CN111172096B - Production process for high-density culture of heteroglena - Google Patents
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Abstract
The invention belongs to the technical field of algae microorganism culture, and discloses a production process for high-density culture of heteroglena, which comprises the following steps: selecting vigorous and pollution-free Isoglena, inoculating into culture pond containing Isoglena culture solution, controlling temperature at 23-26 deg.C, irradiating with red light at light-dark time ratio of 10-14:14-10, introducing air flow of 0.4-0.6vvm, and culturing for 7-9 d. The production process can improve the growth amount of the heteroglena.
Description
Technical Field
The invention belongs to the technical field of algae microorganism culture, and particularly relates to a production process for high-density culture of heteroglena.
Background
Plankton plays an important role in aquaculture, and is indispensable for maintaining the normal functions of water ecosystem and stabilizing pond water environment. In the continuous development process of the species group, on one hand, the microalgae provide sufficient dissolved oxygen for the water body through photosynthesis, on the other hand, organic pollution and other harmful substances in the aquaculture water body are reduced and eliminated, the virtuous cycle of the aquaculture ecological system is kept, and the purpose of improving water quality is achieved. Therefore, the algae can be reasonably utilized in the fry cultivation and water quality management of aquaculture, and the effect of achieving twice the result with half the effort can be achieved. The microalgae is rich in nutrient substances, and many algae contain high protein, and also contain vitamins and trace elements which are necessary for maintaining the growth of aquatic product seedlings, so that the microalgae has an obvious promotion effect on various physiological functions of the seedlings. For example, the vitamin E has the effects of promoting phagocytosis of macrophages of aquatic animals, improving complement activity and enhancing the immunity of larvae of the aquatic animals; the addition of anabaena powder in the compound feed for prawns can obviously improve the growth speed and disease resistance of prawns. Therefore, the excellent algae species such as anabaena, chlorella and the like have good application prospect in the cultivation of fish and shrimp fries.
The unicellular algae in a certain proportion is added into the compound feed to serve as bait in the early stage of fish and shrimp seedling raising, the survival rate of seedlings is higher, and the seedlings can be raised with high and stable yield. Therefore, in many areas, the larvae are cultivated by adding non-biological baits to unicellular algae. Different algae species have certain differences in the content of nutritional indexes. The mixed regulation and control mode of various algae is beneficial to the balanced supply of various nutrient elements, thereby ensuring the growth, development and smooth metamorphosis of the larva. The algae species are complicated, the quantity is large, and the culture methods of different algae are different and cannot be used indiscriminately. At present, only a few kinds of bait algae which are widely applied in aquaculture are provided, wherein chlorella (the first artificially cultured microalgae in China), dunaliella salina, chaetoceros sp, phaeodactylum tricornutum, equilateral chrysophyceae and the like exist, and in addition, skeletonema costatum, isoproteinella, Platymonas fasciata and Nitzschia closterium also gradually start to be cultured.
Besides being used as direct bait, the microalgae has another important function of being used for feeding secondary baits such as rotifers, artemia, copepods, cladocerans and the like, and can obviously enhance the content of polyunsaturated fatty acids and various vitamins contained in organisms of the secondary baits so as to meet the requirement of aquatic animal larvae on high-quality secondary baits. The wide application of the excellent algae species powerfully promotes the development of the seedling industry and also promotes the progress of the seedling raising technology. With the increasing attention paid to the concept of healthy cultivation, it is expected that the algae will certainly play more and more important roles in the offspring seed cultivation of aquatic economic animals in the future.
The heteroglea is good bait algae, but the research on artificial high-density culture is less, the difference between the physical and chemical properties of different algae and the culture conditions is large, and the culture conditions of other algae cannot be applied. Only a few reports exist at present, and CN 106591137A discloses an isoglena culture medium composition, which comprises the following components: KNO 3: 300400 mg/L; KH2PO 4:10 mg/L; feso 4.7h2o: 2.5 mg/L; na2 EDTA: 10 mg/L; MnSO 4: 0.25 g/L: VB 1: 6 mg/L; VB 12: 50 ug/L; additive: 0.53 g/L; the plant growth regulator prepared from 6-BA and naphthylacetic acid has the advantages of high growth rate and high cell density when culturing the heteroglea. CN 104130945A utilizes leather factory waste water to cultivate heteroglena, can realize the reuse of useful substances in the leather factory waste water and can improve the growth speed and the nutrient content of the heteroglena, thereby achieving the double effects of protecting the environment and producing biological resources, having simple, convenient and quick operation, low cost, no pollution, high social, economic and ecological benefits, solving the problem of waste water pollution of the leather factory, solving the defects of slow growth, low yield, poor benefits, easy pollution and low nutrient content of the heteroglena, and realizing the production culture of the heteroglena. However, the method has the defects of high cost, unclean water body, easy carrying of pollutants, low density, poor environmental friendliness and the like.
Disclosure of Invention
In order to solve the technical defects in the prior art, the invention provides a production process for culturing heteroglena at high density.
