Composition for improving aquaculture water quality and preparation method thereof

Abstract

The invention belongs to the technical field of microbial products, and particularly relates to a composition for improving aquaculture water quality and a preparation method thereof. The composition comprises the following raw materials in parts by weight: 10-25 parts of compound bacterial powder, 8-15 parts of humic acid and 3-9 parts of bentonite. A preparation process of the compound bacterium powder comprises the following steps: inoculating a bacillus subtilis bacterium solution and a lactobacillus plantarum bacterium solution into a fermentation culture medium, and carrying out fermentation culture to obtain a fermentation product; and adding maggot powder and zein into the fermentation product, culturing, centrifuging, removing supernatant, and freeze-drying to obtain the compound bacterial powder. The composition for improving the aquaculture water quality can effectively reduce the concentration of ammonia nitrogen, total nitrogen, total phosphorus, nitrite and the like in a water body and improve the ecological environment of an aquaculture water area.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbial products, and in particular relates to a composition for improving aquaculture water quality and a preparation method thereof. Background Art

[0002] Aquaculture is a key pillar of global food production. As the scale of aquaculture expands, water quality deterioration becomes increasingly serious. High-density aquaculture requires the feeding of large quantities of feed. The resulting feces and organic sludge, such as residual brown matter from organisms, also settles at the bottom of the pond, causing the water quality to deteriorate continuously and posing a serious threat to the health and yield of farmed organisms. Furthermore, pollutants such as organic matter, heavy metals, ammonia nitrogen, and nitrite generated during the aquaculture process, if not effectively treated, will pollute the aquaculture environment and affect the quality and yield of aquatic products. The aquaculture industry faces numerous challenges, including severe water pollution, ecological imbalances, and potential safety risks for aquatic products. Traditional water quality improvement methods have numerous limitations, and more efficient and environmentally friendly solutions are urgently needed.

[0003] Microbial agents are microbial bodies screened from natural environments. After cultivation and reproduction, they are made into live bacterial preparations containing a large number of beneficial bacteria. They have been widely used in many fields such as aquaculture, sewage treatment, water and soil improvement. Currently, common microbial preparations for improving water environments have problems such as poor stress resistance, low viable bacterial counts, and short shelf life. They cannot effectively improve the quality of aquaculture water bodies, and the bacteria are easily affected by aquaculture sewage, which limits the promotion and application of microbial preparations in aquaculture water bodies. Bacillus bacteria have the advantages of good stress resistance, rapid growth, strong adaptability, and the ability to produce a rich variety of antibacterial substances. They can reduce the concentrations of ammonia nitrogen, nitrite, sulfide, etc. in water bodies, thereby effectively improving water quality, avoiding the deposition of organic matter in aquaculture ponds, and maintaining a good ecological environment for aquaculture waters. As a probiotic, Lactobacillus plantarum also has many advantages in aquaculture, such as improving water quality, enhancing host health, and promoting growth.

[0004] Based on the above objectives, the present invention provides a composition for improving aquaculture water quality and a preparation method thereof. Summary of the Invention

[0005] The first object of the present invention is to provide a composition for improving aquaculture water quality.

[0006] A second object of the present invention is to provide a method for preparing a composition for improving aquaculture water quality.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is:

[0008] A composition for improving aquaculture water quality, comprising the following raw materials in parts by weight: 10-25 parts of composite bacterial powder, 8-15 parts of humic acid, and 3-9 parts of bentonite; the preparation process of the composite bacterial powder is as follows:

[0009] (1) inoculating a Bacillus subtilis bacterial solution and a Lactobacillus plantarum bacterial solution into a fermentation medium, performing fermentation culture, and obtaining a fermentation product;

[0010] (2) Adding grain insect powder and corn alcohol-soluble protein to the fermentation product, culturing, centrifuging, removing the supernatant, and freeze-drying to obtain composite bacterial powder.

[0011] Furthermore, in step (2), the amount of grain insect powder added is 2-3 times the total weight of the fermentation product; the amount of zein added is 1-2 times the total weight of the fermentation product; and the culture time is 15-24 hours.

