Preparation with functions of promoting growth of aquatic animals and improving water quality and application thereof

Abstract

The invention discloses a preparation with functions of promoting growth of aquatic animals and improving water quality and application of the preparation. The preparation comprises compound bacterial powder and a traditional Chinese medicine extract in a mass ratio of (2-3): (1-2), the compound bacterial powder is bacillus velezensis J-1 bacterial powder and bifidobacterium G-1 bacterial powder in a mass ratio of (2-4): (3-6), the effective viable count of the bacillus velezensis J-1 bacterial powder is 1.5 * 10 < 9 >-2.5 * 10 < 9 > CFU / g, the effective viable count of the bifidobacterium G-1 bacterial powder is 1.0 * 10 < 10 >-1.5 * 10 < 10 > CFU / g, the bacillus velezensis J-1 and the bifidobacterium G-1 are both preserved in the China Center for Type Culture Collection, and the preservation number of the bacillus velezensis J-1 bacterial powder and the bifidobacterium G-1 bacterial powder is CGMCC (China Center for Type Culture Collection). The preservation number of the bacillus velezensis J-1 is CCTCC (China Center for Type Culture Collection) No: M2021287, and the preservation number of the bifidobacterium G-1 is CCTCC No: M2024350; the traditional Chinese medicine extract is composite extract powder of rhizoma acori graminei and perilla leaves. According to the preparation, the synergistic interaction effect of the two bacteria is utilized, the effects of promoting growth of the aquatic animals and improving water quality can be remarkably improved, and meanwhile the effects of promoting growth of the aquatic animals and improving water quality can be further achieved by adding the compound traditional Chinese medicine extract of the rhizoma acori graminei and the perilla leaves.

Description

Technical Field

[0001] The present invention relates to the technical field of promoting the growth of aquatic animals and improving water quality, and particularly relates to a preparation with the functions of promoting the growth of aquatic animals and improving water quality and its application. Background Art

[0002] In the aquaculture water body of aquatic animals, the content of pathogenic bacteria and viruses is extremely high, which will lead to the imbalance of the flora of aquatic animals and the water body, making the disease resistance of the cultured aquatic animals weaker and the number of drug-resistant strains increasing, seriously affecting the aquaculture level of aquatic animals, and causing food and biosafety problems due to the residue of harmful substances. At present, a large number of microorganisms have been recorded in the relevant literature to have antibacterial effects and are applied to aquaculture and planting.

[0003] Bacillus velezensis J-1 with the preservation number of CCTCC No: M2021287 can produce secondary metabolites that inhibit molds and degrade mycotoxins after fermentation, and has good antibacterial activity and anti-stress ability. As a feed additive, it has many effects such as improving the microbial cell structure of the intestinal tract of livestock and poultry, regulating the body's lipid metabolism, enhancing immunity, and promoting feed digestion and absorption.

[0004] Bifidobacterium G-1 with the preservation number of CCTCC No: M2024350 can reduce the absorption of harmful bacteria, improve digestion, and adjust the intestinal flora; it can form a biological barrier to prevent the colonization and invasion of pathogenic bacteria; regulate the body's immunity; protect the liver and so on. As a feed additive, it can prevent and treat various diseases and protect the health of the body.

[0005] Although the above two kinds of bacteria have good antibacterial effects, so far, there is no report on the application of the mixture of the two kinds of bacteria in aquaculture to significantly improve the effects of promoting the growth of aquatic animals and improving water quality. Summary of the Invention

[0006] The present invention provides a preparation with the functions of promoting the growth of aquatic animals and improving water quality and its application to solve the above problems.

[0007] To achieve the above purpose, the technical solution of the present invention is:

[0008] A preparation with the functions of promoting the growth of aquatic animals and improving water quality, comprising a compound bacterial powder and a traditional Chinese medicine extract with a mass ratio of 2-3:1-2;

[0009] The composite bacterial powder is Bacillus velezensis J-1 bacterial powder and Bifidobacterium G-1 bacterial powder with a mass ratio of 2-4:3-6. The effective viable count of the Bacillus velezensis J-1 bacterial powder is 1.5×10 9 -2.5×10 9 CFU / g, and the effective viable count of the Bifidobacterium G-1 bacterial powder is 1.0×10 10 -1.5×10 10 CFU / g. Both Bacillus velezensis J-1 and Bifidobacterium G-1 have been deposited in the China Center for Type Culture Collection. The deposit number of Bacillus velezensis J-1 is CCTCC No: M2021287, the deposit date of Bacillus velezensis J-1 strain is March 29, 2021, the depository is the China Center for Type Culture Collection, and the deposit address is No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province; the deposit number of Bifidobacterium G-1 is CCTCC No: M2024350, the deposit date of Bifidobacterium G-1 strain is March 7, 2024, the depository is the China Center for Type Culture Collection, and the deposit address is No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province;

[0010] The traditional Chinese medicine extract is a composite extract powder of Acorus tatarinowii and Perilla frutescens leaves.

[0011] Furthermore, the Bacillus velezensis J-1 bacterial powder is prepared according to the following steps:

[0012] After activating and culturing Bacillus velezensis J-1, inoculate it into the first-stage seed medium, then inoculate the first-stage seed into the second-stage seed medium, the second-stage seed into the third-stage seed medium, and the third-stage seed into the fermentation medium. After fermentation, obtain the Bacillus velezensis J-1 fermentation broth. Centrifuge and concentrate the Bacillus velezensis J-1 fermentation broth to obtain the Bacillus velezensis J-1 concentrate; use soluble additive powder to coat the Bacillus velezensis J-1 concentrate to obtain the Bacillus velezensis J-1 bacterial powder.

[0013] Furthermore, the specific preparation steps of the Bacillus velezensis J-1 bacterial powder are as follows:

[0014] Inoculate Bacillus velezensis J-1 onto the NA solid medium (33 g of the medium is dissolved in 1 L of purified water, add 5 g of agar powder, and adjust the pH to 7.0±0.2) for activation and culture in a 37°C incubator for 18-24 h;

[0015] Inoculate the Bacillus velezensis J-1 obtained by solid culture into the first-stage seed medium, and ferment and culture at 35-37°C and 150-160 r / min for 12-18 h to obtain the Bacillus velezensis J-1 first-stage seed liquid;

[0016] Inoculate the first-stage seed liquid of Bacillus velezensis J-1 into the second-stage seed liquid medium, and perform fermentation culture at 35-37 °C and 150-160 r / min for 12-18 h to obtain the second-stage seed liquid of Bacillus velezensis J-1;

[0017] Inoculate the second-stage seed liquid of Bacillus velezensis J-1 into the third-stage seed liquid medium, and perform fermentation culture at 35-37 °C and 150-160 r / min for 12-18 h to obtain the third-stage seed liquid of Bacillus velezensis J-1;

[0018] Each liter of the first-stage seed liquid medium, second-stage seed liquid medium and third-stage seed liquid medium contains 10 g of soy peptone, 3 g of beef extract, 0.5 g of yeast powder, 4 g of glucose, 1.5 g of dipotassium hydrogen phosphate, and is made up to volume with distilled water, and the pH is 5.4-6.5;

[0019] Inoculate the third-stage seed liquid of Bacillus velezensis J-1 into a fermenter, and perform fermentation culture at 37 °C and 150-160 r / min for 18-24 h to obtain the fermentation broth of Bacillus velezensis J-1;

[0020] Use soluble carrier powder to coat the concentrated liquid of Bacillus velezensis J-1 to obtain the bacterial powder of Bacillus velezensis J-1.