The invention is realized by the following technical scheme:
a production process for culturing alloglena at high density comprises the following steps:
selecting vigorous and pollution-free Isoglena, inoculating into culture pond containing Isoglena culture solution, controlling temperature at 23-26 deg.C, irradiating with red light at light-dark time ratio of 10-14:14-10, introducing air flow of 0.4-0.6vvm, and culturing for 7-9 d.
Further, the isopulegolic algae culture solution is prepared according to the following steps: ammonium bicarbonate and brassinolide were added to the f/2 medium.
Preferably, the concentration of the ammonium bicarbonate is 30-50 mg/L.
Preferably, the concentration of the brassinolide is 0.02-0.03 mg/L.
Furthermore, the salinity is controlled to be 22-26 by feeding sodium chloride aqueous solution with the concentration of 2-3mol/L in the whole culture process.
Further, the inoculation density was 500 ten thousand per ml.
Further, the wavelength of the red light is between 720 and 740 nm.
Preferably, the light intensity of the red light is 45-55 [ mu ] mol/(m)2.S）。
The beneficial effects of the invention mainly comprise the following aspects:
compared with the conventional cold light white, the infrared light with a certain wavelength can more effectively stimulate the photoreaction system of the heteroglea and stimulate the heteroglea to divide and proliferate more quickly, thereby realizing high-density culture in a short time.
After inoculation, a short delay period appears, the culture reaches a higher increase for 3d until 7-9d, then a stationary period is entered, and the increase rate is obviously reduced, so that the control of the culture period to be 7-9 days is more suitable.
The f/2 culture medium is suitable for most algae to grow, but is not the optimal culture medium for most algae, and on the basis of the f/2 culture medium, the ammonium bicarbonate is added, so that the absorption of nitrogen by the alloglena can be promoted, carbon dioxide can be provided for the alloglena to utilize, the growth of algae cells is stimulated, the biomass is improved, and no environmental pollution substances are generated; the brassinolide can obviously promote the cell division of the heteroglea under the red light condition, maintain the rapid proliferation of the heteroglea and improve the growth amount of the algae cells.
Drawings
FIG. 1: the effect of ammonium bicarbonate on the biomass of heteroglena;
FIG. 2: the effect of brassinolide on the biomass of heteroglena;
FIG. 3: influence of different wavelengths of light on biomass of the heteroglena.
Detailed Description
Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the products and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations and modifications, or appropriate alterations and combinations, of the products and methods described herein may be made and utilized without departing from the spirit, scope, and spirit of the invention. For a further understanding of the present invention, reference will now be made in detail to the following examples.
Example 1
A production process for culturing alloglena at high density comprises the following steps:
preparing an isopulegolic algae culture solution: the f/2 culture medium is improved, and the specific steps are as follows: adding ammonium bicarbonate and brassinolide into the f/2 culture medium, and controlling the concentrations of the ammonium bicarbonate and the brassinolide to be 50mg/L and 0.03mg/L respectively;
high-density culture: selecting vigorous and pollution-free Isoglena, inoculating into culture pond containing Isoglena culture solution, inoculating at density of 500 ten thousand per ml, controlling temperature at 25 deg.C, irradiating with red light with electromagnetic radiation of 720nm wavelength, and light intensity of 45 μmol/(m)2S), the light-dark time ratio is 12:12, the air flow is introduced to be 0.5vvm, and the culture is carried out for 9 d; the salinity is controlled to be 23 by feeding sodium chloride aqueous solution with the concentration of 2mol/L in the whole culture process.
Example 2
A production process for culturing alloglena at high density comprises the following steps:
preparing an isopulegolic algae culture solution: the f/2 culture medium is improved, and the specific steps are as follows: adding ammonium bicarbonate and brassinolide into the f/2 culture medium, and controlling the concentrations of the ammonium bicarbonate and the brassinolide to be 30mg/L and 0.02mg/L respectively;
high-density culture: selecting vigorous and pollution-free Isoglena, inoculating into culture pond containing Isoglena culture solution at an inoculation density of 500 ten thousand per ml, controlling temperature at 25 deg.C, irradiating with red light having electromagnetic radiation of 740nm wavelength, and light intensity of 55 μmol/(m)2S), the light-dark time ratio is 14:10, the air flow is introduced to be 0.6vvm, and the culture is carried out for 8 d; during the whole culture process, the salinity is controlled to be 25 by feeding sodium chloride aqueous solution with the concentration of 3 mol/L.
Example 3
A production process for culturing alloglena at high density comprises the following steps:
preparing an isopulegolic algae culture solution: the f/2 culture medium is improved, and the specific steps are as follows: adding ammonium bicarbonate and brassinolide into the f/2 culture medium, wherein the concentrations of the ammonium bicarbonate and the brassinolide are controlled to be 40mg/L and 0.025mg/L respectively;
high-density culture: selecting vigorous and pollution-free Isoglena, inoculating into annular raceway pond containing Isoglena culture solution at an inoculation density of 500 ten thousand per ml, controlling temperature at 25 deg.C, irradiating with red light having electromagnetic radiation with wavelength of 730nm, and light intensity of 50 μmol/(m) under irradiation of red light2S), light-to-dark time ratio of 10:14, onThe air inlet flow is 0.5vvm, and the culture is carried out for 7 d; the salinity is controlled to be 23 by feeding sodium chloride aqueous solution with the concentration of 2mol/L in the whole culture process.
