectively is adjusted into 0.02 * 10 3Ppm, 0.14 * 10 3Ppm, 1.0 * 10 3Cell concn over time in the nutrient solution 1 of ppm.
The mass concentration that Figure 10 is illustrated in sodium bicarbonate respectively is adjusted into 0.02 * 10 3Ppm, 0.14 * 10 3Ppm, 1.0 * 10 3The pH value over time in the nutrient solution 1 of ppm.
The mass concentration that Figure 11 is illustrated in sodium bicarbonate respectively is adjusted into 0.07 * 10 3Ppm, 0.14 * 10 3Ppm, 0.28 * 10 3Ppm, 1.0 * 10 3Cell concn over time in the nutrient solution 1 of ppm.
The mass concentration that Figure 12 is illustrated in sodium bicarbonate respectively is adjusted into 0.07 * 10 3Ppm, 0.14 * 10 3Ppm, 0.28 * 10 3Ppm, 1.0 * 10 3The pH value over time in the nutrient solution 1 of ppm.
The mass concentration that Figure 13 is illustrated in sodium bicarbonate respectively is adjusted into 0.28 * 10 3Ppm, 0.56 * 10 3Ppm, 1.0 * 10 3Ppm, 2.0 * 10 3Cell concn over time in the nutrient solution 1 of ppm.
The mass concentration that Figure 14 is illustrated in sodium bicarbonate respectively is adjusted into 0.28 * 10 3Ppm, 0.56 * 10 3Ppm, 1.0 * 10 3Ppm, 2.0 * 10 3The pH value over time in the nutrient solution 1 of ppm.
The mass concentration that Figure 15 is illustrated in sodium bicarbonate respectively is adjusted into 1.0 * 10 3Ppm, 7.0 * 10 3Cell concn over time in the nutrient solution 1 of ppm.
The mass concentration that Figure 16 is illustrated in sodium bicarbonate respectively is adjusted into 1.0 * 10 3Ppm, 7.0 * 10 3The pH value over time in the nutrient solution 1 of ppm.
Figure 17 illustrate respectively embodiment 1 to 4 nutrient solution 1 to 4 cell concn over time.
Figure 18 illustrate respectively embodiment 1 to 4 nutrient solution 1 to 4 the pH value over time.
Figure 19 illustrate respectively reappear nutrient solution 1 to 4 in the test cell concn over time.
Figure 20 illustrate respectively reappear nutrient solution 1 to 4 in the test the pH value over time.
Figure 21 illustrate respectively embodiment 5 to 7 nutrient solution 2,4,5 cell concn over time.
Figure 22 illustrate respectively embodiment 5 to 7 nutrient solution 2,4,5 the pH value over time.
The cell concn that Figure 23 illustrates nutrient solution 2,4,5 in the culture experiment respectively again over time.
The pH value that Figure 24 illustrates nutrient solution 2,4,5 in the culture experiment respectively again over time.
Figure 25 is the schema of the semi-batch cultured continuously of use among the embodiment 8.
The cell concn that Figure 26 illustrates nutrient solution 4,6 among the embodiment 8 to 9 respectively over time.
The pH value that Figure 27 illustrates nutrient solution 4,6 among the embodiment 8 to 9 respectively over time.
Figure 28 illustrates among the embodiment 8 to 9 per 1.4 * 10 4The phosphorus aggregate consumption of individual Chaetoceros cell correspondence and the ratio of nitrogen aggregate consumption, and the ratio of silicon aggregate consumption and nitrogen aggregate consumption.
The cell concn that Figure 29 illustrates nutrient solution 4 among the embodiment 10 over time.
The pH value that Figure 30 illustrates nutrient solution 4 among the embodiment 10 over time.
Figure 31 illustrates among the embodiment 10 per 1.4 * 10 4The phosphorus aggregate consumption of individual Chaetoceros cell correspondence and the ratio of nitrogen aggregate consumption, and the ratio of silicon aggregate consumption and nitrogen aggregate consumption.
Figure 32 is the schema of the variation of semi-batch cultured continuously.
Figure 33 is the schema of other variation of semi-batch cultured continuously.
The one-tenth that Figure 34 illustrates the nutrient solution 10,11 that uses in the present embodiment respectively is grouped into.
Figure 35 illustrate respectively embodiment 11 and comparative example nutrient solution 10,11 cell concn over time.
Figure 36 illustrate respectively embodiment 11 and comparative example nutrient solution 10,11 the pH value over time.
Embodiment
In the present embodiment, carried out 5 tests and parsing at first as pretesting, be used for condition stable and that diatom is cultivated on high density ground with searching.Then, based on these conditions, seek method stable and that diatom is cultivated on high density ground.
(pretesting)
1) record is seeded in the variation (proliferative amount) and the relation that constitutes this nutrient solution composition consumption of the Chaetoceros cultivation amount in the liquid (nutrient solution).
Preparation has nutrient solution composition shown in Figure 1 and mutually different 11 samples of composition ratio of components.Each sample put into have specified volume (for example about 65 * 10 -3m 3) outdoor use (natural light with) hermetic type reactor in.The nutrient solution capacity of each sample is certain.Then, the Chaetoceros with the regulation cell count is seeded in each reactor.Next, through after the specified time, measure cell concn variation (culturing cell concentration-incipient cell concentration, the unit: cell/m of each sample 3), nitrogen consumption, phosphorus consumption and silicon consumption.Nitrogen (N), phosphorus (P) and silicon (Si) are the important composition elements during Chaetoceros is cultivated.
With the water-soluble artificial seawater that becomes of marine organisms shown in Figure 1 (arineTech of M Co., Ltd. system).Need to prove to be dissolved in the composition that becomes artificial seawater in the water, also can use marine organisms composition in addition.And be not defined as artificial seawater, also can implement the natural sea-water of sterilising treatment for using autoclave, filtration sterilization or medicine etc., or unsterilized natural sea-water.
Because the capacity of each nutrient solution is certain, so the nitrogen consumption is represented with the mass concentration (nitrogen concentration of initial-stage culture liquid-residual nitrogen concentration) of consumption.The mass concentration unit of nitrogen is (ppm).
Because the capacity of each nutrient solution is certain, so the phosphorus consumption is represented with the mass concentration (the phosphorus mass concentration of initial-stage culture liquid-residual phosphorus mass concentration) of consumption.The mass concentration unit of phosphorus is (ppm).
Because the capacity of each nutrient solution is certain, so the silicon consumption is represented with the mass concentration (the siliceous amount concentration of initial-stage culture liquid-residual siliceous amount concentration) of consumption.The mass concentration unit of silicon is (ppm).
Fig. 2 illustrates each nutrient solution cell concn variation (cell/m in 11 samples 3), nitrogen consumption, phosphorus consumption and silicon consumption.
Based on this measurement result, the relation that relation that the relation that nitrogen consumption and Chaetoceros cell concn change, phosphorus consumption and Chaetoceros cell concn change and silicon consumption and Chaetoceros cell concn change is respectively shown in Fig. 3 A, Fig. 3 B and Fig. 3 C.
The relation of each consumption of nitrogen, phosphorus and silicon and cell concn variable quantity (proliferative amount) is shown in two long and short dash line of Fig. 3 A, Fig. 3 B and Fig. 3 C, and composition concerns.
2) value of P/N (mass concentration ratio of phosphorus and nitrogen) and the value of Si/N (mass concentration ratio of silicon and nitrogen) in the record nutrient solution composition.
In the document 1 and 2 that discloses existing cultural method, put down in writing each mass concentration of nitrogen, phosphorus and silicon in the Chaetoceros nutrient solution.The mass concentration of the mass concentration of the mass concentration of nitrogen, phosphorus amount, silicon amount is respectively 13.8 (ppm), 1.4 (ppm), 0.15 (ppm) in the nutrient solution of document 1.The mass concentration of the mass concentration of the mass concentration of nitrogen, phosphorus amount, silicon amount is respectively 102 (ppm), 1.4 (ppm), 8.0 (ppm) in the nutrient solution of document 2.
P/N, Si/N are respectively 0.10,0.011 in the nutrient solution of document 1.P/N, Si/N are respectively 0.014,0.078 in the nutrient solution of document 2.
Shown in Fig. 3 A, 3B, 3C, because each consumption and the cell concn variable quantity of nitrogen, phosphorus and silicon are proportional, if therefore infer P/N and Si/N be set at the value of calculating greater than by document 1 and document 2 respectively, promptly be at least more than 0.1 or 0.1, then compare, can increase the proliferative amount of Chaetoceros with existing cultural method.
In addition, shown in the dotted line of Fig. 3 A, 3B, 3C, can infer that the cell concn in order to make Chaetoceros is about 6.0 * 10 13Cell/m 3Or 6.0 * 10 13Cell/m 3More than, preferably use following synthetic nutrient solution, that is, make the mass concentration of nitrogen be about 160ppm or 160ppm mass concentration above, phosphorus is about 20ppm or 20ppm mass concentration above and silicon is about 60ppm or more than the 60ppm.