[0021] Furthermore, the bacterial powder of Bifidobacterium G-1 is prepared according to the following steps:

[0022] After activating and culturing Bifidobacterium G-1, inoculate it into the first-stage seed medium, then inoculate the first-stage seed into the second-stage seed medium, the second-stage seed into the third-stage seed medium, and the third-stage seed into the fermentation medium. After fermentation, obtain the fermentation broth of Bifidobacterium G-1. After centrifuging and concentrating the fermentation broth of Bifidobacterium G-1, obtain the concentrated liquid of Bifidobacterium G-1; Mix and coat the concentrated liquid of Bifidobacterium G-1 with soluble carrier powder to prepare the bacterial powder of Bifidobacterium G-1.

[0023] Furthermore, the specific preparation steps of the bacterial powder of Bifidobacterium G-1 are as follows:

[0024] Inoculate Bifidobacterium G-1 onto MRS solid medium for activation, and perform anaerobic culture in a 37 °C incubator for 18-24 h;

[0025] Inoculate the Bifidobacterium G-1 obtained by solid culture into the seed medium, and perform anaerobic static fermentation culture at 35-37 °C for 12-18 h to obtain the first-stage seed liquid of Bifidobacterium G-1;

[0026] Inoculate the first-stage seed liquid of Bifidobacterium G-1 into the second-stage seed liquid culture medium, and carry out anaerobic static fermentation culture at 35-37 °C for 12-18 h to obtain the second-stage seed liquid of Bifidobacterium G-1;

[0027] Inoculate the second-stage seed liquid of Bifidobacterium G-1 into the third-stage seed liquid culture medium, and carry out static culture at 35-37 °C for 12-18 h to obtain the third-stage seed liquid of Bifidobacterium G-1;

[0028] Inoculate the third-stage seed liquid of Bifidobacterium G-1 into a fermenter, and carry out anaerobic static fermentation culture at 35-37 °C for 18-24 h to obtain the fermentation broth of Bifidobacterium G-1;

[0029] Centrifuge the fermentation broth at 6000-8000 r / min for 10-20 min and concentrate it by 4 times to obtain the concentrated liquid of Bifidobacterium G-1;

[0030] Mix and coat the concentrated liquid of Bifidobacterium G-1 with soluble beneficial powder to prepare the bacterial powder of Bifidobacterium G-1.

[0031] Furthermore, the compound extract of Acorus tatarinowii and Perilla frutescens is prepared according to the following method:

[0032] S1: Weigh the dried and pulverized Acorus tatarinowii and Perilla frutescens according to a mass ratio of 1.5:1, add purified water to soak for 24 h, carry out ultrasonic treatment, and filter to obtain the extract and the residue;

[0033] S2: Add the residue to the fermentation medium with an inoculum amount of 5% of Bifidobacterium G-1 at a ratio of 5% for anaerobic fermentation, collect the first fermentation extract and the first residue; the viable count of the inoculated Bifidobacterium G-1 is 1.0×10 10 ~1.5×10 10 CFU / mL;

[0034] S3: Add the first residue to the fermentation medium with an inoculum amount of 5% of Bacillus beijerinckii J-1 at a ratio of 5% for anaerobic fermentation, collect the second fermentation extract, and the viable count of Bacillus beijerinckii J-1 is 1.0×10 10 ~1.5×10 10 CFU / mL;

[0035] S4: Mix the extract obtained in S1, the first fermentation extract obtained in S2, and the second fermentation extract obtained in S3 and concentrate them to a crude drug concentration of 1-2 g / mL, and then mix the concentrated liquid with soluble beneficial powder according to a volume-to-mass ratio of 2:1 (mL:g) to prepare the compound extract powder of Acorus tatarinowii and Perilla frutescens.

[0036] Furthermore, it also includes soluble synergistic powder, and the mass ratio of the compound bacterial powder, the compound extract powder of Acorus tatarinowii and Perilla frutescens, and the soluble synergistic powder is 2-3:1-2:5-7.

[0037] Furthermore, the composition of the fermentation medium is as follows: per thousand milliliters contains 15 g of soy peptone, 3 g of yeast powder, 5 g of beef extract, 2.5 g of glucose, 1 mL of Tween 80, 1.5 g of dipotassium hydrogen phosphate, 0.15 g of magnesium sulfate, 0.05 g of manganese sulfate, 5 g of anhydrous sodium acetate, 2.5 g of trisodium citrate, fixed volume with distilled water, and the pH is 5.4-6.5.

[0038] Furthermore, in S2, the conditions for anaerobic fermentation are: 100 r / min, ferment for 18-24 h.

[0039] Furthermore, in S3, the conditions for anaerobic fermentation are: ferment at 160 r / min for 36-48 h.

[0040] On the other hand, the present invention provides the application of the preparation with the functions of promoting the growth of aquatic animals and improving water quality in promoting the growth of aquatic animals.

[0041] On yet another aspect, the present invention provides the application of the preparation with the functions of promoting the growth of aquatic animals and improving water quality in improving water quality.

