Bacillus subtilis and its application and products

Abstract

The present invention discloses a strain of Bacillus subtilis and its application and product, and relates to the field of microbial technology. The Bacillus subtilis of the present invention is Bacillus subtilis B‑1158, which has been deposited in the Guangdong Provincial Microbial Culture Collection Center on December 28, 2021, with a deposit number of GDMCC No.62165. The Bacillus subtilis of the present invention has excellent antibacterial activity against Vibrio parahaemolyticus and is resistant to high temperatures, which can solve the problems of low antibacterial rate and poor high temperature resistance of existing Bacillus bacteria against Vibrio parahaemolyticus. At the same time, the Bacillus subtilis of the present invention can also produce amylase and protease, and the antibacterial activity and enzyme production performance of Bacillus subtilis can be used in products that inhibit Vibrio parahaemolyticus. At the same time, it can also be used to prepare water purification products for aquatic products and liver protection products for aquatic products, achieving the effects of antibacterial, water purification and liver protection.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, in particular to a strain of Bacillus subtilis and applications and products thereof. Background Art

[0002] Vibrio parahaemolyticus (VP) is widely present in seawater and seafood. It is a common food poisoning pathogen in coastal areas of my country. It is known that Vibrio parahaemolyticus has 12 O antigens and 59 K antigens. It can be divided into 5 types according to its fermentation of sugars. Various Vibrio species have strong toxicity to humans and animals. Its pathogenic substances mainly include pyrogenic hemolysin (TDH) with a molecular weight of 42,000 and TDH-like hemolytic toxin (TRH) with a molecular weight of 48,000, which have hemolytic activity, enterotoxin and lethal effects.

[0003] Vibrio parahaemolyticus not only harms marine aquaculture organisms such as shrimp, fish, shellfish and other crustaceans, but humans who eat foods containing Vibrio parahaemolyticus will suffer from Vibrio parahaemolyticus food poisoning. The main clinical symptoms of Vibrio parahaemolyticus food poisoning are acute onset, abdominal pain, vomiting, diarrhea and watery stools. The main pathological changes are mild erosion of the jejunum and ileum, gastric mucosal inflammation, and congestion of internal organs (liver, spleen, and lungs). Therefore, inhibiting the activity of Vibrio parahaemolyticus is of great significance to aquatic animals and humans.

[0004] At present, the prevention and treatment of Vibrio parahaemolyticus generally adopts chemical drugs such as bacteriocins and disinfectants for treatment, but drug residues and drug resistance can easily cause environmental pollution and bring harm to humans. With the development of microecological preparations, microecological preparations have become the most potential substitutes for bacteriocins and disinfectants because of their advantages such as green environmental protection, no toxic side effects, and no residual pollution. Probioties, also called live bacteria preparations (Bigone) or probiotics, refer to preparations made by special processes using the principles of microecology and the use of probiotics or probiotic growth-promoting substances that are beneficial and harmless to the host. Probioties can promote the growth and reproduction of normal microorganisms and inhibit the growth and reproduction of pathogenic bacteria. At present, probioties have been used in various fields such as feed, agriculture, medicine and food. 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 are an ideal microecological preparation for inhibiting Vibrio parahaemolyticus. However, the existing disclosed Bacillus bacteria have a low inhibition rate on Vibrio parahaemolyticus and poor high temperature resistance. They are easily inactivated after high temperature treatment, resulting in a significant decrease in antibacterial activity. Summary of the invention

[0005] The purpose of the present invention is to propose a strain of Bacillus subtilis, which is Bacillus subtilis B-1158, which has been deposited in the Guangdong Provincial Microbiological Culture Collection Center on December 28, 2021, with a preservation number of GDMCC No. 62165. The Bacillus subtilis of the present invention has excellent antibacterial activity against Vibrio parahaemolyticus and is resistant to high temperatures, and can solve the problems of low inhibition rate and poor high temperature resistance of existing Bacillus bacteria against Vibrio parahaemolyticus.