[0012] Furthermore, the preparation process of the Bacillus subtilis bacterial solution and the Lactobacillus plantarum bacterial solution in step (1) is: activating and culturing Bacillus subtilis and Lactobacillus plantarum respectively to obtain the Bacillus subtilis bacterial solution and the Lactobacillus plantarum bacterial solution.

[0013] Furthermore, the specific steps of the activation culture are: inoculating Bacillus subtilis into LB culture medium and culturing at 37°C for 24-28 hours to obtain Bacillus subtilis culture liquid; inoculating Lactobacillus plantarum into MRS culture medium and culturing at 37°C for 24-28 hours to obtain Lactobacillus plantarum culture liquid.

[0014] Furthermore, the effective viable bacteria count of the Bacillus subtilis liquid is 1-3×10 8 CFU / mL; the effective viable count of the Lactobacillus plantarum liquid is 1-5×10 8 CFU / mL.

[0015] Furthermore, in step (1), the Bacillus subtilis liquid, the Lactobacillus plantarum liquid and the fermentation medium are inoculated at a volume ratio of 5:(5-9):100; and the fermentation culture time is 20-30 hours.

[0016] Furthermore, the fermentation medium in step (1) comprises the following raw materials in parts by weight: 10-20 parts of glucose, 20-30 parts of bean dregs, 1-5 parts of potassium dihydrogen phosphate, 1-5 parts of calcium carbonate, 5-10 parts of sodium chloride, 1-4 parts of magnesium sulfate, and 40-60 parts of distilled water.

[0017] The preparation method of the above-mentioned composition for improving aquaculture water quality is specifically as follows:

[0018] The composite bacterial powder, humic acid and bentonite are weighed according to the stated parts by weight, and stirred evenly at room temperature to obtain the product.

[0019] Compared with the prior art, the beneficial effects of the present invention are mainly:

[0020] The present invention can effectively improve the stability of the composite bacterial powder by adding grain insect powder and corn alcohol-soluble protein to the fermentation product, so that the microorganisms in the composite bacterial powder maintain good activity and promote bacterial proliferation. Combined with humic acid and bentonite, it can significantly improve water quality, effectively reduce the concentrations of ammonia nitrogen, total nitrogen, total phosphorus, nitrite, etc. in the water body, and improve the ecological environment of aquaculture waters. DETAILED DESCRIPTION

[0021] The technical solution of the present invention will be further described below in conjunction with specific embodiments. However, it should be understood by those skilled in the art that the following examples are only intended to illustrate the present invention and should not be construed as limiting the present invention. Specific conditions not specified in the examples are to be followed according to conventional conditions or the conditions recommended by the manufacturer. All reagents or instruments used, unless otherwise specified, are conventional products obtained from commercial channels.

[0022] The preparation method of the grain insect powder used in the embodiment of the present invention is:

[0023] The grain worms were dried at 80° C. for 4 h and then crushed in a grinder for 30 min to obtain the grain worm powder used in this example.

[0024] The Lactobacillus plantarum of the invention has a preservation number of CGMCC1.557; the Bacillus subtilis has a preservation number of CGMCC1.921.

[0025] Example 1

[0026] A composition for improving aquaculture water quality comprises the following raw materials in parts by weight: 17 parts of composite bacterial powder, 12 parts of humic acid, and 6 parts of bentonite.

[0027] The preparation method of the composite bacterial powder specifically comprises the following steps:

[0028] (1) Bacillus subtilis was inoculated into 2 mL of LB medium and cultured at 37°C for 26 h to obtain a Bacillus subtilis culture solution; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 26 h to obtain a Lactobacillus plantarum culture solution; the effective viable cell count of the Bacillus subtilis culture solution was 2×10 8 CFU / mL; the effective viable count of the Lactobacillus plantarum liquid is 3×10 8 CFU / mL.