[0042] The beneficial effects of the present invention are:

[0043] A preparation with the functions of promoting the growth of aquatic animals and improving water quality according to the present invention is a quality mixture of the bacterial powders of Bacillus velezensis J-1 and Bifidobacterium G-1. Utilizing the synergistic effect of the two bacteria, it can inhibit Escherichia coli, aquatic pathogenic bacteria, molds, etc. in the water body, avoid having an adverse impact on the water body, and at the same time reduce the disease infection rate of aquatic animals. Applied in the field of aquaculture, it can significantly improve the efficacy of promoting the growth of aquatic animals and improving water quality. At the same time, adding the compound extract of Acorus tatarinowii and Perilla frutescens can further improve the efficacy of promoting the growth of aquatic animals and improving water quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0045] Figure 1 It is a photo of the effect of Bifidobacterium G-1 of the present invention on aquatic pathogenic bacteria;

[0046] Figure 2 Photograph of the inhibitory effect of the Bifidobacterium G-1 fermentation broth of the present invention on Escherichia coli;

[0047] Figure 3 Photograph of the inhibitory effect of the Bifidobacterium G-1 fermentation broth of the present invention on molds, where Figure 3 a is the experimental result graph of the antibacterial effect of Bifidobacterium on Fusarium moniliforme, Figure 3 b is the experimental result graph of the antibacterial effect of Bifidobacterium on Fusarium, Figure 3 c is the experimental result graph of the antibacterial effect of Bifidobacterium on Fusarium nivale;

[0048] Figure 4 Photograph of the 30-minute flocculation rate result of the Bacillus velezensis J-1 of the present invention in treating kaolin suspension;

[0049] Figure 5 Photograph of the 24-hour flocculation rate result of the Bacillus velezensis J-1 of the present invention in treating kaolin suspension;

[0050] Figure 6 Photograph of the 30-minute flocculation rate result of the Bifidobacterium G-1 of the present invention in treating kaolin suspension;

[0051] Figure 7 Photograph of the 24-hour flocculation rate result of the Bifidobacterium G-1 of the present invention in treating kaolin suspension;

[0052] Figure 8 Flocculation effect result of treating kaolin suspension after mixing the Bacillus velezensis and Bifidobacterium stock solutions of the present invention, where Figure 8 a is the photograph of the kaolin suspension 30 minutes after adding the sample, Figure 8 b is the photograph of the kaolin suspension 24 hours after adding the sample;

[0053] Figure 9 Flocculation effect result of treating kaolin suspension with the preparation of Example 1 of the present invention, where Figure 9 a is the photograph of the kaolin suspension 30 minutes after adding the sample, Figure 9 b is the photograph of the kaolin suspension 24 hours after adding the sample. Detailed implementation mode

[0054] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0055] Preparation Example:

[0056] Preparation Example 1:

[0057] To prepare the Bacillus velezensis J-1 bacterial powder, the following steps are included:

[0058] Step 1: Inoculate Bacillus velezensis J-1 on an NA solid medium (33 g of the medium is dissolved in 1 L of purified water, 5 g of agar powder is added, and the pH is adjusted to 7.0 ± 0.2) for activation, and culture it in an incubator at 37 °C for 18 - 24 h;

[0059] Step 2: Inoculate the Bacillus velezensis J-1 obtained by solid culture into the seed medium, and ferment and culture it at 35 - 37 °C and 150 - 160 r / min for 12 - 18 h to obtain the first-stage seed liquid of Bacillus velezensis J-1;

[0060] Step 3: Inoculate the first-stage seed liquid of Bacillus velezensis J-1 into the second-stage seed liquid medium, and ferment and culture it at 35 - 37 °C and 150 - 160 r / min for 12 - 18 h to obtain the second-stage seed liquid of Bacillus velezensis J-1;

[0061] Step 4: Inoculate the second-stage seed liquid of Bacillus velezensis J-1 into the third-stage seed liquid medium, and ferment and culture it at 35 - 37 °C and 150 - 160 r / min for 12 - 18 h to obtain the third-stage seed liquid of Bacillus velezensis J-1;

[0062] Step 5: Inoculate the third-stage seed liquid of Bacillus velezensis J-1 into a fermenter, and ferment and culture it at 37 °C and 150 - 160 r / min for 18 - 24 h to obtain the fermentation broth of Bacillus velezensis J-1;

[0063] The compositions of the first-stage seed liquid medium, the second-stage seed liquid medium, and the third-stage seed liquid medium are the same, and each contains: 10 g of soy peptone, 3 g of beef extract, 0.5 g of yeast powder, 4 g of glucose, 1.5 g of dipotassium hydrogen phosphate per liter, made up to volume with distilled water, and the pH is 5.4 - 6.5;

[0064] The composition of the fermentation medium (‰) is: 25 g of soluble starch, 30 g of soy peptone, 1 g of yeast powder, 1 g of dipotassium hydrogen phosphate, 0.08 g of magnesium sulfate, 0.05 g of manganese sulfate, made up to 1000 ml with distilled water, and the pH is 5.4 - 6.5.

[0065] The fermentation broth of Bacillus velezensis J-1 was centrifuged at 6000 - 8000 r / min for 10 - 20 min and concentrated 4-fold to obtain a concentrated solution. After coating it with soluble carrier powder, Bacillus velezensis J-1 powder was prepared. Among them, the effective viable count of Bacillus velezensis J-1 powder was 2.5×10 9 CFU / g.

[0066] Preparation Example 2:

[0067] To prepare Bifidobacterium G-1 powder, the following steps are included:

[0068] Step 1: Inoculate Bifidobacterium G-1 onto MRS solid medium for activation and anaerobically culture it in an incubator at 37°C for 18 - 24 h;

[0069] Step 2: Inoculate the Bifidobacterium G-1 obtained from solid culture into the seed medium and anaerobically statically ferment and culture it at 35 - 37°C for 12 - 18 h to obtain the first-stage seed liquid of Bifidobacterium G-1;

[0070] Step 3: Inoculate the first-stage seed liquid of Bifidobacterium G-1 into the second-stage seed liquid medium and anaerobically statically ferment and culture it at 35 - 37°C for 12 - 18 h to obtain the second-stage seed liquid of Bifidobacterium G-1;

[0071] Step 4: Inoculate the second-stage seed liquid of Bifidobacterium G-1 into the third-stage seed liquid medium and statically culture it at 35 - 37°C for 12 - 18 h to obtain the third-stage seed liquid of Bifidobacterium G-1;

[0072] Step 5: Inoculate the third-stage seed liquid of Bifidobacterium G-1 into a fermenter and anaerobically statically ferment and culture it at 35 - 37°C for 18 - 24 h to obtain the fermentation broth of Bifidobacterium G-1;

[0073] The compositions of the first-stage seed liquid medium, the second-stage seed liquid medium, and the third-stage seed liquid medium are the same, and each contains: 10 g of soy peptone, 3 g of beef extract, 0.5 g of yeast powder, 4 g of glucose, 1.5 g of dipotassium hydrogen phosphate per liter, made up to volume with distilled water, and the pH is 5.4 - 6.5;

[0074] The composition of the fermentation medium: 15 g of soy peptone, 3 g of yeast powder, 5 g of beef extract, 2.5 g of glucose, 1 mL of Tween 80, 1.5 g of dipotassium hydrogen phosphate, 0.15 g of magnesium sulfate, 0.05 g of manganese sulfate, 5 g of anhydrous sodium acetate, 2.5 g of trisodium citrate per liter, made up to volume with distilled water, and the pH is 7.0 - 7.2;

[0075] Step 6: The Bifidobacterium G-1 fermentation broth was centrifuged at 6000-8000 r / min for 10-20 min and concentrated 4 times to obtain a concentrate. The concentrate was coated with soluble Jiayi powder according to conventional technology to prepare Bifidobacterium G-1 powder, wherein the effective viable count of Bifidobacterium G-1 powder was 1.5×10 10 CFU / g.