[0006] Another object of the present invention is to provide an antibacterial product for aquaculture, which can effectively inhibit Vibrio parahaemolyticus and various aquaculture pathogenic bacteria.

[0007] Another object of the present invention is to provide a water purification product for aquaculture, which can effectively degrade organic matter in aquaculture water, thereby achieving the effect of purifying the water.

[0008] Another object of the present invention is to provide a liver protection product for aquatic products, which can achieve the liver protection effect through the synergistic effect of multiple components including compound bacterial powder, compound traditional Chinese medicine, antimicrobial peptides and yeast nucleotides.

[0009] To achieve this object, the present invention adopts the following technical solutions:

[0010] A strain of Bacillus subtilis, Bacillus subtilis B-1158, was deposited in Guangdong Provincial Microbiological Culture Collection Center on December 28, 2021, with the deposit number GDMCC No.62165.

[0011] Further description is the use of the Bacillus subtilis in the preparation of a product for inhibiting Vibrio parahaemolyticus.

[0012] A bacteriostatic product for aquatic products contains the above-mentioned Bacillus subtilis.

[0013] To further illustrate, the preparation method of the antibacterial product for aquatic products is as follows: Bacillus subtilis and culture medium are prepared into a culture expansion bag to obtain the antibacterial product for aquatic products.

[0014] A water purification product for aquatic products comprises, by weight, 15 to 25 parts of Bacillus subtilis, 15 to 25 parts of lactic acid bacteria, 15 to 25 parts of Bacillus licheniformis and 25 to 55 parts of talc, wherein the Bacillus subtilis is the above-mentioned Bacillus subtilis.

[0015] To further illustrate, the preparation method of the water purification product for aquatic products is as follows: Bacillus subtilis, lactic acid bacteria, Bacillus licheniformis and talcum powder are uniformly mixed to obtain the water purification product for aquatic products.

[0016] A liver protection product for aquatic products, comprising, by weight, 45 to 75 parts of compound bacterial powder, 15 to 25 parts of compound traditional Chinese medicine, 5 to 15 parts of antimicrobial peptides and 5 to 15 parts of yeast nucleotides;

[0017] In terms of mass percentage, the composite bacterial powder comprises 15 to 25 parts of Bacillus subtilis, 15 to 25 parts of lactic acid bacteria and 15 to 25 parts of Bacillus licheniformis, wherein the Bacillus subtilis in the composite bacterial powder is the above-mentioned Bacillus subtilis.

[0018] To further illustrate, the composite Chinese medicine includes salvia miltiorrhiza, astragalus, wolfberry and schisandra chinensis.

[0019] The above technical solution has the following beneficial effects:

[0020] 1. The Bacillus subtilis in the technical solution has antibacterial activity and can inhibit Vibrio parahaemolyticus, Vibrio cholerae, Edwardsiella tarda, Aeromonas sobria and Aeromonas hydrophila. Among them, the Bacillus subtilis in the technical solution has excellent antibacterial activity against Vibrio parahaemolyticus. Moreover, the antibacterial ability of the Bacillus subtilis in the solution has the characteristics of high temperature resistance. After the fermentation liquid of BSW is treated at 80°C, the antibacterial activity can be maintained at about 90%, and after being treated at 100°C, the antibacterial activity can still be maintained at about 70%. Based on the use of BSW in the scope of feed or feed additives, the production process of high-temperature granulation may be involved. The high-temperature resistance of the antibacterial activity of BSW makes it more suitable for high-temperature granulation than other strains, and has a wider range of use. At the same time, because BSW is a bacterium separated from the sludge at the bottom of the shrimp pond, it belongs to the indigenous bacteria in the shrimp pond breeding environment and has a stronger adaptability to the breeding ecology, so that BSW and products containing BSW can adapt to the complex environment of the breeding water body and take effect faster.