[0029] (2) To 50 parts of distilled water, 15 parts of glucose, 25 parts of soybean dregs, 3 parts of potassium dihydrogen phosphate, 3 parts of calcium carbonate, 7 parts of sodium chloride, and 2 parts of magnesium sulfate were added by weight to prepare a fermentation medium.

[0030] (3) Inoculate the Bacillus subtilis solution, Lactobacillus plantarum solution and fermentation medium at a volume ratio of 5:7:100, and perform fermentation culture for 25 hours to obtain a fermentation product.

[0031] (4) Adding grain insect powder and zein to the fermentation product of step (3), wherein the amount of grain insect powder added is 3 times the total weight of the fermentation product; the amount of zein added is 2 times the total weight of the fermentation product; after culturing for 19 hours, centrifugation is performed, the supernatant is removed, and freeze-drying is performed to obtain a composite bacterial powder.

[0032] This embodiment also provides a method for preparing a composition for improving aquaculture water quality, which specifically comprises the following steps:

[0033] The composite bacterial powder, humic acid and bentonite are weighed according to the stated parts by weight, and stirred evenly at room temperature to obtain the product.

[0034] Example 2

[0035] A composition for improving aquaculture water quality comprises the following raw materials in parts by weight: 10 parts of composite bacterial powder, 8 parts of humic acid, and 3 parts of bentonite.

[0036] The preparation method of the composite bacterial powder specifically comprises the following steps:

[0037] (1) Bacillus subtilis was inoculated into 2 mL of LB medium and cultured at 37°C for 24 h to obtain a Bacillus subtilis culture solution; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 24 h to obtain a Lactobacillus plantarum culture solution; the effective viable cell count of the Bacillus subtilis culture solution was 1×10 8 CFU / mL; the effective viable count of the Lactobacillus plantarum liquid is 1×10 8 CFU / mL.

[0038] (2) To 40 parts of distilled water, 10 parts of glucose, 20 parts of soybean dregs, 1 part of potassium dihydrogen phosphate, 1 part of calcium carbonate, 5 parts of sodium chloride, and 1 part of magnesium sulfate were added, by weight, to prepare a fermentation medium.

[0039] (3) Inoculate the Bacillus subtilis solution, the Lactobacillus plantarum solution and the fermentation medium at a volume ratio of 5:5:100, and perform fermentation culture for 20 hours to obtain a fermentation product.

[0040] (4) Adding grain insect powder and zein to the fermentation product of step (3), wherein the amount of grain insect powder added is 2 times the total weight of the fermentation product; the amount of zein added is 1 times the total weight of the fermentation product; after culturing for 15 hours, centrifuging, removing the supernatant, and freeze-drying to obtain a composite bacterial powder.

[0041] This embodiment also provides a method for preparing a composition for improving aquaculture water quality, which specifically comprises the following steps:

[0042] The composite bacterial powder, humic acid and bentonite are weighed according to the stated parts by weight, and stirred evenly at room temperature to obtain the product.

[0043] Example 3

[0044] A composition for improving aquaculture water quality comprises the following raw materials in parts by weight: 25 parts of composite bacterial powder, 15 parts of humic acid, and 9 parts of bentonite.

[0045] The preparation method of the composite bacterial powder specifically comprises the following steps:

[0046] (1) Bacillus subtilis was inoculated into 2 mL of LB medium and cultured at 37°C for 28 h to obtain a Bacillus subtilis culture solution; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 28 h to obtain a Lactobacillus plantarum culture solution; the effective viable cell count of the Bacillus subtilis culture solution was 3×10 8 CFU / mL; the effective viable count of the Lactobacillus plantarum liquid is 5×10 8 CFU / mL.

[0047] (2) To 60 parts of distilled water, 20 parts of glucose, 30 parts of soybean dregs, 5 parts of potassium dihydrogen phosphate, 5 parts of calcium carbonate, 10 parts of sodium chloride, and 4 parts of magnesium sulfate were added by weight to prepare a fermentation medium.