[0076] Preparation Example 3:

[0077] The preparation of the composite bacterial powder of Bacillus Velez and Bifidobacterium comprises the following steps:

[0078] Bacillus Velez J-1 powder and Bifidobacterium G-1 powder are taken in a mass ratio of 2-4:3-6, and are fully mixed to obtain composite powder of Bacillus Velez and Bifidobacterium.

[0079] Preparation Example 4:

[0080] Preparation of a composite extract powder of Acorus calamus and Perilla frutescens leaves comprises the following steps:

[0081] Weigh dried and crushed Acorus calamus and Perilla frutescens leaves at a mass ratio of 1.5:1, add 25 times the mass concentration of purified water, soak for 24 hours, ultrasonically treat for 30 minutes, extract at 65-75°C for 1.5 hours, and obtain an extract and a Chinese medicine residue at the same time;

[0082] The above-mentioned Chinese medicine residues were decomposed and extracted again using the Bacillus berrei J-1 and Bifidobacterium G-1 of the present invention: the Chinese medicine residues were added at a ratio of 5% to the Bifidobacterium G-1 inoculation amount of 5% (the number of viable bacteria was 1.5×10 10 CFU / mL) in a fermentation medium (the fermentation medium was sterilized by high pressure at 115°C for 25 minutes before inoculation), and anaerobically fermented at 100 rpm for 18-24 hours, and the fermentation broth (first fermentation broth) and the residue were collected;

[0083] The residue obtained after the above fermentation was further fermented and decomposed, and the above residue was added to the mixture of Bacillus belcheri J-1 with an inoculation amount of 5% (1.5×10 10 -2.5×10 10 CFU / mL) in a fermentation medium (the fermentation medium was autoclaved at 115°C for 25 minutes before inoculation), and fermented at 160 rpm for 36-48 hours, and the fermentation broth (second fermentation broth) was collected;

[0084] Collect the above-mentioned concentrated extract, the first fermentation broth, and the second fermentation broth, concentrate them to a crude drug concentration of 1 g / mL, mix them evenly, and then coat the concentrated solution with soluble carrier powder according to the conventional method. The volume-to-mass ratio of the concentrated solution to the soluble carrier powder is 2:1 (mL:g) to obtain the composite extract powder of Acorus tatarinowii and Perilla frutescens.

[0085] Examples:

[0086] Example 1:

[0087] Take the composite bacterial powder of Bacillus velezensis and Bifidobacterium (the mass ratio of the two is 3:5) prepared in Preparation Example 3, the composite traditional Chinese medicine extract prepared in Preparation Example 4, and soluble carrier powder (carrier), and mix the three according to a mass ratio of 2.5:2:5.5 to form a preparation - 1 with the functions of promoting the growth of aquatic animals and improving water quality.

[0088] Example 2:

[0089] The difference between this example and Example 1 is only that in this example, the mass ratio of the composite bacterial powder of Bacillus velezensis and Bifidobacterium, the composite extract powder of Acorus tatarinowii and Perilla frutescens, and the soluble carrier powder is: 2:1:5.

[0090] Example 3:

[0091] The difference between this example and Example 1 is only that in this example, the mass ratio of the composite bacterial powder of Bacillus velezensis and Bifidobacterium, the composite extract powder of Acorus tatarinowii and Perilla frutescens, and the soluble carrier powder is: 3:2:7.

[0092] Example 4:

[0093] The difference between this example and Example 1 is only that in this example, the mass ratio of Bacillus velezensis powder to Bifidobacterium powder in the composite bacterial powder of Bacillus velezensis and Bifidobacterium is 2:3.

[0094] Example 5:

[0095] The difference between this example and Example 1 is only that in this example, the mass ratio of Bacillus velezensis powder to Bifidobacterium powder in the composite bacterial powder of Bacillus velezensis and Bifidobacterium is 4:7.

[0096] Comparative example:

[0097] Comparative example 1:

[0098] The difference between this comparative example and Example 1 is only that in this comparative example, only the composite bacterial powder of Bacillus velezensis and Bifidobacterium in Preparation Example 3 is added, and the composite extract powder of Acorus tatarinowii and Perilla frutescens is not added.

[0099] Comparative Example 2:

[0100] The difference between this comparative example and Example 1 is only that in this comparative example, instead of using the compound bacterial powder, only the Bacillus velezensis bacterial powder prepared in Preparation Example 1 is added.

[0101] Comparative Example 3:

[0102] The difference between this comparative example and Example 1 is only that in this comparative example, instead of using the compound bacterial powder, only the Bifidobacterium bacterial powder prepared in Preparation Example 2 is added.

[0103] Comparative Example 4:

[0104] The difference between this comparative example and Example 1 is only that in this comparative example, Lactobacillus casei is used to replace Bifidobacterium to form a mixed bacterial powder, that is, the mass ratio of Bacillus velezensis bacterial powder to Lactobacillus casei bacterial powder is 3:5.

[0105] Comparative Example 5:

[0106] The difference between this comparative example and Example 1 is only that in this comparative example, the mass ratio of Bacillus velezensis to Bifidobacterium in the compound bacterial powder of Bacillus velezensis and Bifidobacterium is 2:1.

[0107] Comparative Example 6:

[0108] The difference between this comparative example and Example 1 is only that in this comparative example, Bacillus velezensis and Bifidobacterium are not used to sequentially treat the residue during the process of compounding the traditional Chinese medicine extract. The specific steps are as follows:

[0109] Acorus tatarinowii and Perilla frutescens are weighed according to a mass ratio of 1.5:1, dried and pulverized, 25 times the mass concentration of purified water is added, soaked for 24 h, ultrasonically treated for 30 min, and then extracted at 65 - 75 °C for 1.5 h. After obtaining the compound traditional Chinese medicine extract, it is concentrated to a raw drug concentration of 1 g / mL.

[0110] Comparative Example 7:

[0111] The difference between this comparative example and Example 1 is only that in this comparative example, the mass ratio of the compound bacterial powder of Bacillus velezensis and Bifidobacterium, the compound traditional Chinese medicine extract, and the soluble beneficial powder is 0.5:1:1, that is, a low-concentration preparation is formed.

[0112] Comparative Example 8:

[0113] The difference between this comparative example and Example 1 is only that in this comparative example, the mass ratio of the compound bacterial powder of Bacillus velezensis and Bifidobacterium, the compound traditional Chinese medicine extract, and the soluble beneficial powder is 3:3:4, that is, a high-concentration preparation is formed.

