Fungus bag, fungus bag preparation method and green alga cultivation method

Abstract

The invention discloses a fungus bag, a fungus bag preparation method and a green alga cultivation method, and belongs to the technical field of green alga cultivation. The fungus bag is prepared from the following raw materials in parts by weight: 200 to 400 parts of plant base stock, 10 to 20 parts of brown sugar, 10 to 20 parts of urea, 4 to 8 parts of calcium carbonate, 2 to 4 parts of ferrous sulfate, 2 to 4 parts of ammonium sulfate, 1.8 to 4 parts of monopotassium phosphate, 0.3 to 1 part of zinc sulfate, 10 to 15 parts of bacillus velezensis liquid, 10 to 15 parts of bacillus marinensis liquid and the balance of sterilized seawater. The fungus bag can effectively solve the problems that in the prior art, culture water is prone to yellowing, the activity of strains is reduced, and diseases and insect pests are prone to occurring.

Description

Technical Field

[0001] The present invention belongs to the technical field of green algae cultivation, and particularly relates to a bacteria bag, a method for preparing the bacteria bag, and a method for culturing green algae. Background Art

[0002] Algae of the phylum Chlorophyta are widely used in aquaculture as biological baits and biological water quality improvers. They have the characteristic of fast growth rate. Under photosynthesis, they consume metabolic wastes such as ammonia nitrogen, phosphorus, and inorganic carbon in the aquaculture water body and produce oxygen, thus playing the role of carbon fixation, increasing the dissolved oxygen in the aquaculture water body, and maintaining the stability of the aquaculture water environment. At the same time, they are rich in nutrients, especially Chlorella and Nannochloropsis, which are rich in unsaturated fatty acids and other nutrient elements. Among them, the content of EPA accounts for about 30% of the total fatty acids. EPA has an anti-inflammatory effect, can significantly improve the survival rate of fry, and reduce the aquaculture cost.

[0003] Currently, the main problems in the process of industrialized seawater cultivation of green algae are that the algae liquid is prone to turn yellow, the vitality of the algal species declines, and pathogenic organisms break out under continuous culture conditions, resulting in the inability to continuously produce. Especially when the disinfection of the water used for culturing green algae is not standardized and thorough, the aquaculture environment is relatively poor, and the biological pollution is serious, it often leads to the failure of algae cultivation, causing losses and at the same time the residual nutrients also aggravate the water pollution. Currently, for the above problems, except for strengthening the disinfection of the water body, there is no better solution. Summary of the Invention

[0004] In view of the above deficiencies in the prior art, the present invention provides a bacteria bag, a method for preparing the bacteria bag, and a method for culturing green algae. The bacteria bag can effectively solve the problems of easy yellowing of the aquaculture water body, decline in the vitality of the bacterial species, and easy occurrence of pests and diseases existing in the prior art.

[0005] To achieve the above object, the technical solution adopted by the present invention to solve its technical problems is:

[0006] A bacteria bag, comprising the following raw materials in parts by weight: 200 - 400 parts of plant base material, 10 - 20 parts of brown sugar, 10 - 20 parts of urea, 4 - 8 parts of calcium carbonate, 2 - 4 parts of ferrous sulfate, 2 - 4 parts of ammonium sulfate, 1.8 - 4 parts of potassium dihydrogen phosphate, 0.3 - 1 part of zinc sulfate, 10 - 15 parts of Bacillus velezensis liquid, 10 - 15 parts of Bacillus haynesii liquid, and the rest is sterilized seawater.

[0007] Further, it also includes 1 - 3 parts of yeast extract or yeast powder.

[0008] Further, the plant base material includes at least one of peanut meal, dried bean dregs, and soybean meal.

[0009] Further, the Bacillus velezensis liquid and the Bacillus haynesii liquid are prepared by the following method:

[0010] (1) Activate the freeze-dried powder of Bacillus velezensis and the freeze-dried powder of Bacillus haynesii with fresh water medium respectively, and then carry out primary amplification culture respectively to obtain the fresh water bacterial liquid of Bacillus velezensis and the fresh water bacterial liquid of Bacillus haynesii.

[0011] (2) Add the fresh water bacterial liquid of Bacillus velezensis and the fresh water bacterial liquid of Bacillus haynesii into the sea water medium respectively for secondary amplification culture to obtain.

[0012] Further, the mass-volume ratio of the freeze-dried powder of Bacillus velezensis and the freeze-dried powder of Bacillus haynesii to the fresh water medium is 0.1-1g:500ml.

[0013] Further, the fresh water medium is fresh water beef extract peptone medium or fresh water potato medium; the sea water medium is sea water beef extract peptone medium or sea water potato medium.