In order to verify above-mentioned 2 suppositions, modulation has nutrient solution 1～nutrient solution 6 that one-tenth shown in Figure 4 is grouped into.The one-tenth that the composition that need to prove nutrient solution 7 consists of document 1 described nutrient solution is grouped into, and the one-tenth that the composition of nutrient solution 8 consists of document 2 described nutrient solutions is grouped into.The nitrogen concentration of each nutrient solution, phosphorus mass concentration, siliceous amount concentration, P/N and Si/N are respectively as shown in Figure 5.
The phosphorus mass concentration of nutrient solution 1 and siliceous amount concentration are less than above-mentioned guess value.In nutrient solution 1, nitrogen concentration, phosphorus mass concentration, siliceous amount concentration are respectively 166 (ppm), 10 (ppm), 30 (ppm).P/N, Si/N are respectively 0.06,0.18.
Nutrient solution 2 be modulated to 2 times of one-tenth of phosphorus mass concentration to nutrient solution 1 be grouped into (phosphorus mass concentration=20 (ppm), P/N=0.12).
Nutrient solution 3 be modulated to 2 times of one-tenth of siliceous amount concentration to nutrient solution 1 be grouped into (siliceous amount concentration=60 (ppm), P/N=0.36).
Nutrient solution 4 be modulated to phosphorus mass concentration and siliceous amount concentration respectively 2 times of one-tenth to nutrient solution 1 be grouped into (mass concentration of phosphorus amount=20 (ppm), P/N=0.12, the mass concentration of silicon amount=60 (ppm), P/N=0.36).
Nutrient solution 5 be modulated to 3 times of one-tenth of siliceous amount concentration to nutrient solution 2 be grouped into (mass concentration of silicon amount=90 (ppm), P/N=0.54).
Nutrient solution 6 be modulated to 4 times of one-tenth of siliceous amount concentration to nutrient solution 2 be grouped into (mass concentration of silicon amount=120 (ppm), P/N=0.72).
Phosphorus mass concentration in the nutrient solution 2～6, siliceous amount concentration, P/N, Si/N all are higher than the phosphorus mass concentration in the nutrient solution 7,8 with existing moiety, siliceous amount concentration, P/N, Si/N.
3) relation of pH value and amount of hydrolysis in the nutrient solution that record phosphorus mass concentration and siliceous amount concentration are high.
Adjustment is as the sodium bicarbonate (NaHCO of one of nutrient solution composition 3) mass concentration, the pH of nutrient solution 4 is changed between 7.0～9.0.At this moment, the amount of hydrolysis in the mensuration nutrient solution 4 is (with respect to 1m 3Solution, sedimentary hydrolyzate quality (kg)).Need to prove that the limit will stirring be supplied to nutrient solution 4 with air as nutrient solution as the air neccessary composition that earns a bare living in the nutrient solution, that mixed 3% carbonic acid gas, aeration-agitation is carried out to nutrient solution 4 in the limit.
Fig. 6 illustrates the graphic representation of this measurement result.The solid line of Fig. 6 and dotted line illustrate the amount of hydrolysis that obtains different tests day respectively to be changed, and obtains roughly the same result.
Thus measurement result as can be known, if the pH of nutrient solution 4 surpasses 8.5, then amount of hydrolysis changes violently, amount of hydrolysis (precipitation capacity) increases.If the pH of nutrient solution 4 surpasses 8.5, then contain Chaetoceros, so Chaetoceros can't be bred owing in the throw out.
4) determine the pH higher limit of nutrient solution.Use is as the sodium bicarbonate of one of nutrient solution composition and the pH of the air adjustment nutrient solution that has mixed 3% carbonic acid gas.Usually, if the amount of sodium bicarbonate increases, then the pH value of solution increases, and in addition, if concentration of carbon dioxide increases, then the pH of solution reduces.Need to prove that sodium bicarbonate and carbonic acid gas are the photosynthetic carbon sources of Chaetoceros, and have the effect of nutrient solution pH regulator agent.
With nutrient solution 1 (110 * 10 -3M3) put into and place outdoor hermetic type reactor, at about about 25 ℃, (the photon amount is about 1200 μ mol/m with the sunlight of nature on the limit with temperature maintenance on the limit 2The light intensity of/s) irradiation nutrient solution 1.Under this state, carry out following 2 mensuration, i.e. i) with having mixed the air aeration-agitation nutrient solution 1 of 3% carbonic acid gas, measure respectively that the Chaetoceros cell concn reaches pH over time over time in the nutrient solution 1; Ii) add sodium bicarbonate (7.0 * 10 3Ppm), and nutrient solution 1 is carried out aeration-agitation, measure respectively that the Chaetoceros cell concn reaches pH over time over time in the nutrient solution 1 with air.
Fig. 8 is the graphic representation that this measurement result is shown.The solid line of Fig. 8 is represented the 1st measurement result, does not promptly add sodium bicarbonate, carries out under the situation of aeration-agitation with the air that has mixed 3% carbonic acid gas, and the pH time of nutrient solution 1 changes; The dotted line of Fig. 8 is represented the 2nd measurement result, promptly adds sodium bicarbonate, only carries out under the situation of aeration-agitation with air, and the pH time of nutrient solution 1 changes.
By measurement result as can be known, nutrient solution 1 is carried out aeration-agitation, and add under the situation of sodium bicarbonate,, precipitation then occurs if the pH of nutrient solution 1 surpasses 8.5 with air.At this moment, Chaetoceros and throw out together precipitate.In addition, carry out aeration-agitation with the air that has mixed 3% carbonic acid gas, and do not add under the situation of sodium bicarbonate, the pH of nutrient solution 1 does not take place to reduce gradually.
By 3) and 4) measurement result as can be known, and the situation that nutrient solution carries out aeration-agitation is compared with the air that has mixed 3% carbonic acid gas, adding sodium bicarbonate in nutrient solution can be set at pH below 8.4 or 8.4.Its result is for preventing the co-precipitation of sedimentary appearance and Chaetoceros respectively.
The composition that need to prove nutrient solution 1 is compared with the composition of nutrient solution 2～6, and except P/N and Si/N, other compositions are identical.So, because the solubleness of nutrient solution 1 and the solubleness of other nutrient solutions are roughly the same, therefore infer for other nutrient solutions, in nutrient solution, add under the state of sodium bicarbonate, by nutrient solution being carried out aeration-agitation, also can prevent the co-precipitation of sedimentary appearance and Chaetoceros respectively with the air that has mixed carbonic acid gas.
5) lower value of record nutrient solution pH and the suitable addition scope of sodium bicarbonate.
Adjustment is prepared following 8 kinds of nutrient solutions 1 as the mass concentration of the sodium bicarbonate of one of nutrient solution composition, with the air that has mixed 3% carbonic acid gas each nutrient solution 1 is carried out aeration-agitation: [1] 0.02 * 10 3Ppm, [2] 0.07 * 10 3Ppm, [3] 0.14 * 10 3Ppm, [4] 0.28 * 10 3Ppm, [5] 0.56 * 10 3Ppm, [6] 1.0 * 10 3Ppm, [7] 2.0 * 10 3Ppm, [8] 7.0 * 10 3Ppm.Each nutrient solution 1 that as above is modulated into is carried out the Chaetoceros culture experiment, measure the time variation of cell concn and the time variation of pH.Need to prove in order to ensure circulation ratio, change the date, carry out 4 above-mentioned tests.
Fig. 9 and 10 is the 1st measurement result.In the 1st culture experiment, use nutrient solution 1:[1 with following 3 kinds of sodium bicarbonate mass concentrations] 0.02 * 10 3Ppm, [3] 0.14 * 10 3Ppm, [6] 1.0 * 10 3Ppm.X1 shown in Figure 9, X2, X3 are illustrated respectively in [1] 0.02 * 10 3Ppm, [3] 0.14 * 10 3Ppm, [6] 1.0 * 10 3The time of the Chaetoceros cell concn of cultivating in the nutrient solution 1 of ppm changes.XA1 shown in Figure 10, XA2, XA3 represent [1] 0.02 * 10 respectively 3Ppm, [3] 0.14 * 10 3Ppm, [6] 1.0 * 10 3The pH of the nutrient solution 1 of ppm over time.
Figure 11 and Figure 12 are the 2nd measurement result.In the 2nd culture experiment, use nutrient solution 1:[2 with following 4 kinds of sodium bicarbonate mass concentrations] 0.07 * 10 3Ppm, [3] 0.14 * 10 3Ppm, [4] 0.28 * 10 3Ppm, [6] 1.0 * 10 3Ppm.Y1 shown in Figure 11, Y2, Y3, Y4 are illustrated respectively in [2] 0.07 * 10 3Ppm, [3] 0.14 * 10 3Ppm, [4] 0.28 * 10 3Ppm, [6] 1.0 * 10 3The time of the Chaetoceros cell concn of cultivating in the nutrient solution 1 of ppm changes.YA1 shown in Figure 12, YA2, YA3, YA4 represent [2] 0.07 * 10 respectively 3Ppm, [3] 0.14 * 10 3Ppm, [4] 0.28 * 10 3Ppm, [6] 1.0 * 10 3The pH of the nutrient solution 1 of ppm over time.