[0114] 1. Performance tests were conducted on the preparations obtained in Examples 1-4 and Comparative Examples 1-8:

[0115] Carp with similar body weight, body length and good health were selected. After 2 weeks of domestication, they were randomly divided into 15 groups, with 3 replicates in each group and 30 carp in each replicate. The 15 groups were the blank control group, Examples 1-4 and Comparative Examples 1-8. Except for the control group, the fish in the remaining groups were fed with feed containing 0.2% of the preparations of each group. The entire test period was 60 days, and each group was raised according to the unified conventional breeding method. Feed was given once every 12 hours, and the feeding amount was 1.5%-2% of the fish body weight. The water temperature was controlled at 26±2°C, and the water was changed every 3 days. All test group sample preparations were prepared in advance according to 15 kg.

[0116] 1. Effects on the growth of cultured carp and the water quality of the culture water body

[0117] All experimental carp were fasted for 12 hours before and after the start of the experiment, and the body weight of each group of carp was weighed. After the experiment ended, the weight gain rate, survival rate and feed coefficient of the carp were calculated. The results are shown in Table 1.

[0118] Table 1 Effects of each example and comparative example on the growth of carp

[0119]

[0120] The results showed that the preparations of the example groups and comparative example groups of the present invention were significantly higher than the control group in terms of the weight gain rate and survival rate of carp, and the feed coefficient was significantly lower than that of the control group. Among them, Example 1 had the best effect, and its weight gain rate increased by 37.9% compared with the control group. The proportion of the weight gain rate of Examples 1-4 higher than that of the comparative example preparation group was between 18.29% and 32.49%;

[0121] By comparing Examples 1-4 with Comparative Example 1, it can be seen that the weight gain rate and survival rate of the preparation group without the addition of traditional Chinese medicine extract are significantly reduced. Therefore, it is indicated that the traditional Chinese medicine extract in this preparation can synergistically act with the compound bacterial powder to promote animal growth. By comparing the results of Examples 1-4 with those of Comparative Example 2 and Comparative Example 3, it can be seen that the groups adding only a single bacterial powder will significantly reduce the weight gain rate and survival rate of carp, and the effect is significantly worse than that of the preparation with the mixed bacterial powder (Example group), proving that Bacillus velezensis and Bifidobacterium have a synergistic effect in promoting animal growth. By comparing the results of Examples 1-4 with those of Comparative Example 4, it can be seen that the effect of the bacterial agent prepared with non-Bifidobacterium in promoting animal growth is much lower than that of the preparation prepared with Bifidobacterium. Therefore, it is further proved that Bacillus velezensis and Bifidobacterium have a synergistic effect in promoting animal growth. From the results of Examples 1-4 and Comparative Example 5, it can be seen that when the proportion of Bifidobacterium powder in the compound bacterial powder is relatively low (the ratio of the two is higher than the scope of this application), its effect in promoting animal growth is also significantly lower than that of the preparations in Examples 1-4. It can be seen that the ratio of Bacillus velezensis and Bifidobacterium needs to be within the scope provided by this application. From the results of Examples 1-4 and Comparative Example 6, it can be seen that without multiple fermentation treatments of the medicinal residues under the condition of inoculating strains, the effect is close to that of Examples 4 and 5, but significantly lower than that of Examples 1-3, proving the importance of the method for preparing the compound traditional Chinese medicine extract in the present invention. From the results of Examples 1-4 and Comparative Examples 7 and 8, it can be seen that the effects of the low-concentration ratio preparation and the high-concentration ratio preparation of the mixed bacterial powder of Bacillus velezensis and Bifidobacterium, the compound traditional Chinese medicine extract and the soluble beneficial powder are equivalent to those of Examples 3 and 2, but significantly worse than that of Example 1.

[0122] 2. Influence on the water quality of the aquaculture water body

[0123] Water sample collection and pretreatment: After a full water change was carried out during the experiment period, the aquaculture was carried out according to the conventional feeding method, and normal feeding was carried out during this period. The water samples were collected for the first time before grouping and feeding, and the second and third water samples were collected at 24 h and 48 h after the preparation was sprinkled respectively. The water was taken 15 cm away from the water surface at each sample point. The collected water samples were immediately placed in an ice box for preservation and detection, and the results are shown in Table 2.

[0124] Table 2 Influence of the Example group on the aquaculture water body situation

[0125]

[0126] From the results in Table 2, it can be known that the preparations of Examples 1 and 3 of the present invention can significantly maintain the stability of the water quality of the carp aquaculture water body, and can maintain COD and NH4 + -N at a relatively low level, and the pH is maintained at a level more suitable for the growth of carp than that of the control group, better achieving the effect of water body purification and further promoting the growth of carp.

[0127] II. Evaluation of the enzyme-producing ability of Bacillus velezensis used in the examples, and the antibacterial effects of single strains and composite strains of Bacillus velezensis and Bifidobacterium

[0128] 1. For the fermentation broth of Bacillus velezensis J-1 obtained by the fermentation method of the present invention, the enzyme content in the fermentation broth was measured according to the existing method, and the enzyme production effect is shown in Table 3.

[0129] Table 3 Enzyme production of Bacillus velezensis J-1 strain after fermentation

[0130] Detection Index Result β-1,3-1,4-Glucanase (μg / min / mL) 34.58 Alkaline Protease (μg / min / mL) 14.41 Neutral Protease (μg / min / mL) 15.76 Chitinase (μg / h / mL) 89.37 β-Amylase (μg / min / mL) 2158.37 Trypsin (nmol / min / mL) 58.72 Chymotrypsin (nmol / min / mL) 31.09 Lipase (nmol / min / mL) 125.85 Endo-β-1,4-Glucanase (μg / h / mL) 1402.46 Glucose Oxidase (μmol / h·mL) 0.1338 Catalase (μmol / h·mL) 7.21 Manganese Peroxidase (nmol / min / mL) 1.9656 Pepsin (U / mL) 81.07

[0131] Enzymes are divided into endogenous enzymes and exogenous enzymes, which can open plant cell walls and effectively digest feed raw materials for absorption and utilization by the body. Endogenous enzymes are secreted by the body itself and help with the digestion and absorption of chyme; exogenous enzymes can supplement the insufficient secretion and incomplete types of endogenous enzymes, accelerate feed digestion, and promote the absorption of nutrients. As can be seen from Table 3, in the fermentation process of Bacillus velezensis J-1, the optimized process of the present invention can produce neutral protease (Sarre peptidase), alkaline protease (Alkaline protease), pepsin (Pepsin), β-amylase (beta-Amylase), β-1,3-1,4-glucanase, trypsin (Trypsin), chymotrypsin (Chymotrypsin), lipase (LPS), manganese peroxidase (Mnp), glucose oxidase (GOD), catalase (CAT), endo-β-1,4-glucanase (Endo-β-1,4-glucanase), chitinase (Chitinase). Moreover, the types of enzymes produced are numerous, and the contents of β-amylase, endo-β-1,4-glucanase, lipase, and pepsin all occupy the main positions. The rich enzyme system lays a solid foundation for the biological functions of this strain of bacteria.