[0014] Further, in step (1), add yeast extract or yeast powder to the medium during primary amplification culture until a large amount of gas is generated.

[0015] Further, in step (2), the volume ratio of the fresh water bacterial liquid of Bacillus velezensis and the fresh water bacterial liquid of Bacillus haynesii to the sea water medium is both 1:180-220; both are amplified until a large amount of gas is generated.

[0016] The preparation method of the above-mentioned bacteria bag includes the following steps: Mix the raw materials except sterilized sea water evenly to obtain a mixture, add an appropriate amount of sterilized sea water to the mixture, stir and mix evenly until it becomes a dry mud state, and then put it into a polyethylene mesh bag with 100-160 meshes to obtain.

[0017] A method for culturing green algae includes the following steps: Inject the green algae into the algae pond, and then evenly place the bacteria bags into the algae pond according to the amount of 1 bacteria bag per 9-11 cubic meters.

[0018] Further, the bacteria bags are placed into the algae pond 3-5 days after the green algae are injected into the algae pond.

[0019] The beneficial effects produced by the present invention are as follows:

[0020] In the present invention, by utilizing the strain characteristics of Bacillus haynesii and Bacillus velezensis, the two are made into a slow-release bacteria bag, which is added with urea and trace elements. The urea, trace elements, etc. in the bacteria bag are slowly released, reducing the peak concentration of nutrients in the algae pond and reducing the toxic effects caused by excessive nutrient concentration.

[0021] Bacillus velezensis is a type of spore-forming Gram-positive bacteria that can secrete a variety of bioactive substances, including enzymes, antibacterial proteins, lipopeptide antibiotics, polyketide antibiotics, plant hormones, etc., and can convert ammonia nitrogen into nitrate nitrogen through nitrification and denitrification; Bacillus haynesii is a Gram-positive, facultative anaerobic, spore-forming rod-shaped bacterium that can secrete xylanase and keratinase, and the secreted products have a strong effect on killing protozoa. In this application, small peptides, plant hormones, nitrate nitrogen produced by nitrification, carbon dioxide produced by respiration, etc. generated during the growth of the bacteria are used to promote the growth of green algae; finally, the active substances secreted by the bacteria are used to inhibit the growth of pathogenic organisms, obtaining better cultivation conditions, effectively preserving the nutrients of the algae themselves, improving the carbon fixation ability of the algae, and effectively increasing the dry matter content of the algal solution. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a statistical chart of the culture days and the biomass of harmful organisms;

[0023] Figure 2 It is a statistical chart of the insecticidal effect of the bacterial culture solution. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0024] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention, that is, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments.

[0025] Therefore, the following detailed description of the provided embodiments of the present invention is not intended to limit the scope of the claimed present invention, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative efforts fall within the scope of protection of the present invention.

[0026] It should be noted that relational terms such as "first" and "second" are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprising", "including" or any other variant thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements not only includes those elements, but also includes other elements not expressly listed, or also includes elements inherent to such process, method, article or device. Without further limitation, an element defined by the statement "including one..." does not exclude the presence of additional identical elements in the process, method, article or device including the element.

[0027] The features and performance of the present invention will be further described in detail below in conjunction with the embodiments and the accompanying drawings.

[0028] Example 1

[0029] A fungus bag comprises raw materials in the following parts by weight: 200 parts of peanut meal, 10 parts of brown sugar, 10 parts of urea, 4 parts of calcium carbonate, 2 parts of ferrous sulfate, 2 parts of ammonium sulfate, 1.8 parts of potassium dihydrogen phosphate, 0.3 part of zinc sulfate, 10 parts of Bacillus velezensis liquid, 10 parts of Bacillus haynesii liquid, 3 parts of yeast extract, and the rest is sterilized seawater.

[0030] The Bacillus velezensis liquid and the Bacillus haynesii liquid are prepared by the following method:

[0031] (1) 0.2 g of Bacillus velezensis freeze-dried powder and 0.2 g of Bacillus haynesii freeze-dried powder are respectively activated with 500 ml of fresh water beef extract peptone medium for 3 days. At this time, obvious gas production phenomenon and surface bacterial film can be seen in Bacillus velezensis, and a large number of rod-shaped bacteria with a length of about 6 - 10 μm can be seen under microscopic examination; there is no gas production phenomenon in Bacillus haynesii, there is a relatively thin bacterial film on the surface, and a large number of curved bacteria with a length of about 0.5 - 2 μm can be obtained under microscopic examination; then 3 g of yeast extract is respectively added to the medium for primary expansion culture until a large amount of gas is produced, and the fresh water bacterial liquid of Bacillus velezensis and the fresh water bacterial liquid of Bacillus haynesii are obtained;

[0032] (2) 500 ml of the fresh water bacterial liquid of Bacillus velezensis and 500 ml of the fresh water bacterial liquid of Bacillus haynesii are respectively added into 90 L of seawater beef extract peptone medium for secondary expansion culture until a large amount of gas is produced, and then obtained.

