CN112574889A - Method for rapidly expanding and propagating bait microalgae biomass by using polyculture culture - Google Patents
Method for rapidly expanding and propagating bait microalgae biomass by using polyculture culture Download PDFInfo
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Abstract
The invention provides a culture method for rapid propagation of bait microalgae, which is characterized in that an organic carbon source is added into an f/2 culture medium to culture the bait microalgae; the bait microalgae are respectively pseudominitype Alternaria hainanensis, chlorella or Chaetoceros sp; preferably, the organic carbon source added to the f/2 culture medium is glycerol, acetic acid or glucose; wherein the concentration of the organic carbon source is 0.5-5.0g.L‑1. The method can effectively improve the nutritive value of the bait microalgae, can improve the propagation speed of the bait microalgae, reduce the production cost of the bait microalgae, improve the economy of shellfish culture, and provide technical support for wide low-cost application of the microalgae shellfish bait.
Description
Technical Field
The invention belongs to the technical field of bait microalgae propagation culture and production of active products thereof, and particularly relates to a method for rapidly propagating bait microalgae biomass by using polyculture culture.
Background
Bivalves are an important economic support for the marine organism industry and have an important share in the world fishery production. Living body microalgae have been used in bivalve shellfish farming processes and are considered to be the most suitable feed for growth of shellfish larvae and juvenile mollusks. Microalgae is a bottom organism of a food chain of a water ecological system, and part of algae cells are rich in oil, polysaccharide, polyunsaturated fatty acid and the like, have high nutritive value and are the key for improving the survival rate of the bivalve shellfish larvae.
At present, some bait microalgae, such as pseudonandina hainanensis, chlorella, chaetoceros and the like, become one of the most popular microalgae in the process of culturing bivalve shellfish due to the characteristics of proper size, rich nutrition and the like. However, the slow cell growth and long generation time of the bait algae in the actual culture process are not enough to meet the requirement of rapid growth of the shellfish. However, the conventional methods for improving the biological yield of microalgae, such as optimization of culture conditions and improvement of culture devices, cannot effectively improve the growth of the conventional bait microalgae. How to rapidly and efficiently propagate bait microalgae is the key for promoting the growth of shellfish and reducing the shellfish cultivation cost.
Generally, microalgae utilize sunlight (natural light) and air CO in the actual aquaculture water2From autotrophic growth. The microalgae is illuminated and CO is generated in the autotrophic growth process2The factors have great influence, which is the main reason that the bait microalgae in the naturally cultivated water area grow slowly. Therefore, changing the nutritional mode of the bait microalgae is a possible way to improve the growth and nutritional value of the bait microalgae.
Disclosure of Invention
The invention aims to provide a method for rapidly expanding and propagating bait microalgae biomass by using mixed culture, namely a mixed nutrition mode of adding an organic carbon source into a culture medium, so that the expanding and propagating speed of the bait microalgae is improved, the culture cost of the bait microalgae is further reduced, the economy of shellfish culture is improved, and technical support is provided for the wide application of bait microalgae shellfish seedling culture.
The invention firstly provides a method for culturing bait microalgae, which is characterized in that an organic carbon source is added into an f/2 culture medium to culture the bait microalgae;
the bait microalgae are respectively pseudominitype Alternaria hainanensis, chlorella or Chaetoceros sp;
preferably, the organic carbon source added to the f/2 medium is glucose, acetic acid or glycerol;
wherein the concentration of the organic carbon source is 0.5-5.0g.L-1；
As a specific description of the examples, when chlorella is cultured, 5.0g.L of chlorella is added to f/2 medium-1Acetic acid of (a);
when culturing the pseudo-micro-Alhainanensis or Chaetoceros, 5.0g.L of the culture medium is added into f/2-1As an organic carbon source;
as a specific description of the embodiment, the bait microalgae is cultured in mixed nutrition at 25 ℃ and the illumination intensity of 80 mu mol.m-2.s-1Photoperiod 12 h: in 12h of liquid culture.
