Aquatic fermented feed containing probiotics as well as preparation method and application of aquatic fermented feed

Abstract

The invention provides an aquatic product fermented feed containing probiotics. The fermented feed is prepared from the following raw materials: fish meal, soybean meal, peanut meal, flour, bone meal, fish soluble, soybean oil, phospholipid and the probiotics. According to the invention, the breathing valve fermentation bag is used, so that anaerobic bacteria and probiotics which play a role are cultivated and fermented under an anaerobic or semi-anaerobic condition. Meanwhile, the breeding and growth of aerobic harmful bacteria can be inhibited through anaerobic culture, and the growth of beneficial bacteria such as bacillus, lactic acid bacteria and clostridium butyricum is facilitated; the compounded probiotic group is screened, acid can be stably produced within a short time, the acid production content is high, and after a period of culture, the biological growth curve of the screened probiotic group can reach a balanced state within about 2 days; the compounded probiotic group can generate butyric acid, lactic acid and other substances through metabolism, so that the PH of the digestive tract of the fish body is reduced, the reproduction of beneficial bacteria is promoted, and the growth of harmful bacteria is inhibited.

Description

Technical Field

[0001] The invention relates to the technical field of pesticides, and in particular to an aquatic fermented feed containing probiotics, and a preparation method and application thereof. Background Art

[0002] In recent years, with the rapid development of my country's aquatic industry, the output of aquatic feed has continued to increase, and the problem of feed resource shortage has become increasingly serious. At the same time, intensive farming has led to the frequent occurrence of fish diseases, and the use of antibiotics has led to many environmental and safety problems. Therefore, it is of great significance to find safe and scientific methods to improve the immune function and disease resistance of fish. Studies have reported that fermented feed, as a newly developed green and pollution-free feed resource, can not only alleviate the shortage of feed resources, but also improve the immunity of animals, thus having the potential to replace antibiotics. However, the application effect of fermented feed in aquaculture is affected by many factors.

[0003] Soybean meal is one of the feeds used to supplement protein for many livestock and poultry. The nutrients in it can promote the growth and development of animals. However, the presence of anti-nutritional factors such as protease inhibitors, tannins, lectins, phytic acid, and oligosaccharides inhibits the effectiveness of the feed.

[0004] At present, the use of fermentation strains on the market is still limited to the research of exogenous beneficial bacteria, and the research on endogenous strains is lacking, which limits the development of microbial fermented feed. Secondly, some fermented feeds are mainly fermented naturally. The quality of feed produced by natural fermentation is greatly affected by the strains and is easily contaminated by foreign bacteria. Secondly, the number and activity of fermented bacteria are not stable enough. Secondly, after the fermented feed is ingested, the colonization of the strains is also a problem worth studying.

[0005] In view of the above situation, the present invention provides an aquatic fermented feed containing probiotics and a preparation method and application thereof. Summary of the invention

[0006] The object of the present invention is to provide an aquatic fermented feed containing probiotics. The raw materials for its preparation include, by weight: 50-300 parts of fish meal, 50-300 parts of soybean meal, 50-300 parts of peanut meal, 10-100 parts of flour, 5-50 parts of bone meal, 0-50 parts of fish soluble pulp, 0-50 parts of soybean oil, and 0-50 parts of phospholipids; The probiotics are Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum, and yeast, and the mass ratio is 0.1:0.2:0.1:0.3:0.1.

[0007] The second aspect of the present invention provides a method for preparing the aquatic fermented feed containing probiotics, comprising the following steps: (1) fish meal, soybean meal, peanut meal, flour, bone meal, fish pulp, soybean oil, lecithin, potassium dihydrogen phosphate, potassium dihydrogen sulfate, magnesium sulfate, manganese sulfate, sodium chloride, yeast extract powder and peptone are mixed and pelleted to obtain a basic feed; (2) inoculating probiotics into a fermentation medium at a fermentation temperature of 20-50° C. for 1-5 days; (3) Mix the prepared bacteria with the basic feed in a sealed bag for fermentation. Mix the feed raw materials with the bacteria and place them in a sealed bag equipped with a one-way air valve for fermentation.

[0008] As an embodiment of the present invention, the fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D (+) -glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, lignocellulose 1.0g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0009] As an embodiment of the present invention, the fermentation conditions of step (2) are: the fermentation temperature is 20-50° C., and the fermentation time is 1-5 days.

[0010] As an embodiment of the present invention, the aquatic fermented feed containing probiotics is used for feeding grass carp.

[0011] As an embodiment of the present invention, the aquatic fermented feed containing probiotics is used to protect the intestinal tissue of grass carp.

[0012] By adopting the above technical solution, the present invention has the following beneficial effects: 1. Use a fermentation bag with a breathing valve to cultivate and ferment the anaerobic probiotics under anaerobic or semi-anaerobic conditions. At the same time, anaerobic cultivation can inhibit the reproduction and growth of aerobic harmful bacteria, and is conducive to the growth of beneficial bacteria such as Bacillus, lactic acid bacteria, Clostridium butyricum, etc. 2. The compounded probiotics have been screened and can stably produce acid in a short period of time with a high acid content. After a period of cultivation, the biological growth curve of the screened probiotics can reach a balanced state in about 2 days; 3. The compounded probiotics can produce butyric acid, lactic acid and other substances through metabolism, thereby reducing the pH of the fish's digestive tract, promoting the reproduction of beneficial bacteria, inhibiting the growth of harmful bacteria, improving the activity of digestive enzymes, increasing the digestibility of nutrients and enhancing the immunity of the animal body; it can also produce growth-promoting factors, such as organic acids, B vitamins, K vitamins, etc.