[0033] The preparation method of the above-mentioned fungus bag comprises the following steps: mixing the raw materials except the sterilized seawater evenly to obtain a mixture, adding an appropriate amount of sterilized seawater to the mixture, stirring and mixing evenly until it becomes dry mud-like, and then filling it into a polyethylene 100-mesh polyethylene mesh bag to obtain.

[0034] A green algae cultivation method comprises the following steps: injecting green algae into an algae pond, and after 3 days, evenly placing the fungus bags into the algae pond according to the amount of 1 fungus bag per 9 cubic meters.

[0035] Example 2

[0036] A fungus bag comprises raw materials in the following parts by weight: 400 parts of dried bean dregs, 20 parts of brown sugar, 20 parts of urea, 8 parts of calcium carbonate, 4 parts of ferrous sulfate, 4 parts of ammonium sulfate, 4 parts of potassium dihydrogen phosphate, 1 part of zinc sulfate, 15 parts of Bacillus velezensis liquid, 15 parts of Bacillus haynesii liquid, 3 parts of yeast extract, and the rest is sterilized seawater.

[0037] The Bacillus velezensis liquid and the Bacillus haynesii liquid are prepared by the following method:

[0038] (1) 1 g of freeze-dried powder of Bacillus velezensis and 1 g of freeze-dried powder of Bacillus haynesii are respectively activated with 500 ml of fresh water beef extract peptone medium for 3 days. At this time, obvious gas production phenomenon and surface bacterial film can be seen in Bacillus velezensis, and a large number of rod-shaped bacteria with a length of about 6-10 μm can be seen under microscopic examination; there is no gas production phenomenon in Bacillus haynesii, and there is a relatively thin bacterial film on the surface, and a large number of curved bacteria with a length of about 0.5-2 μm can be obtained under microscopic examination; then 1 g of yeast powder is respectively added to the medium for primary expansion culture until a large amount of gas is produced, and the fresh water bacterial liquid of Bacillus velezensis and the fresh water bacterial liquid of Bacillus haynesii are prepared;

[0039] (2) 500 ml of the fresh water bacterial liquid of Bacillus velezensis and 500 ml of the fresh water bacterial liquid of Bacillus haynesii are respectively added into 110 L of seawater beef extract peptone medium for secondary expansion culture until a large amount of gas is produced, and then obtained.

[0040] The preparation method of the above-mentioned bacterial bag includes the following steps: mixing the raw materials except sterilized seawater evenly to obtain a mixture, adding an appropriate amount of sterilized seawater to the mixture, stirring and mixing evenly until it becomes dry mud, and then filling it into a polyethylene mesh bag with 160 meshes to obtain.

[0041] A method for culturing green algae includes the following steps: injecting green algae into an algae pond, and after 3 days, evenly placing the bacterial bags into the algae pond according to the amount of 1 bacterial bag per 11 cubic meters.

[0042] Example 3

[0043] A bacterial bag includes the following raw materials in parts by weight: 300 parts of soybean meal, 15 parts of brown sugar, 15 parts of urea, 6 parts of calcium carbonate, 3 parts of ferrous sulfate, 3 parts of ammonium sulfate, 3 parts of potassium dihydrogen phosphate, 2 parts of zinc sulfate, 13 parts of Bacillus velezensis liquid, 13 parts of Bacillus haynesii liquid, 3 parts of yeast extract, and the rest is sterilized seawater.

[0044] The Bacillus velezensis liquid and the Bacillus haynesii liquid are prepared by the following method:

[0045] (1) 0.7 g of freeze-dried powder of Bacillus velezensis and 0.7 g of freeze-dried powder of Bacillus haynesii were respectively activated in 500 ml of fresh water beef extract peptone medium for 3 days. At this time, obvious gas production phenomenon and surface bacterial film could be seen in Bacillus velezensis, and a large number of rod-shaped bacteria with a length of about 6 - 10 μm could be seen under microscopic examination; there was no gas production phenomenon in Bacillus haynesii, and there was a relatively thin bacterial film on the surface, and a large number of curved bacteria with a length of about 0.5 - 2 μm could be obtained under microscopic examination. Then, 3 g of yeast extract was added to the medium respectively for primary expansion culture until a large amount of gas was produced, and the fresh water bacterial solution of Bacillus velezensis and the fresh water bacterial solution of Bacillus haynesii were prepared.