The method can effectively improve the nutritive value of the bait microalgae, can improve the propagation speed of the bait microalgae, reduce the production cost of the bait microalgae, improve the economy of shellfish culture, and provide technical support for wide low-cost application of the microalgae shellfish bait.
Drawings
FIG. 1: an experimental technical route chart for bait microalgae mixed nutrition culture screening;
FIG. 2: 3, a mixed culture diagram of different organic carbon sources of the bait microalgae;
FIG. 3: 3, a mixed culture diagram of different organic carbon source concentrations of bait algae;
FIG. 4: 3 bait microalgae optimal organic carbon source and biochemical component change diagram under concentration.
Detailed Description
The present invention will be described in detail below with reference to examples and the accompanying drawings.
Example 1: comparison of cultivation effects of different organic carbon sources of bait microalgae
Respectively culturing 3 common bait microalgae Chlorella, Thalassiosira pseudonana, and Chaetoceros in f/2 culture medium with illumination intensity of 80 μmol.m-2.s-1Strip at 25 DEG CThe culture was carried out in a flask with a mixed nutrient solution (FIG. 1) and the concentration of the solution was 2.0g.L in the f/2 medium-1The organic carbon sources of (a) glucose, acetic acid and glycerol. The composition ratio of f/2 medium was as follows (mg.L)-1): 75 parts of sodium nitrate, 5 parts of monobasic sodium phosphate monohydrate, 30 parts of sodium silicate nonahydrate, 3.15 parts of ferric chloride hexahydrate, 4.36 parts of disodium ethylenediamine tetraacetic acid dihydrate, 0.0098 part of copper sulfate pentahydrate, 0.0063 part of sodium molybdate dihydrate, 0.022 part of zinc sulfate heptahydrate, 0.01 part of cobalt chloride hexahydrate, 0.18 part of manganese chloride tetrahydrate, and vitamin B10.1, 0.0005 of vitamin H, and vitamin B12 0.0005。
The volume of the triangular flask used for the bait microalgae liquid suspension culture experiment is 250mL, and the result is shown in FIG. 2 after 11 days of culture: for chlorella, it grows well under organic carbon source acetic acid with biomass of 135.7 × 105cells/ml, better than the f/2 normal medium value of 80X 105cells/ml. For the Thalassiosira pseudonana, it grows well under organic carbon source glycerol with biomass of 25.8X 105cells/ml, better than f/2 normal medium value of 11.5X 105cells/ml. For Chaetoceros, algal cells also grow well under organic carbon source glycerol with biomass of 53.8X 105cells/ml, 39.6X 10 better than f/2 normal medium5cells/ml。
Example 2: optimized screening of different carbon source concentrations under mixed nutrient condition
According to the experimental scheme of figure 1, 3 bait microalgae Chlorella, Thalassiosira pseudonana, Chaetoceros are irradiated under the same illumination intensity (80 μmol. m)-2.s-1) Culturing under the temperature condition (25 ℃); organic carbon sources of acetic acid, glycerol and glycerol were added to the f/2 normal medium, respectively. The concentrations of the three organic carbon sources are respectively set to be 0.5, 1.0, 2.0 and 5.0g.L-1The results of 11 days of culture under the same conditions as in example 1 are shown in FIG. 3. The result showed that the concentration of chlorella in acetic acid was 5.0g.L-1Has good growth effect and biomass of 141.7 multiplied by 105cells/ml, 84X 10 better than f/2 normal medium5cells/ml; the concentration of the Thalassiosira pseudonana in the organic carbon source glycerol is 5.0g.L-1The growth is good, the biomass is 70.5 multiplied by 105cells/mlFar superior to the value of 17.2X 10 of f/2 normal medium5cells/ml; chaetoceros has a concentration of 5.0g.L in organic carbon source glycerol-1The growth is good, the biomass is 55.2 multiplied by 105cells/ml, far superior to the f/2 normal medium value of 44.3X 105cells/ml. The 3 bait algae strains are under high-concentration organic carbon source (20 g.L)-1) The growth of algal cells is not correspondingly increased and is not described herein.