[0021] 2. BSW not only has an antibacterial effect, but also can produce amylase and protease. At the same time, Bacillus licheniformis can also produce a variety of digestive enzymes, such as protease, lipase and amylase. At the same time, because lactic acid bacteria are anaerobic bacteria, they can adapt to the low-oxygen environment near the bottom of the pond. This technical solution can effectively degrade the organic matter in the aquaculture water through the complex action of Bacillus subtilis, lactic acid bacteria and Bacillus licheniformis, thereby achieving the effect of purifying the water. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a picture of the colony morphology of Bacillus subtilis (BSW). DETAILED DESCRIPTION

[0023] The technical solution of the present invention is further explained below in conjunction with the accompanying drawings and specific implementation methods.

[0024] A strain of Bacillus subtilis, Bacillus subtilis B-1158, was deposited in Guangdong Provincial Microbiological Culture Collection Center on December 28, 2021, with the deposit number GDMCC No.62165.

[0025] Specifically, the Bacillus subtilis B-1158 (BSW) in the technical solution was isolated from the sludge at the bottom of the shrimp pond. The total length of the 16S rDNA sequence of the strain is 1390 bp. The 16S rDNA sequence of the BSW sample was compared on NCBI (https: / / b last.ncbi.nlm.nih.gov) and found that the homology between the BSW sample and Bacillus subtilis reached 99.86%. Combined with their colony morphology (such as Figure 1 The strain was identified as Bacillus subtilis and named Bacillus subtilis B-1158. It was deposited in Guangdong Provincial Microbiological Culture Collection Center (abbreviated as: GDMCC, address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Postal Code: 510070) on December 28, 2021, with the deposit number GDMCC No. 62165.

[0026] It is worth noting that the Bacillus subtilis in the technical solution has antibacterial activity and can inhibit Vibrio parahaemolyticus, Vibrio cholerae, Edwardsiella tarda, Aeromonas sobria and Aeromonas hydrophila. Among them, the Bacillus subtilis in the technical solution has excellent antibacterial activity against Vibrio parahaemolyticus. Moreover, the antibacterial ability of the Bacillus subtilis in the solution has the characteristics of high temperature resistance. After the fermentation liquid of BSW is treated at 80°C, the antibacterial activity can be maintained at about 90%, and after being treated at 100°C, the antibacterial activity can still be maintained at about 70%. Based on the use of BSW in the scope of feed or feed additives, the production process of high-temperature granulation may be involved. The high-temperature resistance of the antibacterial activity of BSW makes it more suitable for high-temperature granulation than other strains, and has a wider range of use. At the same time, since BSW is a bacterium separated from the sludge at the bottom of the shrimp pond, it belongs to the indigenous bacteria in the shrimp pond breeding environment and has a stronger adaptability to the breeding ecology, so that BSW and products containing BSW can adapt to the complex environment of the breeding water body and take effect faster. It can be seen that the antibacterial product made of Bacillus subtilis using the technical solution can be used in the aquaculture industry to effectively inhibit the growth and reproduction of Vibrio parahaemolyticus, prevent Vibrio parahaemolyticus from causing harm to marine aquaculture organisms such as shrimp, fish, shellfish and other crustaceans, thereby protecting the safety of marine aquaculture organisms and human body.

[0027] Specifically, the Bacillus subtilis of the present technical solution can form spores under certain conditions. The spores have strong stress resistance and can withstand higher temperatures. Strains with a high spore rate can increase the survival rate of bacteria during the drying process of the powder. The spore rate of the Bacillus subtilis of the present technical solution is above 90%, which makes BSW resistant to high temperatures.

[0028] It is worth pointing out that the method for separating Bacillus subtilis of the present technical solution comprises the following steps:

[0029] (1) An appropriate amount of sample was taken from the sludge at the bottom of the shrimp pond, and after gradient dilution, it was spread on NA solid culture medium and MRS culture medium, respectively. The NA culture medium was cultured at 37°C, and the MRS culture medium was cultured in an anaerobic bag at 37°C. After 16 to 18 hours of culture, the candidate strains were separated according to the colony morphology and obtained.