[0046] (2) 500 ml of the fresh water bacterial solution of Bacillus velezensis and 500 ml of the fresh water bacterial solution of Bacillus haynesii were respectively added into 100 L of seawater beef extract peptone medium for secondary expansion culture until a large amount of gas was produced.

[0047] The preparation method of the above-mentioned bacterial bag includes the following steps: Mix the raw materials except sterilized seawater evenly to obtain a mixture, add an appropriate amount of sterilized seawater to the mixture, stir and mix evenly until it becomes dry mud, and then put it into a polyethylene mesh bag with 120 meshes to obtain the bacterial bag.

[0048] A method for culturing green algae includes the following steps: Inject green algae into the algae pond. After 3 days, put the bacterial bags into the algae pond evenly according to the amount of 1 bacterial bag per 10 cubic meters.

[0049] Example 4

[0050] A bacterial bag includes the following raw materials in parts by weight: 350 parts of peanut meal, 16 parts of brown sugar, 17 parts of urea, 5 parts of calcium carbonate, 3 parts of ferrous sulfate, 4 parts of ammonium sulfate, 3.5 parts of potassium dihydrogen phosphate, 0.8 parts of zinc sulfate, 14 parts of Bacillus velezensis liquid, 14 parts of Bacillus haynesii liquid, 3 parts of yeast extract, and the rest is sterilized seawater.

[0051] The Bacillus velezensis liquid and the Bacillus haynesii liquid are prepared by the following method:

[0052] (1) 0.3 g of freeze-dried powder of Bacillus velezensis and 0.3 g of freeze-dried powder of Bacillus haynesii were respectively activated in 500 ml of fresh water beef extract peptone medium for 3 days. At this time, obvious gas production phenomenon and surface bacterial film could be seen in Bacillus velezensis, and a large number of rod-shaped bacteria with a length of about 6 - 10 μm could be seen under microscopic examination; there was no gas production phenomenon in Bacillus haynesii, and there was a relatively thin bacterial film on the surface, and a large number of curved bacteria with a length of about 0.5 - 2 μm could be obtained under microscopic examination. Then, 3 g of yeast extract was added to the medium respectively for primary expansion culture until a large amount of gas was produced, and the fresh water bacterial solution of Bacillus velezensis and the fresh water bacterial solution of Bacillus haynesii were prepared.

[0053] (2) Add 500 ml of the fresh water Bacillus velezensis bacterial liquid and 500 ml of the fresh water Bacillus haynesii bacterial liquid into 105 L of the seawater beef extract peptone medium respectively for secondary expansion culture until a large amount of gas is generated, thus obtaining.

[0054] The preparation method of the above-mentioned bacterial bag includes the following steps: Mix the raw materials except the sterilized seawater evenly to obtain a mixture, add an appropriate amount of sterilized seawater to the mixture, stir and mix evenly until it becomes dry mud-like, and then load it into a polyethylene mesh bag with 140 meshes to obtain.

[0055] A method for culturing green algae includes the following steps: Inject the green algae into the algae pond. After 3 days, evenly place the bacterial bags into the algae pond according to the amount of 1 bacterial bag per 11 cubic meters.

[0056] Test Example

[0057] Take natural seawater, filter it through a sand filter pond, and then filter it successively through a 500-mesh polyethylene mesh bag and a polyester polypropylene filter bag (PP filter bag) with a filtration accuracy of 25 μm, and then introduce it into 6 algae cultivation ponds with a size of 4.8 m * 4.8 m and a water depth of 0.8 m, with nutrient salts: urea 108 mg / L, KH 2 PO 4 12 mg / L, NaHCO 3 500 mg / L for algae cultivation.

[0058] Take the day of inoculating the algal liquid as day 0. Randomly sample and detect and record the amount of harmful organisms in each pond at 10:00 every morning. At 8:00 in the morning on the 3rd day after inoculating the algal liquid, put the bacterial bags prepared in Example 3 into 3 randomly selected ponds as experimental ponds, and the remaining 3 algae cultivation ponds without putting the bacterial bags as control ponds.

[0059] Take the number of harmful organisms exceeding 100 individuals / ml and the cultivation time of 35 days as the standard for the end of the experiment, and stop recording the relevant data. Record and statistically organize the average value of the experimental data. The specific results are shown in Figure 1 .