Example 3: optimum organic carbon source and biochemical components of algal cells and main fatty acid change under concentration of bait microalgae
3 bait microalgae Chlorella, Thalassiosira pseudonana and Chaetoceros were added to 5.0g.L of the optimum organic carbon source under the conditions described in examples 1 and 2-1Acetic acid, 5.0g.L-1Glycerol and 5.0g.L-1Culturing in f/2 normal culture medium of glycerol for 11 days, and measuring contents of oil, saccharide, protein and main fatty acid of algae cell. The results are shown in fig. 4, and the polysaccharide and the oil and fat components of the 3 bait algae strains are superior to the values under the corresponding normal culture medium conditions under the organic carbon source condition.
More importantly, under the condition of the dominant organic carbon source and concentration, the content of the components of polyunsaturated fatty acids C20:5 and C22:6 in the pseudo-micro-sea chain algae is respectively increased by 22 percent and 31 percent compared with the corresponding value in a normal f/2 culture medium; for Chaetoceros, the content of the polyunsaturated fatty acid C18:2 component was increased by about 17% over the corresponding value in the normal f/2 medium.
The results of example 1, example 2 and example 3 show that the method of the invention can not only promote the growth of bait algae, but also improve the contents of high-value products such as algal cell polysaccharides, grease, partial polyunsaturated fatty acids and the like, and improve the nutritional value of the bait microalgae.
Claims (7)
1. A method for culturing bait microalgae is characterized in that organic carbon sources are added into an f/2 culture medium to culture the bait microalgae; the bait microalgae is pseudonandina hainanensis, chlorella or chaetoceros.
2. The method of claim 1, wherein the organic carbon source is added at a concentration of 0.5 to 5.0g.L-1。
3. The method of claim 1, wherein the organic carbon source is added at a concentration of 5.0g.L-1。
4. The method of any one of claims 1 to 3, wherein the organic carbon source is glucose, acetic acid or glycerol.
5. The method according to claim 1, wherein 5.0g.L of chlorella is added to f/2 medium during the culture of chlorella-1Acetic acid (2).
6. The method according to claim 1, wherein 5.0g.L of the culture medium f/2 is added to the culture medium during the cultivation of the Thalassiosira pseudonana or Chaetoceros sp-1As an organic carbon source.
7. The method according to any one of claims 1 to 6, wherein the method is carried out at a temperature of 25 ℃ and an illumination intensity of 80 μmol-2.s-1Photoperiod 12 h: culturing is carried out under the condition of 12 h.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106591131A (en) * | 2015-10-20 | 2017-04-26 | 青岛力天宏泰新能源科技有限公司 | Heterotrophic culture medium used for large-scale cultivation of marine microalgae |
| CN109825438A (en) * | 2019-03-01 | 2019-05-31 | 华东理工大学 | The method for cultivating Rhodophyta single-cell sea microalgae production bioactive ingredients |
| CN110923185A (en) * | 2018-09-19 | 2020-03-27 | 中国科学院青岛生物能源与过程研究所 | Culture method for improving oil content of microalgae cells |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106591131A (en) * | 2015-10-20 | 2017-04-26 | 青岛力天宏泰新能源科技有限公司 | Heterotrophic culture medium used for large-scale cultivation of marine microalgae |
| CN110923185A (en) * | 2018-09-19 | 2020-03-27 | 中国科学院青岛生物能源与过程研究所 | Culture method for improving oil content of microalgae cells |
| CN109825438A (en) * | 2019-03-01 | 2019-05-31 | 华东理工大学 | The method for cultivating Rhodophyta single-cell sea microalgae production bioactive ingredients |
Non-Patent Citations (1)
| Title |
|---|
| 农业部工人技术培训教材编审委员会编: "《海洋生物饵料培养》", 30 May 1995, 中国农业出版社 * |
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