[0030] (2) After the candidate strains were cultured in NA liquid medium for 24 hours, the supernatant was taken for inhibition zone detection (with Vibrio parahaemolyticus as the indicator bacteria). The candidate strain with the largest inhibition zone diameter was Bacillus subtilis.

[0031] Further description is the use of Bacillus subtilis in the preparation of products for inhibiting Vibrio parahaemolyticus.

[0032] By adding Bacillus subtilis (which can be live bacteria or dried bacterial powder) to the product that inhibits Vibrio parahaemolyticus, Vibrio parahaemolyticus can be effectively inhibited, and Vibrio parahaemolyticus can be prevented from endangering the health of marine aquaculture organisms and human bodies. The product that inhibits Vibrio parahaemolyticus in the technical solution can be an antibacterial agent, a microecological preparation, a feed or a feed additive, etc.

[0033] A bacteriostatic product for aquatic products contains the above-mentioned Bacillus subtilis.

[0034] It is worth noting that the Bacillus subtilis of the present technical solution can inhibit the growth and reproduction of aquatic pathogens such as Vibrio parahaemolyticus, Vibrio cholerae, Edwardsiella tarda, Aeromonas sobriae and Aeromonas hydrophila, and degrade organic matter in aquaculture water; improve the immunity of aquatic animals. By adding Bacillus subtilis to antibacterial products for aquatic use, the activity of Vibrio parahaemolyticus and other aquatic pathogens can be effectively inhibited, thereby ensuring the healthy growth of aquatic animals, achieving the effect of increasing the yield of aquatic animals and protecting human health.

[0035] To further illustrate, the preparation method of the antibacterial product for aquatic products is as follows: Bacillus subtilis and culture medium are prepared into a culture expansion bag to obtain the antibacterial product for aquatic products.

[0036] Specifically, the preparation method of the antibacterial product for aquatic products is as follows: an appropriate amount of culture medium and BSW bacterial powder (obtained after drying) are prepared into a culture expansion bag to obtain the antibacterial product for aquatic products. When using, water is directly added to the antibacterial product for aquatic products, and then aeration and activation are performed for 4 to 5 hours before mixing with feed for feeding.

[0037] Preferably, the culture medium used in the antibacterial product for aquatic products is any one of fermentation medium 1, fermentation medium 2 and fermentation medium 3 in Example 2.

[0038] A water purification product for aquatic products comprises, by weight, 15 to 25 parts of Bacillus subtilis, 15 to 25 parts of lactic acid bacteria, 15 to 25 parts of Bacillus licheniformis and 25 to 55 parts of talc, wherein the Bacillus subtilis is the above-mentioned Bacillus subtilis.

[0039] It is worth noting that BSW can not only play an antibacterial role, but also produce amylase and protease. At the same time, Bacillus licheniformis can produce a variety of digestive enzymes, such as protease, lipase and amylase. The various digestive enzymes that BSW and Bacillus licheniformis can produce can effectively degrade the organic matter in the aquaculture water body, thereby achieving the effect of purifying the water body. At the same time, lactic acid bacteria are added to the aquatic water purification product. Since lactic acid bacteria are anaerobic bacteria, they can adapt to the low-oxygen environment near the bottom of the pond. Through the compounding effect of Bacillus subtilis, lactic acid bacteria and Bacillus licheniformis, a better water purification effect can be achieved. The aquatic water purification product of the technical solution can be used to purify the water body after being poured on water. Experiments have shown that the transparency of the aquaculture water body before the aquatic water purification product is used is about 15 cm, and the transparency is increased to about 20 cm two days after use. It can be seen that the aquatic water purification product of the technical solution can effectively increase the transparency of the aquaculture water body and achieve the effect of purifying the water body. At the same time, the aquatic water purification product of the technical solution can also inhibit a variety of aquatic pathogens and protect aquatic animals.