Comparative example 1
A production process for culturing alloglena comprises the following steps:
high-density culture: selecting vigorous and pollution-free heteroglena, inoculating to a culture pond containing f/2 culture medium, with an inoculation density of 500 ten thousand per ml, a temperature of 25 ℃, irradiation of white light LED (wavelength of 450-2S), the light-dark time ratio is 12:12, the air flow is introduced to be 0.5vvm, and the culture is carried out for 9 d; the salinity is controlled to be 23 by feeding sodium chloride aqueous solution with the concentration of 2mol/L in the whole culture process.
Example 4
1. The f/2 culture medium is an algae conventional culture medium, the invention is improved on the basis of the conventional culture mode of the comparative example 1, and the influence of ammonium bicarbonate and brassinolide on the biomass of the isocollagen is verified. Firstly, adding ammonium bicarbonate on the basis of f/2 culture medium, wherein the addition amount is 0,10,20,30,40,50,60 and 70, and the unit of mg/L is shown in figure 1, when the culture time is increased, the biomass of the heteroglena is rapidly increased, about 9d is close to the maximum value, the culture time is continuously increased, the biomass is slightly increased, but the 9d is selected as the best considering the production period; longitudinally observing that the density of the algae is increased due to the increase of the ammonium bicarbonate concentration, taking the culture time of 9d as an example, the addition amount of 50mg/L of the ammonium bicarbonate concentration enables the biomass of the algae to reach the maximum value, and the ammonium bicarbonate concentration is continuously increased without obvious influence on the biomass of the algae, so the culture time of 9d is selected, and the ammonium bicarbonate concentration is optimally 30-50mg/L, particularly 50 mg/L.
2. Verifying the influence of brassinolide, sodium naphthaleneacetate and indolebutyric acid on the biomass of the isocollagen under the test condition that the ammonium bicarbonate addition concentration is 50mg/L after the culture is carried out for 9 days; the concentration is respectively set to be 0.01,0.02,0.03,0.04,0.05 and 0.06, the unit is mg/L, as shown in figure 2, indolebutyric acid with different concentrations hardly influences the biomass of the heteroglena, and after the sodium naphthaleneacetate reaches 0.05mg/L, the biomass is improved to a small extent, and compared with a group without the indolebutyric acid, the biomass can be improved by only within 5 percent, and the use value is low; the brassinolide with low concentration has obvious promotion effect on the biomass of the heteroglena, the best addition amount of the brassinolide is reached when the addition amount is 0.03mg/L, the concentration is continuously increased, and the biomass of the heteroglena is not obviously improved.
3. The effect of different light sources on the biomass of alloglena.
Selecting light with different wavelengths for testing under the test conditions that the ammonium bicarbonate addition concentration is 50mg/L and the brassinolide addition concentration is 0.03mg/L after culturing for 9d, wherein the wavelength of the white light LED is 460 nm; red light 640nm, 660nm, 680nm, 700nm, 720nm, 740nm and 760 nm; as shown in FIG. 3, compared with the white light, the red light can increase the density of the different kinds of algae, and the influence difference of the red light with different wavelengths on the different kinds of algae is larger, so that the red light with the wavelength of 720-740nm can greatly increase the density of the different kinds of algae, and can increase the density by about 40% compared with the white light.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (2)
1. A production process for culturing alloglena at high density is characterized by comprising the following steps:
selecting vigorous and pollution-free heteroglena, inoculating the heteroglena into a culture pond containing a heteroglena culture solution, controlling the temperature to be 23-26 ℃, irradiating by red light, controlling the light-dark time ratio to be 10-14:14-10, introducing air flow to be 0.4-0.6vvm, culturing for 7-9 days, and controlling the salinity to be 22-26 by adding a sodium chloride aqueous solution with the concentration of 2-3mol/L in the whole culture process;
the isopulegolic algae culture solution is prepared according to the following steps: adding ammonium bicarbonate and brassinolide into the f/2 culture medium;
the concentration of the ammonium bicarbonate is 30-50 mg/L;
the concentration of the brassinolide is 0.02-0.03 mg/L;
the wavelength of the red light is between 720 and 740 nm;
the light intensity of the red light is 45-55 mu mol/(m)2.S）。
2. The process according to claim 1, wherein the seeding density is 500 ten thousand per ml.
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Denomination of invention: A Production Technology for High Density Culture of Isogella spp Effective date of registration: 20221118 Granted publication date: 20220412 Pledgee: Zhejiang Fuyang Rural Commercial Bank Co.,Ltd. Lishan Sub branch Pledgor: HANGZHOU YUSEN AGRICULTURAL TECHNOLOGY DEVELOPMENT Co.,Ltd. Registration number: Y2022980022365 |