Figure 13 and Figure 14 are the 3rd measurement result.In the 3rd culture experiment, use nutrient solution 1:[4 with following 4 kinds of sodium bicarbonate mass concentrations] 0.28 * 10 3Ppm, [5] 0.56 * 10 3Ppm, [6] 1.0 * 10 3Ppm, [7] 2.0 * 10 3Ppm.Z1 shown in Figure 13, Z2, Z3, Z4 are illustrated respectively in [4] 0.28 * 10 3Ppm, [5] 0.56 * 10 3Ppm, [6] 1.0 * 10 3Ppm, [7] 2.0 * 10 3The Chaetoceros cell concn of cultivating in the nutrient solution 1 of ppm over time.ZA1 shown in Figure 14, ZA2, ZA3, ZA4 represent [4] 0.28 * 10 respectively 3Ppm, [5] 0.56 * 10 3Ppm, [6] 1.0 * 10 3Ppm, [7] 2.0 * 10 3The pH of the nutrient solution 1 of ppm over time.
Figure 15 and Figure 16 are the 4th measurement result.In the 4th culture experiment, use nutrient solution 1:[6 with following 2 kinds of sodium bicarbonate mass concentrations] 1.0 * 10 3Ppm, [8] 7.0 * 10 3Ppm.W1 shown in Figure 15, W2 are illustrated respectively in [6] 1.0 * 10 3Ppm, [8] 7.0 * 10 3The Chaetoceros cell concn of cultivating in the nutrient solution 1 of ppm over time.WA1 shown in Figure 16, WA2 represent [6] 1.0 * 10 respectively 3Ppm, [8] 7.0 * 10 3The pH of the nutrient solution 1 of ppm over time.
Measurement result is as can be known thus:
(1) 0.02 * 10 3In the nutrient solution 1 of ppm, the pH value reduces after test at once, reduce to 6.4 after about 25 hours, and cell concn does not reach 3 * 10 13Cell/m 3
(2) 0.07 * 10 3In the nutrient solution 1 of ppm, the pH value maintains in the scope more than about 6.4 or 6.4, below about 7.0 or 7.0, and cell concn reaches 3 * 10 13Cell/m 3
(3) 0.14 * 10 3In the nutrient solution 1 of ppm, the pH value maintains in the scope more than about 6.4 or 6.4, below about 7.4 or 7.4, and cell concn surpasses 4 * 10 13Cell/m 3
(4) 0.28 * 10 3In the nutrient solution 1 of ppm, the pH value maintains in the scope more than about 6.4 or 6.4, below about 7.6 or 7.6, and cell concn surpasses 4 * 10 13Cell/m 3
(5) 0.56 * 10 3In the nutrient solution 1 of ppm, the pH value maintains in the scope more than about 7.0 or 7.0, below about 7.6 or 7.6, and cell concn surpasses 4 * 10 13Cell/m 3
(6) 1.0 * 10 3In the nutrient solution 1 of ppm, the pH value maintains in the scope more than about 6.4 or 6.4, below about 7.9 or 7.9, and cell concn surpasses 6 * 10 13Cell/m 3
(7) 2.0 * 10 3In the nutrient solution 1 of ppm, the pH value maintains in the scope more than about 7.0 or 7.0, below about 8.1 or 8.1, and cell concn surpasses 5 * 10 13Cell/m 3
(8) 7.0 * 10 3In the nutrient solution 1 of ppm, the pH value maintains in the scope more than about 6.7 or 6.7, below about 8.1 or 8.1, and cell concn surpasses 3 * 10 13Cell/m 3
Therefore, have greater than 0.02 * 10 being adjusted to 3Behind the nutrient solution 1 of the sodium bicarbonate mass concentration of ppm, with the air that has mixed 3% carbonic acid gas nutrient solution 1 is carried out aeration-agitation, the pH of nutrient solution is set at more than 6.4 or 6.4, makes the higher limit (3 * 10 of the cell concn of Chaetoceros thus above the Chaetoceros cell concn of existing cultural method 13Cell/m 3).
The composition that need to prove nutrient solution 1 is compared with the composition of nutrient solution 2～6, and except P/N and/or Si/N, other compositions are identical.Therefore, for other nutrient solutions, similarly be adjusted into and have 0.02 * 10 3Ppm or 0.02 * 10 3Behind the nutrient solution 1 of the sodium bicarbonate mass concentration that ppm is above, with the air that has mixed 3% carbonic acid gas nutrient solution 1 is carried out aeration-agitation, the pH of nutrient solution 1 is set at more than 6.4 or 6.4, makes the higher limit (3 * 10 of the cell concn of Chaetoceros thus above the Chaetoceros cell concn of existing cultural method 13Cell/m 3).
By above-mentioned 4 measurement results as can be known, when using nutrient solution 1～6 to cultivate Chaetoceros, be set in the scope more than 6.4 or 6.4, below 8.4 or 8.4, can stably cultivate Chaetoceros by pH with nutrient solution.In order to satisfy this condition, for example modulation has and is higher than 0.02 * 10 3The nutrient solution of the sodium bicarbonate mass concentration of ppm carries out aeration-agitation with the air that has mixed 3% carbonic acid gas to nutrient solution.
Be set at more than 6.4 or 6.4 by pH, can surpass the higher limit of the Chaetoceros cell concn of existing cultural method nutrient solution 1～6.In addition, be set at below 8.4 or 8.4, can suppress to result from that pH increases and the generation of the Chaetoceros co-precipitation of the hydrolyzate that generates prevents that the propagation of Chaetoceros from reducing by pH with nutrient solution 1～6.
Next, based on this condition, seek method stable and that diatom is cultivated on high density ground by embodiment 1～11.
(embodiment 1)
With nutrient solution 1 (1.5 * 10 -3m 3) join laboratory shown in Figure 7 with flat culturing bottle 11 (capacity 1.5 * 10 -3m 3) in, interpolation sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 in nutrient solution 1 3Ppm.Then, the Chaetoceros of inoculation specified amount in nutrient solution 1.Then, the temperature maintenance of nutrient solution at about 25 ℃～about 35 ℃, and is utilized luminescent lamp etc., make to have the about 200 μ mol/m of photon amount 2The rayed nutrient solution 1 of/s intensity under this state, via the Glass tubing 13 that the bolt 12 with bottle 11 is communicated with, is sent into the air that has mixed 3% carbonic acid gas in the nutrient solution 1, and aeration-agitation is carried out to nutrient solution 1 in the limit, and the culture experiment of Chaetoceros is carried out on the limit.Measure the time variation of Chaetoceros cell concn in the nutrient solution 1 and the time variation of pH respectively.
(embodiment 2)
With nutrient solution 2 (1.5 * 10 -3m 3) joining the laboratory with in the flat culturing bottle 11, interpolation sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, in nutrient solution 2, inoculate specified amount (with embodiment 1 with amount) Chaetoceros, the condition roughly the same with embodiment 1 (temperature of nutrient solution: about 25 ℃～about 35 ℃, photon amount: about 200 μ mol/m 2/ s carries out aeration-agitation by the air that has mixed 3% carbonic acid gas) under, carry out the culture experiment of Chaetoceros.Measure the time variation of Chaetoceros cell concn in the nutrient solution 2 and the time variation of pH respectively.
(embodiment 3)
With nutrient solution 3 (1.5 * 10 -3m 3) join the laboratory with flat culturing bottle 11, adding sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, in nutrient solution 3, inoculate specified amount (with embodiment 1 with amount) Chaetoceros, the condition roughly the same with embodiment 1 (culture-liquid temp: about 25 ℃～about 35 ℃, photon amount: about 200 μ mol/m 2/ s carries out aeration-agitation by the air that has mixed 3% carbonic acid gas) under, carry out the culture experiment of Chaetoceros.Measure the time variation of Chaetoceros cell concn in the nutrient solution 3 and the time variation of pH respectively.
(embodiment 4)
With nutrient solution 4 (1.5 * 10 -3m 3) joining the laboratory with in the flat culturing bottle 11, interpolation sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, in nutrient solution 4, inoculate specified amount (with embodiment 1 with amount) Chaetoceros, the condition roughly the same with embodiment 1 (culture-liquid temp: about 25 ℃～about 35 ℃, photon amount: about 200 μ mol/m 2/ s carries out aeration-agitation by the air that has mixed 3% carbonic acid gas) under, carry out the culture experiment of Chaetoceros.Measure the time variation of Chaetoceros cell concn in the nutrient solution 4 and the time variation of pH respectively.