[0132] 2. Antibacterial effect of Bacillus velezensis J-1

[0133] The Bacillus velezensis J-1 fermentation broth obtained by the fermentation method of Preparation Example 1 of the present invention was centrifuged at 10,000 r / min for 10 min, and then the supernatant was removed and reserved. The Oxford cup method was used for the experiment. Vibrio vulnificus ATCC27562, Vibrio alginolyticus ATCC33787, Escherichia coli CICC10389, Shewanella putrefaciens ATCC BAA-1097, Salmonella typhimurium CICC21513, Staphylococcus aureus CICC10384, Vibrio parahaemolyticus ATCC17802, Aeromonas hydrophila BNCC336453, Vibrio cholerae BNCC 232030 indicator bacteria solutions were of the order of 1×10 8 CFU / mL. 100 μL was taken and evenly spread on a nutrient agar solid plate. Oxford cups were evenly placed. 200 μL of the sample was respectively sucked and put into the Oxford cups according to the serial number, and then placed in an incubator at 37 °C for 24 h. After that, the antibacterial diameter was observed and the results were recorded, as shown in Table 4.

[0134] Table 4 Antibacterial experimental results of Bacillus velezensis

[0135] Detected Bacterial Species Fermentation Bacteriostatic Diameter of the Invention (mm) Escherichia coli 14.2 Salmonella 14.0 Staphylococcus aureus 22.9 Aeromonas hydrophila 13.56 Shewanella alga 13.92 Vibrio alginolyticus 13.8 Vibrio cholerae 12.49 Vibrio vulnificus 12.0

[0136] 3. Effect of Bifidobacterium on aquatic pathogenic bacteria

[0137] After the Bifidobacterium prepared in Preparation Example 2 of the present invention was fermented, three samples of the fermented stock solution, the supernatant, and the 0.45 μm filtrate were obtained respectively. Using Shewanella putrefaciens, Vibrio parahaemolyticus, Vibrio alginolyticus, and Pseudomonas pseudoalteromonad as indicator bacteria, the antibacterial effects were investigated. The results are shown in Table 5 and Figure 1 shown ( Figure 1 a, b, c, and d in

[0138] Table 5 Inhibitory effect of Bifidobacterium fermentation broth treatment group on aquatic pathogenic bacteria mm

[0139] Pathogenic Bacteria Original Solution of Bifidobacterium Fermentation Broth Supernatant after Centrifugation of Fermentation Broth Fermentation Broth after 0.45um Filtration Shewanella <![CDATA[14.99±0.03 Bb > <![CDATA[16.36±0.04 Aa > <![CDATA[14.46±0.04 Aa > Vibrio parahaemolyticus <![CDATA[11.55±0.06 Cc > <![CDATA[10.28±0.02 Cc > <![CDATA[10.28±0.02 Cc > Vibrio alginolyticus <![CDATA[11.05±0.05 Dd > <![CDATA[9.97±0.06 Dd > <![CDATA[9.90±0.02 Dd > Pseudoalteromonas <![CDATA[18.78±0.03 Aa > <![CDATA[15.64±0.03 Bb > <![CDATA[13.50±0.02 Bb >

[0140] As can be seen from Table 5 above, each treatment group of Bifidobacterium has inhibitory effects on Shewanella putrefaciens, Vibrio parahaemolyticus, Vibrio alginolyticus, and Pseudomonas pseudoalteromonad, and the differences are extremely significant. From Figure 1As can be seen from the results (in the figure, labels 1, 2, and 3 are the fermentation broths prepared by the ordinary fermentation process, and labels 4, 5, and 6 are the original bifidobacterium fermentation broths prepared by the multi-stage fermentation of the present invention, the supernatant after centrifugation of the fermentation broth, and the fermentation broth filtered through 0.45 μm), the antibacterial effect of the bifidobacterium fermentation broth prepared by the present invention is significantly higher than that of the ordinary fermentation process.

[0141] 4. Effect of Bifidobacterium Fermentation Broth on Escherichia coli

[0142] After fermentation by the fermentation process of the present invention, three samples of fermentation stock solution, supernatant, and 0.45 μm filtrate were obtained respectively. Using Escherichia coli O1 and O 78 as indicator bacteria, the antibacterial effect was investigated. Samples 1, 2, and 3 are the samples after fermentation by the process of the present invention (1 is the original bifidobacterium fermentation broth; 2 is the supernatant after centrifugation of the fermentation broth; 3 is the fermentation broth after filtration through 0.45 μm).

[0143] Table 6 Inhibitory effect of bifidobacterium fermentation broth treatment group on Escherichia coli mm

[0144]

[0145] From the results in Table 6 and Figure 2 the results, it can be seen that each treatment group of bifidobacterium fermentation broth has an antibacterial effect of medium sensitivity or above on Escherichia coli (O1, O 78 ), and the inhibitory effects of each treatment group of the fermentation broth on the two pathogenic bacteria are extremely significantly different. It fully shows that bifidobacterium can be used for the prevention and control of Escherichia coli in the aquaculture industry.

[0146] 5. Inhibitory effect of bifidobacterium fermentation broth on molds

[0147] After fermentation by the fermentation process of the present invention, three samples of fermentation stock solution, supernatant, and 0.45 μm filtrate were obtained respectively. Using Fusarium oxysporum, Fusarium moniliforme, and Fusarium nivale as indicator bacteria, the antibacterial effect was investigated. Samples 1, 2, and 3 are the samples after fermentation by the process of the present invention (1 is the original bifidobacterium fermentation broth; 2 is the supernatant after centrifugation of the fermentation broth; 3 is the fermentation broth after filtration through 0.45 μm).

[0148] Table 7 Inhibitory effect of bifidobacterium fermentation broth treatment group on molds mm

[0149]

[0150] Figure 3 a is the experimental result diagram of the antibacterial effect of bifidobacterium on Fusarium moniliforme, Figure 3 b is the experimental result diagram of the antibacterial effect of bifidobacterium on Fusarium oxysporum, Figure 3 c is the experimental result diagram of the antibacterial effect of bifidobacterium on Fusarium nivale, where Figure 3 a andFigure 3 The label 25 in b is the original bifidobacterium fermentation broth, 26 is the supernatant after centrifugation of the fermentation broth; 27 is the fermentation broth filtered through 0.45um;

[0151] As can be seen from Table 7 and Figure 3 Among them, each treatment group of bifidobacterium has an inhibitory effect above medium sensitivity on Fusarium nivale, and the inhibitory effects of the original bifidobacterium fermentation broth, the supernatant after centrifugation, and the filtrate on Fusarium nivale are extremely significantly better than those on Fusarium oxysporum and Fusarium moniliforme; the inhibitory effects of the supernatant after centrifugation and the filtrate on Fusarium moniliforme are extremely significant, and the effect is better than that on Fusarium oxysporum.