[0060] Figure 1 The results show that in the control ponds, starting from the 4th day, the amount of harmful organisms shows an upward trend, and the amount of harmful organisms exceeds 100 individuals / ml on the 17th day. While in the experimental ponds with the added bacterial bags, the content of harmful organisms has been relatively low, proving that the bacterial bags in the present invention can effectively inhibit the number of harmful organisms in the algae pond, delay the time of problems such as the algal liquid turning yellow and the vitality decreasing from 12 - 18 days to more than 30 days, and improve the growth rate of algae on the basis of no harmful organisms.

[0061] Take the bacterial liquid culture of the bacteria in the inoculated bacterial bag for 8 days, and dilute it to 2 0 、2 1 、2 2 、23 , 2 4 , 2 5 , 2 6 , 2 7 , 2 8 , 2 9 , 2 10 times. Using rotifers as the bioindicator of toxicity, more than 20 live rotifers were placed into each concentration of the diluted solution and timing was started. The standard for rotifer death was that all rotifer bodies stopped moving and their mouthparts were immobile. The half-death time and full-death time of the rotifers were recorded. The same experiment was repeated 3 times to calculate the average value. The specific results are shown in Figure 2 .

[0062] Figure 2 The results showed that the culture solutions of Bacillus haynesii and Bacillus velezensis had good killing effects on rotifers.

[0063] Take 5 L each of the algal solution cultured without the bacterial bag (control group) and the algal solution cultured with the bacterial bag (experimental group). Use a suction filtration device equipped with a 1-μm pore size polypropylene microporous filter membrane to filter and separate the green algae. Place the green algae in an oven at 105 °C and dry to a constant weight, and compare the dry matter amounts of the two.

[0064] The results showed that the average dry matter of the control group was 21.82 g, and the average dry matter of the experimental group was 27.22 g, that is, the dry matter amount of the algal solution cultured with the bacterial bag was more than 20% higher than that of the algal solution cultured without the bacterial bag.

Claims

1. A mushroom bag, characterized in that: The invention comprises the following raw materials in parts by weight: 200-400 parts of plant base material, 10-20 parts of brown sugar, 10-20 parts of urea, 4-8 parts of calcium carbonate, 2-4 parts of ferrous sulfate, 2-4 parts of ammonium sulfate, 1.8-4 parts of potassium dihydrogen phosphate, 0.3-1 part of zinc sulfate, 10-15 parts of Bacillus veleus liquid, 10-15 parts of Bacillus henesii liquid, and the rest is sterilized seawater.

2. The mushroom bag according to claim 1, characterized in that: It also includes 1-3 parts of yeast extract or yeast powder.

3. The mushroom bag according to claim 1, characterized in that: The plant-based material comprises at least one of peanut meal, dry bean dregs and soybean meal.

4. The mushroom bag according to claim 1, characterized in that: The Bacillus Velez liquid and the Bacillus Heines liquid are prepared by the following method: (1) activating the freeze-dried powder of Bacillus Velez and the freeze-dried powder of Bacillus henesii respectively with fresh water culture medium, and then performing primary expansion culture respectively to obtain fresh water bacterial liquid of Bacillus Velez and fresh water bacterial liquid of Bacillus henesii; (2) Adding freshwater bacterial liquid of Bacillus velez and freshwater bacterial liquid of Bacillus heines into seawater culture medium respectively for secondary expansion culture to obtain the product.

5. The mushroom bag according to claim 4, characterized in that: The freshwater culture medium is a freshwater beef extract peptone culture medium or a freshwater potato culture medium; the seawater culture medium is a seawater beef extract peptone culture medium or a seawater potato culture medium.

6. The mushroom bag according to claim 4, characterized in that: In step (1), during the primary expansion, yeast extract or yeast powder is added to the culture medium and the culture is expanded until a large amount of gas is generated.

7. The mushroom bag according to claim 4, characterized in that: In step (2), the volume ratio of the freshwater bacterial liquid of Bacillus velez and the freshwater bacterial liquid of Bacillus heines to the seawater culture medium is 1:180-220; the culture is expanded in the seawater culture medium until a large amount of gas is generated.

8. The method for preparing a mushroom bag according to any one of claims 1 to 7, comprising the following steps: The raw materials except the sterilized seawater are uniformly mixed to obtain a mixture, an appropriate amount of sterilized seawater is added to the mixture, and the mixture is stirred and mixed until it is in a dry mud state, and then loaded into a polyethylene mesh bag with a mesh size of 100-160 to obtain the mixture.

9. A method for cultivating green algae, characterized in that: The method comprises the following steps: injecting green algae into the algae pond, and then evenly placing the fungus bags in the algae pond at a rate of one fungus bag per 9-11 cubic meters.

10. The breeding method according to claim 9, characterized in that: The fungus bag is placed in the algae pond 3-5 days after the green algae are injected into the algae pond.

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