Figure 17 illustrates the graphic representation that the Chaetoceros cell concn time changes among the embodiment 1～4.C1 shown in Figure 17, C2, C3, C4 represent that respectively the time of the Chaetoceros cell concn of cultivation in the nutrient solution 1,2,3,4 changes.In addition, A shown in Figure 17 is illustrated in supply in each nutrient solution and topdresses time of composition (nitrogen that contains in the nutrient solution, phosphorus, silicon, vitamins, mineral substance class).
Figure 18 illustrates the graphic representation that the pH time changes among the embodiment 1～4.D1 shown in Figure 180, D2, D3, D4 represent that respectively the time of pH in the nutrient solution 1,2,3,4 changes.
Test-results is as can be known thus:
(P/N=0.06, Si/N=0.18), the silicon amount is set at 2 times nutrient solution 3, and (P/N=0.06 Si/N=0.36) obtains than Chaetoceros cell concn in the nutrient solution 1 (about 6.0 * 10 with respect to nutrient solution 1 13Cell/m 3) high cell concn (about 8.0 * 10 13Cell/m 3).
(P/N=0.06, Si/N=0.18), phosphorus amount and silicon amount are set at 2 times nutrient solution 4 respectively, and (P/N=0.12 Si/N=0.36) obtains than Chaetoceros cell concn in the nutrient solution 1 (about 6.0 * 10 with respect to nutrient solution 1 13Cell/m 3) high cell concn (about 7.0 * 10 13Cell/m 3).
Each pH of nutrient solution 1～4 maintains the scope more than 6.4 or 6.4, below 8.5 or 8.5, more specifically, maintains in the scope more than about 7.5 or 7.5, below about 8.5 or 8.5.
Need to prove that the supply that has mixed the air of carbonic acid gas in the culture experiment may be abundant inadequately for the cell concn of nutrient solution 4 among the embodiment 4 is measured, with practical situation may be by many differences.
In order to study the circulation ratio of embodiment 1～4 measurement result, with the same condition of embodiment 1～4 under, the not supply composition that topdresses, the time of measuring Chaetoceros cell concn in the nutrient solution 1～4 more respectively changes and the time of pH changes.
Figure 19 illustrates to reappear the graphic representation that the Chaetoceros cell concn time changes in the nutrient solution 1～4 that uses in the test.E1 shown in Figure 19, E2, E3, E4 represent that respectively the time of the Chaetoceros cell concn of cultivation in the nutrient solution 1,2,3,4 changes.
Figure 20 illustrates to reappear the graphic representation that the pH time changes in the nutrient solution 1～4 that uses in the test.F1 shown in Figure 20, F2, F3, F4 represent that respectively the time of pH in the nutrient solution 1,2,3,4 changes.
Test-results is as can be known thus:
(P/N=0.06, Si/N=0.18), the phosphorus amount is set at 2 times nutrient solution 3, and (P/N=0.12 Si/N=0.18) obtains than the high cell concn of Chaetoceros cell concn in the nutrient solution 1 with respect to nutrient solution 1.
(P/N=0.06, Si/N=0.18), the silicon amount is set at 2 times nutrient solution 3, and (P/N=0.06 Si/N=0.36) obtains than the high cell concn of Chaetoceros cell concn in the nutrient solution 1 with respect to nutrient solution 1.
(P/N=0.06, Si/N=0.18), phosphorus amount and silicon amount are set at 2 times nutrient solution 4 respectively, and (P/N=0.12 Si/N=0.36) obtains than the high cell concn of Chaetoceros cell concn in the nutrient solution 1 with respect to nutrient solution 1.
Nutrient solution 4 obtains than the high cell concn of Chaetoceros cell concn in the nutrient solution 2,3.
Each pH of nutrient solution 1～4 maintains the scope more than 6.4 or 6.4, below 8.5 or 8.5, more specifically, maintains in the scope more than about 7.4 or 7.4, below about 8.0 or 8.0.
Can infer if topdress by the result of the foregoing description 1～4 and to contain phosphorus amount more than 2 times or 2 times and/or the nutrient solution of silicon amount more than 2 times or 2 times that cultivate Chaetoceros, then the cell concn of Chaetoceros is about 6.0 * 10 with respect to nutrient solution 1 13Cell/m 3Or 6.0 * 10 13Cell/m 3More than.
In order to confirm above-mentioned supposition, topdress with respect to nutrient solution 1 contain 2 times of phosphorus amounts nutrient solution 2, with respect to nutrient solution 1 contain 2 times of phosphorus amounts and 2 times of silicon amounts nutrient solution 4, contain the nutrient solution 5 of 2 times of phosphorus amounts and 3 times of silicon amounts with respect to nutrient solution 1, carry out the cultivation of Chaetoceros respectively.
(embodiment 5)
Similarly to Example 2, with nutrient solution 2 (1.5 * 10 -3m 3) join in the flat culturing bottle 11, interpolation sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, in nutrient solution 2, inoculate specified amount (with embodiment 2 with amount) Chaetoceros, the condition roughly the same with embodiment 2 (culture-liquid temp: about 25 ℃～about 35 ℃, photon amount: about 200 μ mol/m 2/ s carries out aeration-agitation by the air that has mixed 3% carbonic acid gas) under, carry out the culture experiment of Chaetoceros.Measure the time variation of Chaetoceros cell concn in the nutrient solution 2 and the time variation of pH respectively.
(embodiment 6)
Similarly to Example 4, with nutrient solution 4 (1.5 * 10 -3m 3) join in the flat culturing bottle 11, interpolation sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, in nutrient solution 4, inoculate specified amount (with embodiment 5 with amount) Chaetoceros, the condition roughly the same with embodiment 5 (culture-liquid temp: about 25 ℃～about 35 ℃, photon amount: about 200 μ mol/m 2/ s carries out aeration-agitation by the air that has mixed 3% carbonic acid gas) under, carry out the culture experiment of Chaetoceros.Measure the time variation of Chaetoceros cell concn in the nutrient solution 4 and the time variation of pH respectively.
(embodiment 7)
With nutrient solution 5 (1.5 * 10 -3m 3) joining flat culturing bottle 11, interpolation sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, in nutrient solution 5, inoculate specified amount (with embodiment 5 with amount) Chaetoceros, the condition roughly the same with embodiment 5 (culture-liquid temp: about 25 ℃～about 35 ℃, photon amount: about 200 μ mol/m 2/ s carries out aeration-agitation by the air that has mixed 3% carbonic acid gas) under, carry out the culture experiment of Chaetoceros.Measure the time variation of Chaetoceros cell concn in the nutrient solution 5 and the time variation of pH respectively.
Figure 21 illustrates the graphic representation that the Chaetoceros cell concn time changes among the embodiment 5～7.G1 shown in Figure 21, G2, G3, G4 represent that respectively the time of the Chaetoceros cell concn of cultivation in the nutrient solution 2,4,5 changes.In addition, A shown in Figure 21 is illustrated in supply in each nutrient solution and topdresses time of composition (nitrogen that contains in the nutrient solution, phosphorus, silicon, vitamins, mineral substance class).
Figure 22 illustrates the graphic representation that the pH time changes among the embodiment 5～7.H1 shown in Figure 22, H2, H3 represent that respectively the time of pH in the nutrient solution 2,4,5 changes.In addition, H4 shown in Figure 22 represents the time variation of room temperature in the laboratory, and H5 represents that the time of culture-liquid temp changes.
Measurement result is as can be known thus:
(P/N=0.12, Si/N=0.18), the silicon amount is set at 2 times nutrient solution 4, and (P/N=0.12 Si/N=0.36) obtains cell concn (about 9.0 * 10 than the Chaetoceros cell concn is high in the nutrient solution 2 with respect to nutrient solution 2 13Cell/m 3).
(P/N=0.12, Si/N=0.18), the silicon amount is set at 3 times nutrient solution 5, and (P/N=0.12 Si/N=0.54) obtains cell concn (about 9.0 * 10 than the Chaetoceros cell concn is high in the nutrient solution 2 with respect to nutrient solution 2 13Cell/m 3).
In the nutrient solution 4 in the cell concn of Chaetoceros and the nutrient solution 5 cell concn of Chaetoceros do not have than big-difference.
Each pH of nutrient solution 2,4,5 maintains the scope more than 6.4 or 6.4, below 8.5 or 8.5, more specifically, maintains in the scope more than about 7.4 or 7.4, below about 8.2 or 8.2.
Can infer that by above-mentioned test-results then the cell concn of Chaetoceros is about 8.0 * 10 if use the nutrient solution cultivation Chaetoceros of the silicon amount more than 2 times or 2 times that contains with respect to nutrient solution 2 13Cell/m 3Or 8.0 * 10 13Cell/m 3More than.
In order to confirm above-mentioned supposition, except nutrient solution 2,4,5, use nutrient solution 6, under the condition identical, carry out culture experiment once more with embodiment 5, the time of measuring Chaetoceros cell concn in the nutrient solution 2,4,5,6 respectively changes and the time of pH changes.
