A composition for improving aquaculture water quality and its preparation method

Abstract

This invention belongs to the field of microbial product technology, specifically relating to a composition for improving aquaculture water quality and its preparation method. 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. The preparation process of the compound bacterial powder is as follows: Bacillus subtilis and Lactobacillus plantarum bacterial solutions are inoculated into a fermentation medium and fermented to obtain a fermentation product; grain insect powder and zein are added to the fermentation product, followed by culturing, centrifugation, removal of the supernatant, and freeze-drying to obtain the compound bacterial powder. The composition for improving aquaculture water quality prepared by this invention can effectively reduce the concentrations of ammonia nitrogen, total nitrogen, total phosphorus, and nitrite in the water, thereby improving the ecological environment of aquaculture waters.

Description

Technical Field

[0001] This invention belongs to the field of microbial product technology, specifically relating to a composition for improving aquaculture water quality and its preparation method. Background Technology

[0002] Traditional water quality improvement methods have many limitations, and there is an urgent need for more efficient and environmentally friendly solutions.

[0003] Microbial agents are live bacterial preparations containing a large number of beneficial bacteria, produced by culturing and propagating microbial cells screened from the natural environment. They have been widely used in aquaculture, wastewater treatment, soil improvement, and many other fields. Currently, common microbial preparations for improving aquatic environments suffer from poor stress resistance, low viable cell counts, and short shelf life, failing to effectively improve the quality of aquaculture water. Furthermore, the bacterial strains are easily affected by aquaculture wastewater, thus limiting the widespread application of microbial preparations in aquaculture water. Bacillus bacteria, on the other hand, possess advantages such as good stress resistance, rapid growth, strong adaptability, and the ability to produce a rich variety of antibacterial substances. They reduce the concentrations of ammonia nitrogen, nitrite, and sulfides in the water, effectively improving water quality, preventing the deposition of organic matter in aquaculture ponds, and maintaining a healthy aquatic ecosystem. Lactobacillus plantarum, as a probiotic, also offers various advantages in aquaculture, including improving water quality, enhancing host health, and promoting growth.

[0004] To achieve the above objectives, the present invention provides a composition for improving aquaculture water quality and a method for preparing the same. Summary of the Invention

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

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

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0008] A composition for improving aquaculture water quality, the composition comprising 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; the preparation process of the compound bacterial powder is as follows:

[0009] (1) Inoculate Bacillus subtilis culture and Lactobacillus plantarum culture into fermentation medium and carry out fermentation culture to obtain fermentation products;

[0010] (2) Add grain insect powder and zein to the fermentation product, culture, centrifuge, remove the supernatant, freeze dry to obtain compound 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 corn gliadin added is 1-2 times the total weight of the fermentation product; and the culture time is 15-24h.

[0012] Furthermore, the preparation process of Bacillus subtilis culture and Lactobacillus plantarum culture in step (1) is as follows: Bacillus subtilis and Lactobacillus plantarum are activated and cultured respectively to obtain Bacillus subtilis culture and Lactobacillus plantarum culture.

[0013] Further, the specific steps of the activation culture are as follows: Bacillus subtilis is inoculated into LB medium and cultured at 37°C for 24-28 hours to obtain Bacillus subtilis bacterial solution; Lactobacillus plantarum is inoculated into MRS medium and cultured at 37°C for 24-28 hours to obtain Lactobacillus plantarum bacterial solution.

[0014] Furthermore, the effective viable count of the Bacillus subtilis bacterial solution is 1-3 × 10⁻⁶. 8 CFU / mL; the effective viable count of the *Lactobacillus plantarum* bacterial suspension is 1-5 × 10⁻⁶. 8 CFU / mL.

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

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

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

[0018] Weigh out the compound bacterial powder, humic acid, and bentonite according to the stated weight proportions, and stir evenly at room temperature to obtain the final product.

[0019] Compared with the prior art, the main advantages of the present invention are:

[0020] This invention improves the stability of compound bacterial powder by adding grain insect powder and corn gluten to the fermentation product, thus maintaining the good activity of microorganisms in the compound bacterial powder and promoting bacterial proliferation. Combined with humic acid and bentonite, it can significantly improve water quality, effectively reduce the concentration of ammonia nitrogen, total nitrogen, total phosphorus, nitrite and other substances in the water, and improve the ecological environment of aquaculture waters. Detailed Implementation

[0021] The technical solution of the present invention will be further described below with reference to specific embodiments. However, those skilled in the art should understand that the following embodiments are only for illustrating the present invention and should not be regarded as limiting the present invention. Specific conditions not specified in the embodiments are performed according to conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the reagents or instruments used are all conventional products obtained through commercial channels.