[0040] Specifically, Bacillus subtilis, lactic acid bacteria and Bacillus licheniformis used in aquatic water purification products are all bacterial powders.

[0041] Preferably, the water purification product for aquaculture includes 20 parts of Bacillus subtilis, 20 parts of lactic acid bacteria, 20 parts of Bacillus licheniformis and 40 parts of talc in parts by mass.

[0042] To further illustrate, the preparation method of the water purification product for aquatic products is as follows: Bacillus subtilis, lactic acid bacteria, Bacillus licheniformis and talcum powder are uniformly mixed to obtain the water purification product for aquatic products.

[0043] It is worth noting that the preparation method of the aquatic water purification product of the present technical solution is simple and has a low preparation cost, and it only requires mixing the raw materials evenly.

[0044] Specifically, when using the aquatic water purification product of the present technical solution, since the Bacillus subtilis, lactic acid bacteria and Bacillus licheniformis used in the aquatic water purification product are all bacterial powders and are in a spore state (dormant body), when using it, it is necessary to add water and brown sugar to the aquatic water purification product for aeration activation for 4 to 5 hours. The activation with water and brown sugar can awaken the Bacillus subtilis, lactic acid bacteria and Bacillus licheniformis from the dormant body, and after activation, it can be used to purify the water body by splashing water.

[0045] Specifically, during activation, the mass ratio of purified water for aquatic products, water, and brown sugar is 1:5:94.

[0046] A liver protection product for aquatic products, comprising, by weight, 45 to 75 parts of compound bacterial powder, 15 to 25 parts of compound traditional Chinese medicine, 5 to 15 parts of antimicrobial peptides and 5 to 15 parts of yeast nucleotides;

[0047] The composite bacterial powder comprises 15 to 25 parts of Bacillus subtilis, 15 to 25 parts of lactic acid bacteria and 15 to 25 parts of Bacillus licheniformis in parts by mass, wherein the Bacillus subtilis in the composite bacterial powder is the above-mentioned Bacillus subtilis.

[0048] To further illustrate, the compound Chinese medicine includes salvia miltiorrhiza, astragalus, wolfberry and schisandra chinensis.

[0049] It is worth noting that the antimicrobial peptides and Bacillus subtilis in the aquatic liver protection product of the present technical scheme both have antibacterial effects, can inhibit harmful bacteria in the intestines of aquatic animals, improve intestinal health, and the digestive system of aquatic animals is healthy, and the load on the liver is relatively reduced; at the same time, lactic acid bacteria can inhibit harmful intestinal bacteria, and the effect is better when used in combination with antimicrobial peptides. Bacillus licheniformis can produce a variety of digestive enzymes (such as proteases, lipases, amylases, etc.), which are helpful for digestion and absorption of aquatic animals. The composite Chinese medicine in the aquatic liver protection product includes salvia miltiorrhiza, astragalus, wolfberry and schisandra chinensis. The composite Chinese medicine has the effects of soothing the liver and regulating qi, clearing the liver and improving eyesight; yeast nucleotides can accelerate the differentiation, growth and repair of intestinal cells, and also have excellent antioxidant effects, which can effectively maintain the normal function of the liver. The aquatic liver protection product of the present technical scheme can achieve the liver protection effect through the synergistic effect of multiple components such as composite bacterial powder, composite Chinese medicine, antimicrobial peptides and yeast nucleotides.

[0050] Specifically, yeast nucleotides can be purchased directly from the market, Shenyu Hubei Haiyi Biotechnology Co., Ltd., model YT500.

[0051] It is further explained that the preparation method of the liver protection product for aquatic products of the present scheme is as follows: according to the formula, the compound bacterial powder, the compound traditional Chinese medicine, the antimicrobial peptide and the yeast nucleotide are evenly mixed to obtain the liver protection product for aquatic products.