[0048] (3) Inoculate the Bacillus subtilis solution, the Lactobacillus plantarum solution and the fermentation medium at a volume ratio of 5:9:100, and perform fermentation culture for 30 hours to obtain a fermentation product.

[0049] (4) Adding grain insect powder and zein to the fermentation product of step (3), wherein the amount of grain insect powder added is 3 times the total weight of the fermentation product; the amount of zein added is 2 times the total weight of the fermentation product; after culturing for 24 hours, centrifuging, removing the supernatant, and freeze-drying to obtain a composite bacterial powder.

[0050] This embodiment also provides a method for preparing a composition for improving aquaculture water quality, which specifically comprises the following steps:

[0051] The composite bacterial powder, humic acid and bentonite are weighed according to the stated parts by weight, and stirred evenly at room temperature to obtain the product.

[0052] Comparative Example 1

[0053] The difference between Comparative Example 1 and Example 1 is that the grain and insect powder is omitted from the composite fungus powder prepared in step (4).

[0054] Comparative Example 2

[0055] The difference between Comparative Example 2 and Example 1 is that zein is omitted from the composite bacterial powder component prepared in step (4).

[0056] Comparative Example 3

[0057] The difference between Comparative Example 3 and Example 1 is that the grain insect powder is omitted from the composite bacterial powder component prepared in step (4), and the amount of zein is adjusted to the sum of the two. The rest is the same as Example 1.

[0058] Comparative Example 4

[0059] The difference between Comparative Example 4 and Example 1 is that zein is omitted from the composite bacterial powder component prepared in step (4), and the amount of grain insect powder is adjusted to the sum of the two. The rest is the same as Example 1.

[0060] Test Example 1

[0061] The following is an effect test of the composition for improving aquaculture water quality obtained in Examples 1-3 and Comparative Examples 1-4:

[0062] 14L of aquaculture water was taken and divided into Example 1, Example 2, Example 3, Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4, with 2L in each group. 2g of the composition for improving aquaculture water quality prepared in the corresponding groups of Examples 1-3 and Comparative Examples 1-4 was added respectively, once every three days. After three weeks of continuous feeding, the COD and NH4 levels of the water after adding the composition for improving aquaculture water quality were tested. + -N, total nitrogen, total phosphorus, NO2 - The contents of various components in the water before the test are shown in Table 1, and the contents of various components in the water after adding the composition for improving aquaculture water quality are shown in Table 2.

[0063] Table 1 Contents of various components in water before the test

[0064]

[0065] Table 2 Contents of various components in water after adding the composition for improving aquaculture water quality

[0066]

[0067] The results are shown in Table 1 and Table 2, which are the contents of various components in the water before the test and after adding the composition for improving aquaculture water quality. As can be seen from the table, after continuous feeding for 3 weeks, the composition for improving aquaculture water quality prepared by Examples 1-3 of the present invention can significantly reduce the COD content and NH4 + -N content, total nitrogen content, total phosphorus content, NO2 - -N content. It shows that the bentonite, humic acid and composite bacterial powder in the composition for improving aquaculture water quality prepared by the present invention can be used together to effectively reduce the concentrations of ammonia nitrogen, total nitrogen, total phosphorus and nitrite in the water body, thereby improving the ecological environment of aquaculture waters.

[0068] This is because bentonite has a strong adsorption capacity and can directly absorb toxic substances such as ammonia nitrogen and nitrite. Humic acid can improve aquaculture water quality and promote the healthy growth of aquatic products, while also reducing the use of chemicals and enhancing the water's self-purification capacity. Adding grain insect powder and zein to the fermentation product effectively improves the stability of the composite bacterial powder, maintains the active microorganisms within it, promotes bacterial proliferation, and reduces the concentrations of ammonia nitrogen, total nitrogen, total phosphorus, and nitrite in the water.