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

[0023] The grain weevils were dried at 80°C for 4 hours. The dried grain weevils were then pulverized in a grinder for 30 minutes to obtain the grain weevils powder used in this embodiment.

[0024] The Lactobacillus plantarum accession number of this invention is CGMCC1.557; the Bacillus subtilis accession number is CGMCC1.921.

[0025] Example 1

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

[0027] The preparation method of the compound bacterial powder specifically includes 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 Bacillus subtilis bacterial suspension; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 26 h to obtain Lactobacillus plantarum bacterial suspension; the effective viable count of the Bacillus subtilis bacterial suspension was 2 × 10⁻⁶. 8 CFU / mL; the effective viable count of the *Lactobacillus plantarum* bacterial suspension was 3 × 10⁻⁶. 8 CFU / mL.

[0029] (2) Add 15 parts glucose, 25 parts soybean residue, 3 parts potassium dihydrogen phosphate, 3 parts calcium carbonate, 7 parts sodium chloride and 2 parts magnesium sulfate to 50 parts distilled water to prepare a fermentation medium.

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

[0031] (4) Add grain insect powder and zein to the fermentation product of step (3). 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, centrifuge, remove the supernatant, freeze dry, and obtain compound bacterial powder.

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

[0033] Weigh out the compound bacterial powder, humic acid, and bentonite according to the stated weight proportions, and stir evenly at room temperature to obtain the final product.

[0034] Example 2

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

[0036] The preparation method of the compound bacterial powder specifically includes 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 Bacillus subtilis bacterial suspension; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 24 h to obtain Lactobacillus plantarum bacterial suspension; the effective viable count of the Bacillus subtilis bacterial suspension was 1×10⁻⁶. 8 CFU / mL; the effective viable count of the *Lactobacillus plantarum* bacterial suspension is 1×10⁻⁶. 8 CFU / mL.

[0038] (2) Add 10 parts glucose, 20 parts soybean residue, 1 part potassium dihydrogen phosphate, 1 part calcium carbonate, 5 parts sodium chloride and 1 part magnesium sulfate to 40 parts distilled water to prepare fermentation culture medium.

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

[0040] (4) Add grain insect powder and zein to the fermentation product of step (3). The amount of grain insect powder added is twice the total weight of the fermentation product; the amount of zein added is once the total weight of the fermentation product. After culturing for 15 hours, centrifuge, remove the supernatant, freeze dry, and obtain compound bacterial powder.

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

[0042] Weigh out the compound bacterial powder, humic acid, and bentonite according to the stated weight proportions, and stir evenly at room temperature to obtain the final product.

[0043] Example 3

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

[0045] The preparation method of the compound bacterial powder specifically includes 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 Bacillus subtilis bacterial suspension; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 28 h to obtain Lactobacillus plantarum bacterial suspension; the effective viable count of the Bacillus subtilis bacterial suspension was 3 × 10⁻⁶. 8 CFU / mL; the effective viable count of the *Lactobacillus plantarum* bacterial suspension was 5 × 10⁻⁶. 8 CFU / mL.

[0047] (2) By weight, add 20 parts glucose, 30 parts soybean residue, 5 parts potassium dihydrogen phosphate, 5 parts calcium carbonate, 10 parts sodium chloride and 4 parts magnesium sulfate to 60 parts distilled water to prepare a fermentation culture medium.

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

[0049] (4) Add grain insect powder and zein to the fermentation product of step (3). 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, centrifuge, remove the supernatant, freeze dry, and obtain compound bacterial powder.

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

[0051] Weigh out the compound bacterial powder, humic acid, and bentonite according to the stated weight proportions, and stir evenly at room temperature to obtain the final product.

[0052] Comparative Example 1

[0053] The difference between Comparative Example 1 and Example 1 is that the compound bacterial powder prepared in step (4) omits the grain insect powder.

[0054] Comparative Example 2

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

[0056] Comparative Example 3

[0057] The difference between Comparative Example 3 and Example 1 is that the five-grain insect powder is omitted from the compound bacterial powder prepared in step (4), and the amount of corn gliadin is adjusted to be the sum of the two. Everything else is the same as in Example 1.

[0058] Comparative Example 4

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

[0060] Experimental Example 1

[0061] The following are experiments on the effects of the compositions obtained in Examples 1-3 and Comparative Examples 1-4 on improving aquaculture water quality:

[0062] 14L of aquaculture water was collected and divided into four groups: 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 aquaculture water quality improvement composition prepared in Examples 1-3 and Comparative Examples 1-4 was added to each group, once every three days, for three consecutive weeks. After adding the aquaculture water quality improvement composition, the COD and NH4 levels of the water were measured. + -N, total nitrogen, total phosphorus, NO2 - The content of -N. The content of each component in the water before the experiment is shown in Table 1, and the content of each component in the water after adding the composition to improve the aquaculture water quality is shown in Table 2.