[0052] Preferably, the liver protection product for aquatic products includes 60 parts of compound bacterial powder, 20 parts of compound traditional Chinese medicine, 10 parts of antimicrobial peptides and 10 parts of yeast nucleotides in parts by mass.

[0053] Preferably, the mass ratio of Danshen, Astragalus, Lycium barbarum and Schisandra chinensis in the composite traditional Chinese medicine is 1:1:1:1.

[0054] Example 1

[0055] The separation and screening of Bacillus subtilis in the present embodiment are as follows:

[0056] (1) An appropriate amount of sample was taken from the sludge at the bottom of the shrimp pond, and after gradient dilution, it was spread on NA solid culture medium and MRS culture medium, respectively. The NA culture medium was cultured at 37°C, and the MRS culture medium was cultured in an anaerobic bag at 37°C. After 18 hours of culture, the candidate strains were separated according to the colony morphology and obtained.

[0057] (2) After the candidate strains were cultured in NA liquid medium for 24 hours, the supernatant was taken for inhibition zone detection (with Vibrio parahaemolyticus as the indicator bacteria). The candidate strain with the largest inhibition zone diameter was Bacillus subtilis.

[0058] Specifically, the results of the 16S rDNA sequence analysis of Bacillus subtilis in this implementation are as follows: The total length of the 16S rDNA sequence of the strain is 1390 bp. The 16S rDNA sequence of the BSW sample was compared on NCBI (https: / / blast.ncbi.nlm.nih.gov) and found that the homology between the BSW sample and Bacillus subtilis reached 99.86%. Combined with their colony morphology (such as Figure 1 The strain was identified as Bacillus subtilis and named Bacillus subtilis B-1158. It was deposited in Guangdong Provincial Microbiological Culture Collection Center (abbreviated as: GDMCC, address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Postal Code: 510070) on December 28, 2021, with the deposit number GDMCC No.62165.

[0059] Example 2

[0060] Fermentation method of Bacillus subtilis: Pick a single BSW colony and inoculate it into LB medium. After culturing at 37°C and 180rpm for 10 hours, use it as a seed liquid. The seed liquid is inoculated into the three fermentation mediums of formula shown in Table 1 respectively. After inoculation, culture is carried out at 37°C and 180rpm for 24 hours. After the cultivation is completed, microscopic examination is started, and the spore formation time is recorded and the data is detected. The experimental data are shown in Table 2 below.

[0061] Table 1 Formula of fermentation medium

[0062]

[0063]

[0064] Table 2 Experimental data of fermentation using three fermentation media

[0065]

[0066] Specifically, it can be seen from the experimental data in Table 2 that the three fermentation media shown in Table 1 can all be used to ferment Bacillus subtilis to form spores, and the spore rates of Bacillus subtilis fermented using the three fermentation media are all above 90%, especially when fermentation medium 2 is used for fermentation, the spore rate of Bacillus subtilis reaches 99.03%. The spores have strong stress resistance and can withstand higher temperatures. Strains with high spore rates can improve the survival rate of bacteria during the drying process of bacterial powder. The spore rate of BSW in the technical solution is above 90%, which makes BSW have excellent high temperature resistance.

[0067] The high temperature resistance test of Bacillus subtilis is performed as follows: the BSW fermentation liquid obtained by fermentation using the three fermentation media is subjected to high temperature treatment at 80°C and 100°C respectively, and the number of viable bacteria is tested. The test data are shown in Table 3 below:

[0068] Table 3 High temperature resistance test data of Bacillus subtilis

[0069]

[0070] From the test data in Table 3, it can be seen that after the BSW fermentation liquid obtained by fermentation with three fermentation media was subjected to high temperature treatment at 80°C, the bacterial survival rate was greater than 90%; after high temperature treatment at 100°C, the cell survival rate was still maintained at more than 70%. It can be seen that the Bacillus subtilis of this scheme has the characteristics of high temperature resistance, and the cell survival rate of BSW is high after high temperature treatment, so that BSW can still maintain good antibacterial activity after high temperature treatment.