[0152] 6. Inhibitory effect of the mixture of Bacillus velezensis and Bifidobacterium on aquatic pathogenic bacteria

[0153] Mix the Bacillus velezensis fermentation broth and the Bifidobacterium fermentation broth in a volume ratio of 3:5, that is, mix 3 mL of the original Bacillus velezensis fermentation broth and 5 mL of the original Bifidobacterium fermentation broth to make a sample.

[0154] Table 8 Inhibitory effect of the original mixed fermentation broth on aquatic pathogenic bacteria mm

[0155]

[0156]

[0157] All the above antibacterial results can preliminarily show that the bifidobacterium obtained by the present invention can inhibit the growth of molds and prevent the production of mycotoxins, and can be used in aquaculture and feed additives. At the same time, when Bacillus velezensis and Bifidobacterium are mixed in a ratio of 3:5, they have a combined effect of improving the antibacterial effect.

[0158] III. Evaluation of the flocculation effect of Bacillus velezensis, Bifidobacterium, and the preparation prepared in Example 1

[0159] 1. Preparation of kaolin suspension: (mixed with distilled water)

[0160] In this experiment, the kaolin sedimentation method was used as the selection standard for flocculants, and the suspension was prepared as follows: 4 g of kaolin was added to 1 L of distilled water, and magnetically stirred for 30 min. It was prepared immediately before use, with a pH of 7.2, and the coagulant aid was 1% CaCl2 by mass fraction.

[0161] 2. Determination method of flocculation activity:

[0162] Add 100 mL of the prepared kaolin suspension into a 200 mL beaker, add 2 mL of 1% CaCl₂ solution, adjust the pH to 7.0 - 7.5 with NaOH. Add 2 mL of the strain fermentation broth or cell suspension to each test group, stir at 200 r / min for 3 min, and let it stand for 5 min. Take samples at fixed points (1 cm below the liquid surface) at 30 min and 24 h, measure the absorbance of the supernatant at 550 nm using an ultraviolet spectrophotometer, and calculate the flocculation rate based on the absorbance.

[0163] The calculation formula for the flocculation rate is:

[0164] Flocculation rate = (A - B) / A × 100%

[0165] In the formula, A is the absorbance of the control supernatant at 550 nm; B is the absorbance of the sample supernatant at 550 nm.

[0166] 3. Experimental results:

[0167] The results are as shown in Table 9 - 12 and Figures 4 - 8 as follows:

[0168] Table 9 Flocculation rates of each treatment group of Bacillus velezensis

[0169] Treatment Details Flocculation Rate at 30 min (%) Flocculation Rate at 24 h (%) Bacillus velezensis Culture Medium Control 0 0 Original Solution of Bacillus velezensis (400 million / mL) <![CDATA[62.23±0.19 Bb > <![CDATA[79.94±0.32 Bb > Supernatant of Bacillus velezensis <![CDATA[80.67±0.26 Aa > <![CDATA[83.12±0.32 Aa > Resuspended Bacillus velezensis Cells <![CDATA[54.74±0.75 Cc > <![CDATA[76.11±0.32 Cc >

[0170] As can be seen from Table 9, with the addition of the Bacillus velezensis culture medium as the blank control, the OD values were compared by sampling at 30 min and 24 h after adding the samples. The flocculation rate of the supernatant of the Bacillus velezensis fermentation broth was significantly higher than that of the fermentation broth stock solution and the cell suspension group (p < 0.01), and the fermentation broth stock solution group was significantly higher than the cell suspension group. Figure 4 is a photo of the kaolin suspension 30 min after adding the sample, Figure 5 is a photo of the kaolin suspension 24 h after adding the sample. For the flocculation situation, Bacillus velezensis can achieve an effect of 50% - 80% 30 min after adding the sample, and the flocculation rate slightly increases after 24 h. Therefore, it is proved that the Bacillus velezensis involved in the present invention has a flocculation effect and shows an upward trend with the extension of the action time.

[0171] Table 10 Flocculation rates of each treatment group of Bifidobacterium

[0172] Treatment Details Flocculation Rate at 30 min (%) Flocculation Rate at 24 h (%) Bifidobacterium Culture Medium Control 0 0 Original Solution of Bifidobacterium (3 billion / mL) <![CDATA[7.49±0.37 Cc > <![CDATA[70.02±0.40 Cc > Supernatant of Bifidobacterium <![CDATA[38.44±0.63 Bb > <![CDATA[75.22±1.11 Bb > Resuspended Bifidobacterium Cells <![CDATA[48.45±0.13 Aa > <![CDATA[98.41±0.27 Aa >

[0173] As can be seen from Table 10, with the medium added with Bifidobacterium as the blank control, samples were taken at 30 min and 24 h after adding the samples for OD value comparison. By comparing the results at 30 min and 24 h, it can be seen that for the flocculation rate of Bifidobacterium, the flocculation rate of the resuspended cells was significantly higher than that of the original fermentation broth and its supernatant, the supernatant of the fermentation broth was higher than the original fermentation broth, and with the extension of time, the flocculation rate increased significantly. Figure 6 This is a photo of the kaolin suspension 30 min after adding the sample. Figure 7 This is a photo of the kaolin suspension 24 h after adding the sample. The initial flocculation rate of Bifidobacterium was not very high, and the flocculation rate of the original fermentation broth did not reach 10%, while after acting for 24 h, they all reached over 70%, and even the resuspended cell group reached over 98%.

[0174] Table 11 Flocculation rate of the mixed fermentation broth of two strains

[0175]

[0176] Table 11 shows the flocculation effect results after mixing the original broth of Bacillus velezensis and Bifidobacterium. Figure 8 a is a photo of the kaolin suspension 30 min after adding the sample. Figure 8 b is a photo of the kaolin suspension 24 h after adding the sample. It can be seen that the mixture of the two has a good flocculation effect.

[0177] Table 12 Flocculation rate of the preparation of the present invention

[0178] Treatment Details Flocculation Rate at 30 min (%) Flocculation Rate at 24 h (%) Example 1 53.55 99.50

[0179] Table 12 and Figure 9 This is the flocculation effect and photo of the preparation of Example 1 of the present invention. Figure 9 a is a photo of the kaolin suspension 30 min after adding the sample. Figure 9 b is a photo of the kaolin suspension 24 h after adding the sample. From the data in the table and Figure 9 it can be seen that the preparation in Example 1 of the present invention has a significant flocculation effect. The preparation of the present invention can inhibit pathogenic bacteria in aquaculture water and at the same time has the effect of improving water quality, playing a good role in aquaculture. The main substance that plays a flocculation role in the preparation is the probiotic preparation.