[0063] Table 1. Content of various components in the water before the experiment

[0064]

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

[0066]

[0067] The results are shown in Tables 1 and 2, which show the content of each component in the water before the experiment and the content of each component in the water after adding the aquaculture water quality improvement composition, respectively. As can be seen from the tables, after three weeks of continuous feeding, compared with Comparative Examples 1-4, the aquaculture water quality improvement compositions prepared in Examples 1-3 of this invention can significantly reduce the COD and NH4+ content in the water. + -N content, total nitrogen content, total phosphorus content, NO2 - -N content. This indicates that the bentonite, humic acid, and compound bacterial powder in the composition for improving aquaculture water quality prepared in this invention, when used in combination, can effectively reduce the concentrations of ammonia nitrogen, total nitrogen, total phosphorus, nitrite, etc. in the water, thereby improving the ecological environment of aquaculture waters.

[0068] This is because bentonite has a strong adsorption capacity, directly adsorbing toxic substances such as ammonia nitrogen and nitrite. Humic acid can improve aquaculture water quality, promote the healthy growth of aquatic products, reduce the use of chemicals, and enhance the self-purification capacity of the water body. By adding grain insect powder and zein to the fermentation products, the stability of the compound bacterial powder can be effectively improved, ensuring the microorganisms in the compound bacterial powder maintain good activity, promoting bacterial proliferation, and reducing the concentration of ammonia nitrogen, total nitrogen, total phosphorus, nitrite, etc. in the water.

[0069] Further analysis revealed that, compared to Example 1, the compound bacterial powder prepared in Comparative Example 1 omitted the grain insect powder; the compound bacterial powder prepared in Comparative Example 2 omitted zein; the compound bacterial powder prepared in Comparative Example 3 omitted the grain insect powder, and the amount of zein was adjusted to the sum of the two; the compound bacterial powder prepared in Comparative Example 4 omitted zein, and the amount of grain insect powder was adjusted to the sum of the two. The effectiveness of all four groups in improving the ecological environment of aquaculture waters was significantly reduced. This indicates that the grain insect powder and zein in the compound bacterial powder have a synergistic effect, and their combined use is necessary to effectively improve the stability of the compound bacterial powder, maintain the good activity of the microorganisms in the compound bacterial powder, promote bacterial proliferation, effectively reduce the concentrations of ammonia nitrogen, total nitrogen, total phosphorus, and nitrite in the water, and improve the ecological environment of aquaculture waters.

[0070] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. The basic principles and main features of the present invention have been described above with specific implementation schemes. Based on the present invention, some modifications or substitutions can be made, but these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate 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 compound microbial powder, 8-15 parts of humic acid, and 3-9 parts of bentonite; the preparation process of the compound microbial powder is as follows: (1) Inoculate Bacillus subtilis culture and Lactobacillus plantarum culture into fermentation medium and carry out fermentation culture to obtain fermentation product; the Bacillus subtilis culture, Lactobacillus plantarum culture and fermentation medium are inoculated at a volume ratio of 5:(5-9):100; the fermentation culture time is 20-30h; (2) Add grain insect powder and zein to the fermentation product, culture, centrifuge, remove the supernatant, freeze dry to obtain compound bacterial powder; 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; the culture time is 15-24h.

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

3. The composition for improving aquaculture water quality according to claim 2, characterized in that, The specific steps of the activation culture are as follows: Bacillus subtilis is inoculated into LB medium and cultured at 37°C for 24-28 hours to obtain Bacillus subtilis bacterial solution; Lactobacillus plantarum is inoculated into MRS medium and cultured at 37°C for 24-28 hours to obtain Lactobacillus plantarum bacterial solution.

4. The composition for improving aquaculture water quality according to claim 2, characterized in that, The effective viable count of the Bacillus subtilis bacterial suspension was 1×10⁻⁶. 8 CFU / mL up to 3×10 8 CFU / mL; the effective viable count of the *Lactobacillus plantarum* bacterial suspension is 1×10⁻⁶. 8 CFU / mL up to 5×10 8 CFU / mL.

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

6. The method for preparing the composition for improving aquaculture water quality according to any one of claims 1-5, characterized in that, The specific preparation method is as follows: Weigh out the compound bacterial powder, humic acid, and bentonite according to the stated weight proportions, and stir evenly at room temperature to obtain the final product.

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