Figure 23 illustrates the graphic representation that the Chaetoceros cell concn time changes in the culture experiment again.I1 shown in Figure 23, I2, I3, I4 represent that respectively the time of the Chaetoceros cell concn of cultivation in the nutrient solution 2,4,5,6 changes.
Figure 24 illustrates the graphic representation that the pH time changes in the culture experiment again.J1 shown in Figure 24, J2, J3, J4 represent the pH of nutrient solution 2,4,5,6 respectively.In addition, J5 shown in Figure 24 represents the time variation of room temperature in the laboratory, and J6 represents that the time of culture-liquid temp changes.
Test-results is as can be known thus:
(P/N=0.12 Si/N=0.18), is set at 2 times nutrient solution 4 (Si/N=0.36) with the silicon amount and obtains cell concn (about 1.0 * 10 than the Chaetoceros cell concn is high in the nutrient solution 2 with respect to nutrient solution 2 14Cell/m 3).
(P/N=0.12 Si/N=0.18), is set at 3 times nutrient solution 5 (Si/N=0.54) with the silicon amount and obtains cell concn (about 9.0 * 10 than the Chaetoceros cell concn is high in the nutrient solution 2 with respect to nutrient solution 2 13Cell/m 3).
(P/N=0.12 Si/N=0.18), is set at 4 times nutrient solution 6 (Si/N=0.72) with the silicon amount and obtains cell concn (about 1.0 * 10 than the Chaetoceros cell concn is high in the nutrient solution 2 with respect to nutrient solution 2 14Cell/m 3).
In the nutrient solution 4 in Chaetoceros cell concn and the nutrient solution 5,6 the Chaetoceros cell concn do not have than big-difference.
Each pH of nutrient solution 2,4,5,6 maintains the scope more than 6.4 or 6.4, below 8.5 or 8.5, more specifically, maintains in the scope more than about 7.3 or 7.3, below about 8.1 or 8.1.
By the test-results of the foregoing description 5～7 as can be known, cultivate Chaetoceros by using a kind of nutrient solution in the nutrient solution 4～6, the cell concn of Chaetoceros increases to and is about 8.0 * 10 13Cell/m 3Or 8.0 * 10 13Cell/m 3More than.In other words, be more than 0.12 or 0.12, reach the nutrient solution cultivation Chaetoceros that Si/N is a value more than 0.36 or 0.36 by using P/N, the cell concn of Chaetoceros increases to and is about 8.0 * 10 13Cell/m 3Or 8.0 * 10 13Cell/m 3More than.
Next, in order to study when the cultured continuously, whether culture condition (P/N 〉=0.12, and Si/N 〉=0.36) is all set up indoor and outdoor, uses nutrient solution 4 and nutrient solution 6, implements the test based on embodiment 8～10.
(embodiment 8)
The nutrient solution 4 (1.5 * 10 that will have nitrogen (166ppm), phosphorus (20ppm), silicon (60ppm) -3m 3) join the laboratory with flat culturing bottle 11, adding sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, the Chaetoceros of inoculation specified amount in nutrient solution 4, the condition roughly the same with embodiment 1 (culture-liquid temp: about 25 ℃～about 35 ℃, photon amount: about 200 μ mol/m 2/ s carries out aeration-agitation by the air that has mixed 3% carbonic acid gas) under, carry out the culture experiment of Chaetoceros.Measure the time variation of Chaetoceros cell concn in the nutrient solution 4 and the time variation of pH respectively.
In the mensuration process, if the cell concn of Chaetoceros reaches 6.0 * 10 13Cell/m 3Or 6.0 * 10 13Cell/m 3More than, then from nutrient solution 4, take out the solution that accounts for total amount 2/3.Then, prepare taking-up amount from nutrient solution 4 dissolving halobiontic artificial seawater.By dissolve the composition that topdresses that contains nitrogen, phosphorus, silicon, vitamins etc. in this artificial seawater, making topdresses uses nutrient solution.This is topdressed to go in the nutrient solution 4 in the flat culturing bottle 11 with the nutrient solution supply.This is called the semi-batch cultured continuously.
In the semi-batch cultured continuously, measure respectively from nutrient solution 4 take out account for the solution of total amount 2/3 after, supply is topdressed with nitrogen amount, phosphorus amount and the silicon amount of the preceding nutrient solution 4 (being designated hereinafter simply as the preceding nutrient solution 4a that topdresses) of nutrient solution in nutrient solution 4.Then, obtain respectively topdress before institute's nitrogen content, phosphorus amount and silicon amount poor in the nutrient solution (initial-stage culture liquid) 4 before institute's nitrogen content, phosphorus amount and silicon amount and the inoculation Chaetoceros among the nutrient solution 4a.Need to prove that nitrogen amount in the initial-stage culture liquid 4, phosphorus amount, phosphorus amount are respectively 166ppm, 20ppm, 60ppm.
Poor based on nitrogen amount, phosphorus amount and silicon amount, the topdressing of nitrogen amount, phosphorus amount and silicon amount that the composition that topdresses has been adjusted in supply among the nutrient solution 4a before topdressing used nutrient solution, so that P/N and the Si/N among the nutrient solution 4a is consistent with P/N (=0.12) and Si/N (=0.36) in the initial-stage culture liquid 4 respectively before topdressing.By this operation, make P/N and Si/N in the nutrient solution 4 before and after supply, remain certain (with reference to Figure 25) respectively.
The semi-batch cultured continuously is repeated on the limit, and the limit is measured the time variation of Chaetoceros cell concn in the nutrient solution 4, the time variation of pH respectively, obtains per 1 * 10 4The nitrogen consumption of individual Chaetoceros cell, phosphorus consumption and silicon consumption.
(embodiment 9)
The nutrient solution 6 (1.5 * 10 that will contain nitrogen (166ppm), phosphorus (20ppm), silicon (120ppm) -3m 3) join the laboratory with flat culturing bottle 11, adding sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, in nutrient solution 6, inoculate specified amount (with embodiment 8 with amount) Chaetoceros, the condition roughly the same with embodiment 8 (culture-liquid temp: about 25 ℃～about 35 ℃, photon amount: about 200 μ mol/m 2/ s carries out aeration-agitation by the air that has mixed 3% carbonic acid gas) under, carry out the culture experiment (semi-batch cultivate cultured continuously test) of Chaetoceros.Measure the time variation of Chaetoceros cell concn in the nutrient solution 6 and the time variation of pH respectively, obtain per 1 * 10 4The nitrogen consumption of individual Chaetoceros cell, phosphorus consumption and silicon consumption.
Figure 26 illustrates the graphic representation that the Chaetoceros cell concn time changes in embodiment 8 and 9.K1 shown in Figure 26, K2 represent the cell concn of the Chaetoceros of cultivation in the nutrient solution 4,6 respectively.A1 shown in Figure 26 is illustrated in each nutrient solution initial supply and topdresses time of composition (nitrogen, phosphorus, silicon, vitamins, mineral substance class).At this moment, because the cell concn of Chaetoceros does not reach 6.0 * 10 13Cell/m 3, therefore from each nutrient solution, do not take out the solution that accounts for total amount 2/3.A2 shown in Figure 26～7 represent that respectively the cell concn of Chaetoceros surpasses 6.0 * 10 13Cell/m 3The time from each nutrient solution, take out the solution account for total amount 2/3, supply was topdressed with the time of nutrient solution in each nutrient solution.
Figure 27 illustrates the graphic representation that the pH time changes in embodiment 8 and 9.L1 shown in Figure 27, L2 represent the pH in the nutrient solution 4,6 respectively.
Obtain the nitrogen consumption of the Chaetoceros of each pipeline time in the nutrient solution 4,6, the nitrogen aggregate consumption of Chaetoceros and the Chaetoceros nitrogen aggregate consumption of per unit cell respectively by following formula.
NCOM A1＝NFIR-NREM A1
NCOM A2＝NFIR+NADD A1-NREM A2
NCOM A3＝NREM A2+NADD A2-NREM A3
NCOM A4＝NREM A3+NADD A3-NREM A4
NCOM A5＝NREM A4+NADD A4-NREM A5
NCOM A6＝NREM A5+NADD A5-NREM A6
NCOM A7＝NREM A6+NADD A6-NREM A7
N ALL＝∑ 1≤i≤7(NCOM Ai/CELL Ai)
Herein, NCOM Ai, NFIR, NREM Ai, NADD Ai, NCOM UNIT, CELL Ai, N ALLRepresent following amount respectively.NCOM AiBe illustrated in the nitrogen amount (ppm) of consuming by Chaetoceros between pipeline time A (i～1)～Ai.NFIR represents the nitrogen amount (ppm) in the initial-stage culture liquid.NREM AiResidual nitrogen amount (ppm) in nutrient solution during expression pipeline time Ai.NADD AiSupply topdresses with institute's nitrogen content (ppm) in the nutrient solution during expression pipeline time Ai.N ALLExpression is by the total nitrogen (ppm) of the Chaetoceros consumption of per unit cell.CELL AiThe unit cell number of Chaetoceros in the expression pipeline time Ai, the unit cell number is obtained by the cell concn of Chaetoceros in the pipeline time Ai.