[0069] Further analysis revealed that compared to Example 1, the composite powder prepared in Comparative Example 1 omitted grain insect powder; the composite powder prepared in Comparative Example 2 omitted zein; the composite powder prepared in Comparative Example 3 omitted grain insect powder and adjusted the amount of zein to the sum of the two; and the composite powder prepared in Comparative Example 4 omitted zein and adjusted the amount of grain insect powder to the sum of the two. The effectiveness of all four groups in improving the ecological environment of aquaculture waters was significantly reduced. This suggests that the grain insect powder and zein in the composite powder have a synergistic effect, and their combined use effectively improves the stability of the composite powder, maintains good microbial activity within the composite powder, promotes bacterial proliferation, effectively reduces concentrations of ammonia nitrogen, total nitrogen, total phosphorus, nitrite, and other substances in the water, and improves the ecological environment of aquaculture waters.

[0070] Finally, it should be noted that the above embodiments are intended only to illustrate the technical solutions of the present invention and are not intended to limit the same. The basic principles and main features of the present invention have been described above using specific implementation schemes. Modifications or substitutions may be made based on the present invention, but such modifications or substitutions do not detract from the essence of the corresponding technical solutions from the scope of protection claimed by the present invention.

Claims

1. A composition for improving aquaculture water quality, characterized in that: The composition comprises the following raw materials in parts by weight: 10-25 parts of composite bacterial powder, 8-15 parts of humic acid, and 3-9 parts of bentonite. The preparation process of the composite bacterial powder is as follows: (1) inoculating a Bacillus subtilis bacterial solution and a Lactobacillus plantarum bacterial solution into a fermentation medium, performing fermentation culture, and obtaining a fermentation product; (2) Adding grain insect powder and corn alcohol-soluble protein to the fermentation product, culturing, centrifuging, removing the supernatant, and freeze-drying to obtain composite bacterial powder.

2. A composition for improving aquaculture water quality according to claim 1, characterized in that: In step (2), the amount of grain insect powder added is 2-3 times the total weight of the fermentation product; the amount of zein added is 1-2 times the total weight of the fermentation product; and the culture time is 15-24 hours.

3. A composition for improving aquaculture water quality according to claim 1, characterized in that: The preparation process of the Bacillus subtilis bacterial solution and the Lactobacillus plantarum bacterial solution in the step (1) is as follows: Bacillus subtilis and Lactobacillus plantarum are activated and cultured respectively to obtain the Bacillus subtilis bacterial solution and the Lactobacillus plantarum bacterial solution.

4. A composition for improving aquaculture water quality according to claim 3, characterized in that: The specific steps of the activation culture are: inoculating Bacillus subtilis into LB culture medium, culturing at 37°C for 24-28 hours to obtain Bacillus subtilis bacterial liquid; inoculating Lactobacillus plantarum into MRS culture medium, culturing at 37°C for 24-28 hours to obtain Lactobacillus plantarum bacterial liquid.

5. A composition for improving aquaculture water quality according to claim 3, characterized in that: The effective viable bacteria count of the Bacillus subtilis liquid is 1-3×10 8 CFU / mL; the effective viable count of the Lactobacillus plantarum liquid is 1-5×10 8 CFU / mL.

6. A composition for improving aquaculture water quality according to claim 1, characterized in that: In the step (1), the Bacillus subtilis liquid, the Lactobacillus plantarum liquid and the fermentation medium are inoculated at a volume ratio of 5:(5-9):100; and the fermentation culture time is 20-30 hours.

7. A composition for improving aquaculture water quality according to claim 1, characterized in that: The fermentation medium in step (1) comprises the following raw materials in parts by weight: 10-20 parts of glucose, 20-30 parts of bean dregs, 1-5 parts of potassium dihydrogen phosphate, 1-5 parts of calcium carbonate, 5-10 parts of sodium chloride, 1-4 parts of magnesium sulfate, and 40-60 parts of distilled water.

8. The method for preparing the composition for improving aquaculture water quality according to any one of claims 1 to 7, characterized in that: The specific preparation method is as follows: The composite bacterial powder, humic acid and bentonite are weighed according to the stated parts by weight, and stirred evenly at room temperature to obtain the product.

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