[0071] Example 3

[0072] The antibacterial ability of Bacillus subtilis (BSW) was verified by using the Oxford method for antibacterial test. The specific operation was as follows: the sterilized unsolidified LB solid culture medium was poured into the culture dish, 30 mL per dish. After solidification, 100 μL of the best indicator concentration of the bacterial solution prepared by indicator bacteria (Vibrio parahaemolyticus, Vibrio cholerae, Edwardsiella tarda, Aeromonas sobria and Aeromonas hydrophila) was taken with a pipette, and evenly spread with a sterile coating rod; under sterile conditions, a sterilized Oxford cup (the size of the Oxford cup: inner diameter 6 mm, outer diameter 8 mm, height 10 mm) was placed vertically on the surface of the LB solid culture medium; 100 μL of the sample to be tested (BSW fermentation broth) was added to the Oxford cup, and cultured at 37°C, 180 rpm for 24 h, and the diameter of the inhibition zone was measured. The results are shown in Table 4 below.

[0073] Table 4 Antibacterial effect of Bacillus subtilis (BSW) on pathogenic bacteria

[0074] Indicator bacteria Diameter of inhibition zone (mm) Vibrio parahaemolyticus 27 Vibrio cholerae 21 Edwardsiella tarda 26 Aeromonas sobria 18 Aeromonas hydrophila 20

[0075] As can be seen from Table 4, Bacillus subtilis (BSW) has good antibacterial effects on Vibrio parahaemolyticus, Vibrio cholerae, Edwardsiella tarda, Aeromonas sobria and Aeromonas hydrophila, and has a better antibacterial effect on Vibrio parahaemolyticus, with an inhibition zone diameter of 27 mm.

[0076] Example 4

[0077] The enzyme production capacity of Bacillus subtilis was studied as follows: BSW was inoculated into a culture medium (1.6% yeast extract powder, 2% molasses), and after culturing at 37°C for 24 hours, the supernatant was taken to detect the activity of amylase and neutral protease. The amylase activity was 21.17U / ml, and the neutral protease activity was 459U / ml. From the above test results, it can be seen that the Bacillus subtilis of the present technical solution can produce amylase and protease. Therefore, the use of Bacillus subtilis in aquaculture water can degrade the organic matter in aquaculture water, thereby achieving the effect of purifying the water. At the same time, feeding Bacillus subtilis to aquaculture animals can not only inhibit harmful bacteria, but also help digestion and absorption of aquaculture animals.

[0078] Example 5

[0079] A method for preparing an antibacterial product for aquatic products is as follows: Bacillus subtilis and a culture medium are prepared into a culture expansion package to obtain the antibacterial product for aquatic products, wherein the culture medium is the fermentation medium 2 in Example 2.

[0080] Specifically, when using the antibacterial product for aquaculture of the present embodiment, water is directly added to the antibacterial product for aquaculture to activate it by aeration for 4 to 5 hours, and then the product can be mixed with feed for feeding.

[0081] Example 6

[0082] A water purification product for aquaculture comprises, by weight, 20 parts of Bacillus subtilis, 20 parts of lactic acid bacteria, 20 parts of Bacillus licheniformis and 40 parts of talcum powder, wherein the Bacillus subtilis is Bacillus subtilis B-1158 (BSW for short). The Bacillus subtilis, lactic acid bacteria and Bacillus licheniformis used in the water purification product for aquaculture of this embodiment are all bacterial powders, which can be prepared according to the existing conventional bacterial powder preparation method.

[0083] The preparation method of the water purification product for aquatic products in this embodiment is as follows: Bacillus subtilis, lactic acid bacteria, Bacillus licheniformis and talcum powder are uniformly mixed to obtain the water purification product for aquatic products.