Obtain the phosphorus aggregate consumption P that consumes by the Chaetoceros of every unit cell in the nutrient solution 4,6 by same calculating formula ALLAnd silicon aggregate consumption Si ALL
Figure 28 represent in embodiment 8 and 9 in the initial-stage culture liquid nitrogenous mass concentration, the mass concentration of phosphorus, the mass concentration of silicon, P/N, Si/N.And Figure 28 represents by among the embodiment 8 per 1.0 * 10 4The phosphorus aggregate consumption that nitrogen aggregate consumption (ppm), phosphorus aggregate consumption (ppm) and the silicon aggregate consumption (ppm) of individual Chaetoceros cell consumption obtained and the ratio (P of nitrogen aggregate consumption ALL/ N ALL) and the ratio (Si of silicon aggregate consumption and nitrogen aggregate consumption ALL/ N ALL).Need to prove that embodiment 9 also illustrates same result.
Measurement result is as can be known thus:
About 6.0 * 10 13Cell/m 3Near cultured continuously Chaetoceros stably.
The pH of nutrient solution 4,6 maintains in the scope more than 6.4 or 6.4, below 8.5 or 8.5, more specifically, maintains in the scope more than 7.5 or 7.5, below 8.5 or 8.5.
P ALL/ N ALLAnd Si ALL/ N ALLFor with the P/N and the proximate value of Si/N of initial-stage culture liquid 4.In addition, P ALL/ N ALLFor with the proximate value of the P/N of initial-stage culture liquid 6, Si ALL/ N ALLBe value less than the Si/N of initial-stage culture liquid 6.Thereby, if being put into the laboratory, nutrient solution 4 or 6 cultivates Chaetoceros with culturing bottle 11, then can cultivate to high density Chaetoceros continuously and stably.And, under the situation of nutrient solution 4, can not add superfluous silicon and cultivate by the suitableeest composition ratio.
(embodiment 10)
The nutrient solution 4 (65 * 10 that will have nitrogen (166ppm), phosphorus (20ppm), silicon (60ppm) -3m 3) put into and be arranged on outdoor closed reactor, adding sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, the Chaetoceros of inoculation specified amount in nutrient solution 4.Next, culture-liquid temp is being maintained about 15 ℃～about 35 ℃ scope, preferably maintaining 25 ℃, and with sunlight (the photon amount: about 0 μ mol/m of nature 2/ s～1200 μ mol/m 2/ s) shine under the condition of nutrient solution 4, the air that has mixed 3% carbonic acid gas to be sent in the reactor, aeration-agitation is carried out to nutrient solution 4 in the limit, and the culture experiment of Chaetoceros is carried out on the limit.The time variation of Chaetoceros cell concn and the time of pH change in the nutrient solution 4 outside the measuring cell respectively.
In the mensuration process, if the cell concn of Chaetoceros reaches about 5.0 * 10 13Cell/m 3Or 5.0 * 10 13Cell/m 3More than, then from nutrient solution 4, take out account for total amount half (33 * 10 -3m 3) solution.Then, prepare taking-up amount from nutrient solution 4 dissolving halobiontic artificial seawater.Contain the compositions that topdress such as nitrogen, phosphorus, silicon, vitamins by dissolving in this artificial seawater, making topdresses uses nutrient solution.This is topdressed to go in the nutrient solution 4 in the reactor with the nutrient solution supply, carry out the semi-batch cultured continuously.
In the semi-batch cultured continuously, measure respectively from nutrient solution 4 take out account for half solution of total amount after, nitrogen amount, phosphorus amount and the silicon amount of nutrient solution 4a before topdressing.Then, obtain respectively topdress before institute's nitrogen content, phosphorus amount and silicon amount poor in institute's nitrogen content, phosphorus amount and silicon amount and the initial-stage culture liquid 4 among the nutrient solution 4a.Need to prove that nitrogen amount in the initial-stage culture liquid 4, phosphorus amount, silicon amount are respectively 166ppm, 20ppm, 60ppm.
Poor based on nitrogen amount, phosphorus amount and silicon amount, the topdressing of nitrogen amount, phosphorus amount and silicon amount that the composition that topdresses has been adjusted in supply among the nutrient solution 4a before topdressing used nutrient solution, so that P/N and the Si/N among the nutrient solution 4a is consistent with P/N (=0.12) and Si/N (=0.36) in the initial-stage culture liquid 4 respectively before topdressing.By this operation, make P/N and Si/N in the nutrient solution 4 before and after supply, remain certain respectively.
The semi-batch cultured continuously is repeated on the limit, and the limit is measured the time variation of Chaetoceros cell concn in the nutrient solution 4, the time variation of pH respectively, obtains per 1 * 10 4The nitrogen consumption of individual Chaetoceros cell, phosphorus consumption and silicon consumption.
Figure 29 illustrates the graphic representation that the Chaetoceros cell concn time changes among the embodiment 10.A11 shown in Figure 29 is illustrated in each nutrient solution initial supply and topdresses time of composition.At this moment, because the cell concn of Chaetoceros does not reach 5.0 * 10 13Cell/m 3Therefore, from nutrient solution 4, do not take out and account for half solution of total amount.A12～A17 shown in Figure 29 represents that respectively the cell concn of Chaetoceros surpasses 5.0 * 10 13Cell/m 3In time, takes out from nutrient solution 4 and accounts for half solution of total amount, and supply was topdressed with the time of nutrient solution in nutrient solution 4.Figure 30 is the graphic representation that the time variation of pH among the embodiment 10 is shown.
Among the embodiment 10, obtain per unit cell in the nutrient solution 4 (for example 1.0 * 10 by calculating formula similarly to Example 8 4Nitrogen aggregate consumption (the N of Chaetoceros consumption cell) ALL), phosphorus aggregate consumption (P ALL) and silicon aggregate consumption (Si ALL).
Figure 31 represent among the embodiment 10 in the initial-stage culture liquid nitrogenous mass concentration, the mass concentration of phosphorus, the mass concentration of silicon, P/N, Si/N.And Figure 31 represents by among the embodiment 10 per 1.0 * 10 4The phosphorus aggregate consumption that nitrogen aggregate consumption (ppm), phosphorus aggregate consumption (ppm) and the silicon aggregate consumption (ppm) of a cell Chaetoceros consumption obtained and the ratio (P of nitrogen aggregate consumption ALL/ N ALL) and the ratio (Si of silicon aggregate consumption and nitrogen aggregate consumption ALL/ N ALL).
Measurement result is as can be known thus:
About 5.0 * 10 13Cell/m 3Near cultured continuously Chaetoceros stably.
The pH of nutrient solution 4 maintains in the scope more than 6.4 or 6.4, below 8.5 or 8.5, more specifically, maintains in the scope more than 7.4 or 7.4, below 8.2 or 8.2.
P ALL/ N ALLAnd Si ALL/ N ALLFor with the P/N and the proximate value of Si/N of initial-stage culture liquid 4.Thereby,, then can cultivate Chaetoceros continuously and stably in high cell concentration ground if the closed reactor that nutrient solution 4 is put into outdoor use is cultivated Chaetoceros.
By the test-results of the foregoing description 8～10 as can be known, when cultured continuously, culture condition (P/N 〉=0.12, Si/N 〉=0.36) is all set up indoor and outdoor.Thereby, compare with existing cultural method, even during cultured continuously, also can high cell concentration (indoorly be about 6.0 * 10 13Cell/m 3, outdoorly be about 5.0 * 10 13Cell/m 3) cultivate Chaetoceros continuously and stably.In addition, indoor or outdoors no matter, the pH of nutrient solution 4 all maintain in the scope more than 6.4 or 6.4, below 8.5 or 8.5.
Need to prove because Chaetoceros is slow to the absorption speed of nitrogen and silicon, therefore in embodiment 8～10, before the initial stage concentration determination of nutrient solution 4,6 is not defined as the Chaetoceros inoculation, for example also can after just carrying out Chaetoceros inoculation back (with reference to Figure 32) or inoculating half, carry out.
In addition, among the embodiment 8～10, to be actually used in the test nutrient solution of cultivating Chaetoceros is object, measure nitrogen consumption, phosphorus consumption, silicon consumption by the real time, but can be object with nutrient solution also to have the consumption mensuration of forming with the nutrient solution identical component with test, the inoculation Chaetoceros is measured nitrogen consumption, phosphorus consumption, silicon consumption in advance.