[0180] The Bacillus velezensis and Bifidobacterium involved in the present invention both belong to probiotic microorganisms. In addition to inhibiting Escherichia coli and Salmonella, they can also inhibit the common pathogenic Vibrio in aquaculture and have a flocculation effect, which helps the water quality to quickly recover to cleanliness, degrade harmful substances in the water body, maintain the pH balance of the water body, degrade ammonia nitrogen and COD, and provide a safe and effective water body culture environment for aquatic animals. At the same time, Bacillus velezensis also has the effect of inhibiting molds. Therefore, it not only achieves the effect of preventing and treating diseases, but also synchronously regulates the intestinal flora of aquatic animals, maintains intestinal health and flora balance in both aerobic and anaerobic dimensions, and improves the body's immunity. The two strains act synergistically, and at the same time, the synergistic compound traditional Chinese medicine extract can achieve the effects of antibacterial and improving the animal body's immunity, protecting the body organs from being invaded, etc., and achieve the effects of promoting animal growth and improving water quality.

[0181] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than limiting them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A preparation with the functions of promoting the growth of aquatic animals and improving water quality, characterized in that, It includes a compound bacterial powder with a mass ratio of 2 - 3:1 - 2 and a traditional Chinese medicine extract; The composite bacterial powder is a powder of Bacillus velezensis J-1 and Bifidobacterium G-1 with a mass ratio of 2-4:3-6. The effective viable count of the Bacillus velezensis J-1 bacterial powder is 1.5×10 9 ~2.5×10 9 CFU / g, and the effective viable count of the Bifidobacterium G-1 bacterial powder is 1.0×10 10 ~1.5×10 10 CFU / g. Both Bacillus velezensis J-1 and Bifidobacterium G-1 have been deposited in the China Center for Type Culture Collection. The deposit number of Bacillus velezensis J-1 is CCTCC No: M2021287, and the deposit number of Bifidobacterium G-1 is CCTCC No: M2024350; The traditional Chinese medicine extract is a composite extract powder of Acorus tatarinowii and Perilla frutescens.

2. The preparation with the functions of promoting the growth of aquatic animals and improving water quality according to claim 1, characterized in that, The Bacillus velezensis J-1 bacterial powder is prepared according to the following steps: After activating and culturing Bacillus velezensis J-1, it is inoculated into a first-stage seed medium, then the first-stage seeds are inoculated into a second-stage seed medium, the second-stage seeds are inoculated into a third-stage seed medium, and the third-stage seeds are inoculated into a fermentation medium. After fermentation, a Bacillus velezensis J-1 fermentation broth is obtained. The Bacillus velezensis J-1 fermentation broth is centrifuged and concentrated to obtain a Bacillus velezensis J-1 concentrate; the Bacillus velezensis J-1 concentrate is coated with soluble additive powder to obtain Bacillus velezensis J-1 bacterial powder.

3. The preparation with the functions of promoting the growth of aquatic animals and improving water quality according to claim 2, wherein The Bifidobacterium G-1 bacterial powder is prepared according to the following steps: After activating and culturing Bifidobacterium G-1, it is inoculated into a first-stage seed medium, then the first-stage seeds are inoculated into a second-stage seed medium, the second-stage seeds are inoculated into a third-stage seed medium, and the third-stage seeds are inoculated into a fermentation medium. After fermentation, a Bifidobacterium G-1 fermentation broth is obtained. The Bifidobacterium G-1 fermentation broth is centrifuged and concentrated to obtain a Bifidobacterium G-1 concentrate; the Bifidobacterium G-1 concentrate is mixed and coated with soluble additive powder to prepare Bifidobacterium G-1 bacterial powder.

4. The preparation with the functions of promoting the growth of aquatic animals and improving water quality according to claim 1, characterized in that, The composite extract of Acorus tatarinowii and Perilla frutescens is prepared according to the following method: S1: Weigh dried and pulverized Acorus tatarinowii and Perilla frutescens according to a mass ratio of 1.5:1, add purified water to soak for 24 h, perform ultrasonic treatment, and filter to obtain an extract and residue; S2: Add the residue into the fermentation medium with a Bifidobacterium G-1 inoculum of 5% at a ratio of 5% for anaerobic fermentation, and collect the first fermentation extract and the first residue; the viable count of the inoculated Bifidobacterium G-1 is 1.0×10 10 ~1.5×10 10 CFU / mL; S3: Add the first residue to the fermentation medium with a 5% inoculum of Bacillus beijingensis J-1 at a ratio of 5% for anaerobic fermentation, and collect the second fermentation extract. The viable cell count of Bacillus beijingensis J-1 is 1.0×10 10 ~1.5×10 10 CFU / mL; S4: Mix and concentrate the extract obtained in S1, the first fermentation extract obtained in S2, and the second fermentation extract obtained in S3 to a crude drug concentration of 1 - 2 g / mL, and then coat the concentrated solution with soluble additive powder according to a volume-to-mass ratio of 2:1 (mL:g) to prepare a composite extract powder of Acorus tatarinowii and Perilla frutescens.

5. The preparation with the functions of promoting the growth of aquatic animals and improving water quality according to claim 1, characterized in that, It also includes soluble additive powder, and the mass ratio of the composite bacterial powder, the composite extract powder of Acorus tatarinowii and Perilla frutescens, and the soluble additive powder is 2 - 3:1 - 2:5 - 7.

6. The preparation with the functions of promoting the growth of aquatic animals and improving water quality according to claim 3, characterized in that, The composition of the fermentation medium is: containing 15 g of soy peptone, 3 g of yeast powder, 5 g of beef extract, 2.5 g of glucose, 1 mL of Tween 80, 1.5 g of dipotassium hydrogen phosphate, 0.15 g of magnesium sulfate, 0.05 g of manganese sulfate, 5 g of anhydrous sodium acetate, 2.5 g of trisodium citrate per liter, made up to volume with distilled water, and the pH is 5.4 - 6.

5.

7. The preparation with the functions of promoting the growth of aquatic animals and improving water quality according to claim 4, characterized in that In S2, the conditions for anaerobic fermentation are: 100 r / min, ferment for 18 - 24 h.

8. The preparation with the functions of promoting the growth of aquatic animals and improving water quality according to claim 4, wherein, In S3, the conditions for anaerobic fermentation are: ferment at 160 r / min for 36 - 48 h.

9. Application of the preparation with the functions of promoting the growth of aquatic animals and improving water quality according to claim 1 in promoting the growth of aquatic animals.

10. Application of the preparation with the functions of promoting the growth of aquatic animals and improving water quality according to claim 1 in improving water quality.

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