[0084] Specifically, the method of using the water purification product for aquatic products of this embodiment is as follows: add water and brown sugar to the water purification product for aquatic products and perform aeration activation for 5 hours. The water and brown sugar activation awakens Bacillus subtilis, lactic acid bacteria and Bacillus licheniformis from dormant bodies. After activation, the product can be used for purifying water by pouring water. During activation, the mass ratio of the water purification product for aquatic products, water and brown sugar is 1:5:94.

[0085] Specifically, the aquatic water purification product prepared in this embodiment is put into an aquaculture pond. The area of ​​the aquaculture pond is 0.5 acres, and the average water depth is 1.1 meters. The transparency of the aquaculture pond is 15 cm after the aquatic water purification product prepared in this embodiment is put into the aquaculture pond. After two days, the transparency is increased to 21 cm. It can be seen that the aquatic water purification product of this embodiment can effectively increase the transparency of the aquaculture water body and achieve the effect of purifying the water body.

[0086] Example 7

[0087] A liver protection product for aquatic products, comprising, by mass, 60 parts of compound bacterial powder, 20 parts of compound traditional Chinese medicine, 10 parts of antimicrobial peptides and 10 parts of yeast nucleotides; wherein the compound bacterial powder comprises 20 parts of Bacillus subtilis (BSW), 20 parts of lactic acid bacteria and 20 parts of Bacillus licheniformis; the compound traditional Chinese medicine comprises 5 parts of Salvia miltiorrhiza, 5 parts of Astragalus, 5 parts of Lycium barbarum and 5 parts of Schisandra chinensis.

[0088] The preparation method of the liver protection product for aquatic products of this embodiment is as follows: according to the formula, the compound bacterial powder, the compound traditional Chinese medicine, the antimicrobial peptide and the yeast nucleotide are evenly mixed to obtain the liver protection product for aquatic products.

[0089] The technical principle of the present invention is described above in conjunction with specific embodiments. These descriptions are only for explaining the principle of the present invention and cannot be interpreted as limiting the scope of protection of the present invention in any way. Based on the explanations herein, those skilled in the art can associate other specific implementations of the present invention without paying creative labor, and these methods will fall within the scope of protection of the present invention.

Claims

1. A strain of Bacillus subtilis ( Bacillus subtilis ), characterized in that, The bacterium is Bacillus subtilis B-1158, which was deposited in the Guangdong Provincial Microbiological Culture Collection Center on December 28, 2021, with the deposit number GDMCC No.62165.

2. The use of Bacillus subtilis according to claim 1, characterized in that The Bacillus subtilis is used in the preparation of Vibrio parahaemolyticus ( Vibrio parahaemolyticus ) products.

3. An antibacterial product for aquatic products, characterized in that: Contains the Bacillus subtilis according to claim 1.

4. The antibacterial product for aquaculture according to claim 3, characterized in that: The preparation method of the antibacterial product for aquatic products is as follows: Bacillus subtilis and culture medium are prepared into a culture expansion bag to obtain the antibacterial product for aquatic products.

5. A water purification product for aquaculture, characterized in that: By weight, the aquatic water purification product includes 15-25 parts of Bacillus subtilis, 15-25 parts of lactic acid bacteria, and Bacillus licheniformis ( Bacillus licheniformis ) 15-25 parts and talc 25-55 parts, wherein the Bacillus subtilis is the Bacillus subtilis according to claim 1.

6. A liver protection product for aquatic products, characterized in that: By mass, it includes 45-75 parts of compound bacterial powder, 15-25 parts of compound traditional Chinese medicine, 5-15 parts of antimicrobial peptides and 5-15 parts of yeast nucleotides; In terms of mass percentage, the composite bacterial powder comprises 15-25 parts of Bacillus subtilis, 15-25 parts of lactic acid bacteria and 15-25 parts of Bacillus licheniformis, wherein the Bacillus subtilis in the composite bacterial powder is the Bacillus subtilis described in claim 1.

7. The liver protection product for aquatic products according to claim 6, characterized in that: The composite Chinese medicine comprises salvia miltiorrhiza, astragalus, wolfberry and schisandra chinensis.

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