Its reason is as follows.With regard to the Chaetoceros of per unit cell with regard to the nitrogen amount (NCOM) of time per unit consumption, phosphorus amount (PCOM), the silicon amount (SiCOM), time test and resolve that the value of PCOM/NCOM and SiCO/NCOM often remains necessarily in the actual as can be known culture experiment thus.Thereby measure nitrogen consumption, phosphorus consumption, silicon consumption by measuring in advance with nutrient solution, obtain the P of the diatom of cultivating by consumption ALL/ N ALLAnd Si ALL/ N ALLEven, pipeline time of culture experiment not determination test with nitrogen consumption, phosphorus consumption, the silicon consumption of nutrient solution, as long as supply adjusted topdressing of P/N and Si/N respectively and use nutrient solution, so that itself and the P that obtains in advance ALL/ N ALLAnd Si ALL/ N ALLUnanimity just can be simplified culturing step, forms by the nutrient solution of optimal dose and topdresses.
For example, because the Chaetoceros amount inoculated in nutrient solution equals among the embodiment 10 in the nutrient solution 4 under the situation of inoculum size P ALL/ N ALL, Si ALL/ N ALLValue equal 0.143,0.402 (with reference to Figure 27) respectively, therefore the P/N and the Si/N value of topdressing with nutrient solution equal 0.143,0.402 respectively.
In addition, in embodiment 8～10, the cell concn of Chaetoceros surpasses about 6.0 * 10 13Cell/m 3The time, will topdress and go into nutrient solution 4, but be not limited to this with the nutrient solution supply, can set the cell concn of Chaetoceros arbitrarily.
The advantage of cultural method of the present invention is as described below:
Because therefore the increase of the cultivation amount of per unit volume Chaetoceros is compared with existing cultural method, workload, cultivation interval, keeping interval and transportation cost reduce.For example, for the minimizing of workload, be that the amount that drops into the Chaetoceros nutrient solution in the tank of aquatic products seedling of bait reduces having with the Chaetoceros.
When cultivating beginning,, therefore can predict the required time of cell concn that arrives specified amount owing to, can adjust the cell concn when cultivating beginning by changing the inoculum size of Chaetoceros.
In above-mentioned the present embodiment, the diatom that is seeded in the nutrient solution is a Chaetoceros, but is not limited thereto, and also can be the diatom that is categorized as other genus.
For example, when cultivating the diatom (hereinafter referred to as brown algae) that is categorized as brown algae (Phaeodactylum) genus, prepare to have the nutrient solution 10 that one-tenth shown in Figure 34 is grouped into.The composition that need to prove nutrient solution 10 is identical with nutrient solution 4, P/N=0.12, Si/N=0.36.In addition, the composition of nutrient solution 11 one-tenth that consists of the known nutrient solution that brown algae uses in cultivating is grouped into P/N=1.29, Si/N=0 (promptly not adding silicon).
(embodiment 11)
With nutrient solution 10 (1.5 * 10 -3m 3) put into the laboratory with flat culturing bottle 11 (capacity 1.5 * 10 -3m 3), interpolation sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, the brown algae of inoculation specified amount in nutrient solution 10, the condition roughly the same with embodiment 1 (culture-liquid temp: about 25 ℃～about 35 ℃, photon amount: about 200 μ mol/m 2/ s carries out aeration-agitation by the air that has mixed 3% carbonic acid gas) under, brown algae cultivated.Measure the time variation of brown algae cell concn in the nutrient solution 10 and the time variation of pH respectively.
(comparative example)
With nutrient solution 11 (1.5 * 10 -3m 3) join in the flat culturing bottle 11, interpolation sodium bicarbonate to the mass concentration of sodium bicarbonate is 1.0 * 10 3Ppm.Then, inoculation and the brown algae of embodiment 11 in nutrient solution 11 with amount.Next, the condition roughly the same with embodiment 1 (culture-liquid temp: about 25 ℃～about 35 ℃, photon amount: about 200 μ mol/m 2/ s carries out aeration-agitation by the air that has mixed 3% carbonic acid gas) under, brown algae cultivated.Measure the time variation of brown algae cell concn in the nutrient solution 11 and the time variation of pH respectively.
Figure 35 illustrates the graphic representation that the brown algae cell concn time changes in embodiment 11 and the comparative example.M1 shown in Figure 35, M2 represent the brown algae cell concn of cultivation in the nutrient solution 10,11 respectively.
Figure 36 illustrates the graphic representation that the pH time changes in embodiment 11 and the comparative example.N1 shown in Figure 36, N2 represent the pH in the nutrient solution 10,11 respectively.
Test-results is as can be known thus:
The cell concn of brown algae surpasses about 1.2 * 10 in the nutrient solution 10 14Cell/m 3The cell concn of brown algae does not reach about 4.0 * 10 in the comparative example 13Cell/m 3
PH in the nutrient solution 10 maintains the scope more than 6.4 or 6.4, below 8.5 or 8.5, and the pH in the comparative example maintains in the scope more than about 6.4 or 6.4, below about 8.5 or 8.5.
By the test-results of the foregoing description 11 as can be known, cultivate brown algaes, compare with existing nutrient solution 11 by using nutrient solution 10, can high cell concentration and stably cultivate brown algae.
Thereby, identical with the situation of Chaetoceros, compare with existing cultural method, can reduce workload, culture space, keeping space and transportation cost.
Need to prove in embodiment 1～11,, add sodium bicarbonate as the pH regulator agent of nutrient solution.But, except sodium bicarbonate, also can add the supercarbonate that contains sodium 1 valency basic metal [Li (lithium), K (potassium), Rb (rubidium), Cs (caesium) etc.] in addition, so that contain supercarbonate in the nutrient solution.In addition,, also can add divalent basic metal, so that contain alkali-metal supercarbonate in the nutrient solution as the pH regulator agent.
In addition, in embodiment 1～11,, use the air that has mixed 3% carbonic acid gas as the gas of aeration-agitation nutrient solution, but, also can be the gas of compositions such as the oxygen that added specified amount, carbonic acid gas so long as life is kept necessary composition in the nutrient solution.
And, in the present embodiment, modulate P/N and Si/N in the nutrient solution of cultivating diatom respectively, but the present invention is not limited thereto formation, as long as it is just passable to regulate Si/N at least.
The present invention is not limited to above-mentioned embodiment and embodiment, in the scope that does not break away from purport of the present invention, can suitably revise the back and implement.

Claims (41)

1, a kind of liquid that contains diatom is the liquid that is used to cultivate diatom, it is characterized in that, described liquid contains diatom, silicon and nitrogen, and silicon/nitrogen (mass concentration ratio of described silicon and described nitrogen) is set at the value greater than 0.18.
2, the liquid that contains diatom as claimed in claim 1 is characterized in that, the pH of described liquid is more than 6.4 or 6.4.
3, the liquid that contains diatom as claimed in claim 1 is characterized in that, described liquid contains supercarbonate.
4, the liquid that contains diatom as claimed in claim 1 is characterized in that, is to be roughly consistent with the silicon consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the silicon in the solution before the described diatom of inoculation/nitrogen decision.
5, the liquid that contains diatom as claimed in claim 1 is characterized in that, the frustule concentration of being cultivated is 4.0 * 10 13Cell/m 3Or 4.0 * 10 13Cell/m 3More than.
6, the liquid that contains diatom as claimed in claim 1 is characterized in that, described diatom is categorized as Chaetoceros and belongs to.
7, the liquid that contains diatom as claimed in claim 1 is characterized in that, described diatom is categorized as brown algae (Phaeodactylum) and belongs to.
8, a kind of liquid that contains diatom, be the liquid that is used to cultivate diatom, it is characterized in that described liquid contains diatom, phosphorus, silicon and nitrogen, phosphorus/nitrogen (mass concentration ratio of described phosphorus and described nitrogen), and silicon/nitrogen (mass concentration ratio of described silicon and described nitrogen) be set at value more than 0.1 or 0.1.
9, the liquid that contains diatom as claimed in claim 8 is characterized in that, the pH of described liquid is more than 6.4 or 6.4.
10, the liquid that contains diatom as claimed in claim 8 is characterized in that, described liquid contains supercarbonate.
11, the liquid that contains diatom as claimed in claim 8 is characterized in that, is to be roughly consistent with the silicon consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the silicon in the solution before the described diatom of inoculation/nitrogen decision.
12, the liquid that contains diatom as claimed in claim 8 is characterized in that, is to be roughly consistent with the phosphorus consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the phosphorus in the solution before the described diatom of inoculation/nitrogen decision.
13, the liquid that contains diatom as claimed in claim 8 is characterized in that, the frustule concentration of being cultivated is 4.0 * 10 13Cell/m 3Or 4.0 * 10 13Cell/m 3More than.
14, the liquid that contains diatom as claimed in claim 8 is characterized in that, described diatom is categorized as Chaetoceros and belongs to.
15, the liquid that contains diatom as claimed in claim 8 is characterized in that, described diatom is categorized as brown algae (Phaeodactylum) and belongs to.
16, a kind of liquid that contains diatom is the liquid that is used to cultivate diatom, it is characterized in that, described liquid contains diatom, phosphorus, silicon and nitrogen, and pH is more than 6.4 or 6.4, below 8.4 or 8.4.
17, the liquid that contains diatom as claimed in claim 16 is characterized in that, phosphorus/nitrogen (mass concentration ratio of described phosphorus and described nitrogen) is set at more than 0.06 or 0.06, and silicon/nitrogen (mass concentration ratio of described silicon and described nitrogen) is set at more than 0.18 or 0.18.
18, the liquid that contains diatom as claimed in claim 16 is characterized in that, described liquid contains supercarbonate.
19, the liquid that contains diatom as claimed in claim 16 is characterized in that, is to be roughly consistent with the silicon consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the silicon in the solution before the described diatom of inoculation/nitrogen decision.
20, the liquid that contains diatom as claimed in claim 16 is characterized in that, is to be roughly consistent with the phosphorus consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the phosphorus in the solution before the described diatom of inoculation/nitrogen decision.
21, the liquid that contains diatom as claimed in claim 16 is characterized in that, described diatom is categorized as Chaetoceros and belongs to.
22, the liquid that contains diatom as claimed in claim 16 is characterized in that, described diatom is categorized as brown algae (Phaeodactylum) and belongs to.
23, a kind of diatom is characterized in that, described diatom is cultivated in following liquid and forms, and described liquid contains silicon and nitrogen, and silicon/nitrogen (mass concentration ratio of described silicon and described nitrogen) is set at the value above 0.18.
24, a kind of diatom, it is characterized in that, described diatom is cultivated in following liquid and forms, and described liquid contains phosphorus, nitrogen and silicon, and phosphorus/nitrogen (mass concentration ratio of described phosphorus and described nitrogen) and silicon/nitrogen (mass concentration ratio of described silicon and described nitrogen) are set at the value more than 0.1 or 0.1 respectively.
25, a kind of diatom is characterized in that, described diatom is cultivated in following liquid and forms, and described liquid contains phosphorus, nitrogen and silicon, and pH is set in the scope more than 6.4 or 6.4, below 8.4 or 8.4.
26, the cultural method of a kind of diatom is to use the diatom nutrient solution that contains nitrogen and silicon to cultivate the diatom cultural method of diatom, it is characterized in that described method comprises the steps:
Measure respectively in the described liquid the determination step in advance of initial stage concentration of nitrogenous initial stage concentration and silicon;
The inoculation step of the described diatom of inoculation in described liquid;
Measure determination step before the supply of concentration before the supply of concentration and described silicon before the supply of described nitrogen respectively;
Obtain the concentration difference determination step of nitrogen concentration poor (concentration is poor before the initial stage concentration of described nitrogen and the supply) and silicon concentration poor (concentration is poor before the initial stage concentration of described silicon and the supply) respectively;
Poor based on described nitrogen concentration difference and described silicon concentration, determine the nitrogen amount and the silicon amount of supply in described liquid respectively, and its supply is gone in the described liquid, so that itself and the consistent supply step of ratio of the initial stage concentration of the initial stage concentration of described nitrogen and described silicon.
27, the cultural method of a kind of diatom is to use the diatom nutrient solution that contains nitrogen and silicon to cultivate the diatom cultural method of diatom, it is characterized in that described method comprises the steps:
Measure determination step before the supply of concentration before the supply of concentration and described silicon before the supply of described nitrogen respectively;
Obtain the concentration difference determination step of nitrogen concentration poor (concentration is poor before the normality of described nitrogen and the supply) and silicon concentration poor (concentration is poor before the normality of described silicon and the supply) respectively;
Poor based on described nitrogen concentration difference and described silicon concentration, determine the nitrogen amount and the silicon amount of supply in described liquid respectively, and its supply is gone in the described liquid, so that itself and the consistent supply step of ratio of the normality of the normality of described nitrogen and described silicon.
28, diatom cultural method as claimed in claim 27 is characterized in that, described liquid also contains phosphorus; Before supply, in the determination step, measure the preceding concentration of supply of described phosphorus; In the concentration difference determination step, obtain phosphorus concentration poor (concentration is poor before the normality of described phosphorus and the supply); In the supply step, poor based on described phosphorus concentration, the phosphorus amount of decision supply in described liquid, and its supply gone in the described liquid, so that its ratio with the normality of the normality of described nitrogen and described phosphorus is consistent.
29, diatom cultural method as claimed in claim 27 is characterized in that, the cultivation limit of described diatom feeds the gas limit and carries out in described liquid.
30, diatom cultural method as claimed in claim 27 is characterized in that, is to be roughly consistent with the silicon consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the silicon in the solution before the described diatom of inoculation/nitrogen decision.
31, diatom cultural method as claimed in claim 28 is characterized in that, is to be roughly consistent with the phosphorus consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the phosphorus in the solution before the described diatom of inoculation/nitrogen decision.
32, the cultural method of a kind of diatom is to use the diatom nutrient solution that contains nitrogen and silicon to cultivate the diatom cultural method of diatom, it is characterized in that described method comprises the steps:
Be determined in the liquid of having inoculated described diatom the determination step in advance of initial stage concentration of nitrogenous initial stage concentration and silicon;
Measure determination step before the supply of concentration before the supply of concentration and described silicon before the supply of described nitrogen respectively;
Obtain the concentration difference determination step of nitrogen concentration poor (concentration is poor before the initial stage concentration of described nitrogen and the supply) and silicon concentration poor (concentration is poor before the initial stage concentration of described silicon and the supply) respectively;
Poor based on described nitrogen concentration difference and described silicon concentration, determine the nitrogen amount and the silicon amount of supply in described liquid respectively, and its supply is gone in the described liquid, so that itself and the consistent supply step of ratio of the initial stage concentration of the initial stage concentration of described nitrogen and described silicon.
33, diatom cultural method as claimed in claim 32 is characterized in that, described liquid also contains phosphorus; In determination step in advance, measure the initial stage concentration of described phosphorus; Before supply, in the determination step, measure the preceding concentration of supply of described phosphorus; In the concentration difference determination step, obtain phosphorus concentration poor (concentration is poor before the initial stage concentration of described phosphorus and the supply); In the supply step, poor based on described phosphorus concentration, the phosphorus amount of decision supply in described liquid, and its supply gone in the described liquid is so that its ratio with the normality of the normality of described nitrogen and described phosphorus is consistent.
34, diatom cultural method as claimed in claim 32 is characterized in that, the cultivation limit of described diatom feeds the gas limit and carries out in described liquid.
35, diatom cultural method as claimed in claim 32 is characterized in that, is to be roughly consistent with the silicon consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the silicon in the solution before the described diatom of inoculation/nitrogen decision.
36, diatom cultural method as claimed in claim 33 is characterized in that, is to be roughly consistent with the phosphorus consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the phosphorus in the solution before the described diatom of inoculation/nitrogen decision.
37, the cultural method of a kind of diatom is to use the diatom nutrient solution that contains nitrogen and silicon to cultivate the diatom cultural method of diatom, it is characterized in that described method comprises the steps:
Measure with in the liquid at the consumption of having inoculated described diatom, measure the nitrogen consumption of described diatom consumption and the determination step in advance of silicon consumption respectively;
The inoculation step of the described diatom of inoculation in described liquid;
Measure the determination step of the cell concn of described diatom;
Described cell concn surpasses under the situation of specified amount, and modulation is topdressed with liquid so that the consistent modulation step of ratio of the consumption of the consumption of the ratio of nitrogen and silicon and described nitrogen and described silicon;
Synthetic described topdressing gone into supply step in the described nutrient solution with the liquid supply.
38, diatom cultural method as claimed in claim 37 is characterized in that, described nutrient solution also contains phosphorus; In determination step in advance, measure described consumption measure with in the liquid by the phosphorus consumption of described diatom consumption; In modulation step, modulation is topdressed and is used liquid, so that the ratio of the consumption of the consumption of the ratio of nitrogen and silicon and described nitrogen and described phosphorus is consistent.
39, diatom cultural method as claimed in claim 37 is characterized in that, the cultivation limit of described diatom feeds the gas limit and carries out in described liquid.
40, diatom cultural method as claimed in claim 37 is characterized in that, is to be roughly consistent with the silicon consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the silicon in the solution before the described diatom of inoculation/nitrogen decision.
41, diatom cultural method as claimed in claim 38 is characterized in that, is to be roughly consistent with the phosphorus consumption of the described diatom of measuring in advance and the ratio of nitrogen consumption with the phosphorus in the solution before the described diatom of inoculation/nitrogen decision.
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CN109152341A (en) * 2016-02-24 2019-01-04 南特大学 By means of CO2Source is come the method for cultivating photosynthetic organism
CN107673874A (en) * 2017-11-10 2018-02-09 林东亨 Diatomeae liquid compound fertilizer and preparation method thereof
CN109511580A (en) * 2018-10-30 2019-03-26 南宁学院 A kind of ecological efficient cultural method of mudskipper

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