[0013] 4. After fermentation by probiotics, the fermented feed will produce a variety of unsaturated fatty acids and aromatic acids, emit a wine lees smell, accompanied by a special aroma, which will significantly improve the palatability and further promote the fish's feeding.

[0014] 5. Roughage is rich in crude fiber and protein such as cellulose, hemicellulose, pectin, lignin, etc., but these substances are difficult to be digested and absorbed by fish preparations. After ingestion, the digestive burden on the intestine will increase, causing enteritis; while the fermented feed is equivalent to pre-digestion in the fish body in advance, which can not only make up for the amino acids that are easily lacking in conventional feeds, but also enable the rapid conversion of other roughage raw material nutrients, thereby enhancing the digestion, absorption and utilization effect.

[0015] 5. Grass carp is an omnivorous and herbivorous fish with a low demand for high protein. Therefore, the protein content of grass carp feed is generally around 20-30%, which is significantly lower than that of carnivorous or omnivorous fish; it is in line with the characteristics of its digestive system. The energy ratio of grass carp feed will be relatively low because grass carp has a low metabolic level. Excessive energy intake can easily lead to obesity or liver problems. Grass carp is accustomed to swallowing feed from water bodies, so grass carp feed needs to have a certain stability in water to avoid excessive dissolution and cause nutrient loss. DETAILED DESCRIPTION

[0016] The technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0017] Bacillus subtilis was purchased from China Industrial Microbiological Culture Collection Center, Bacillus subtilis CICC®25064.

[0018] Bacillus licheniformis was purchased from China Industrial Microbiological Culture Collection Center, Bacillus licheniformis CICC®10092.

[0019] Lactobacillus plantarum was purchased from China Industrial Microbiological Culture Collection Center, Lactobacillus plantarum CICC®22698.

[0020] Clostridium butyricum was purchased from China Industrial Microbiological Culture Collection Center, Clostridium butyricum CICC®23847.

[0021] Bacillus cereus was purchased from China Industrial Microbiological Culture Collection Center, Bacillus cereus CICC®10185.

[0022] Example 1 The present embodiment provides an aquatic fermented feed containing probiotics, and the raw materials for preparing the feed include: 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish slurry, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone.

[0023] The probiotics are mixed probiotics of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum and yeast in a ratio of 0.1:0.2:0.1:0.3:0.1.

[0024] The preparation method of the aquatic fermented feed containing probiotics is as follows: (1) 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish pulp, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone were mixed and granulated to obtain a basic feed; (2) Probiotics were inoculated into the fermentation medium at a fermentation temperature of 30±5°C for 4 days; the inoculation rate was 12%; The fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D(+)-glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, lignocellulose 1.0g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0025] (3) The prepared strains of bacteria in step (2) are mixed with the basic feed in a sealed bag for fermentation. After the feed raw materials are mixed with the strains of bacteria, they are placed in a sealed bag equipped with a one-way air valve and fermented for 8 days at a temperature of 30±5°C and a water content of 40% at the initial stage of fermentation.

[0026] Wherein, in step (3), the order of magnitude of the bacterial species is 10 8 cfu / g.

[0027] Example 1-1 The present embodiment provides an aquatic fermented feed containing probiotics, and the raw materials for preparing the feed include: 200 parts of fish meal, 150 parts of soybean meal, 200 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish slurry, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone.

[0028] The probiotics are mixed probiotics of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum and yeast in a ratio of 0.1:0.3:0.1:0.2:0.1.

[0029] The preparation method of the aquatic fermented feed containing probiotics is as follows: (1) 200 parts of fish meal, 150 parts of soybean meal, 200 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish pulp, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone are mixed and granulated to obtain a basic feed; (2) Probiotics were inoculated into the fermentation medium at a fermentation temperature of 30±5°C for 4 days; the inoculation rate was 13%; The fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D(+)-glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, lignocellulose 1.5g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0030] (3) The prepared strains of bacteria in step (2) are mixed with the basic feed in a sealed bag for fermentation. After the feed raw materials are mixed with the strains of bacteria, they are placed in a sealed bag equipped with a one-way air valve and fermented for 8 days at a temperature of 30±5°C and a water content of 40% at the initial stage of fermentation.

[0031] Wherein, in step (3), the order of magnitude of the bacterial species is 10 8 cfu / g.

[0032] Example 2 The present embodiment provides an aquatic fermented feed containing probiotics, and the raw materials for preparing the feed include: 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish slurry, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone.

[0033] The probiotics are mixed probiotics with a ratio of 0.1:0.1:0.3:0.1 among Bacillus subtilis, Lactobacillus plantarum, Clostridium butyricum and yeast.

[0034] The preparation method of the aquatic fermented feed containing probiotics is as follows: (1) 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish pulp, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone were mixed and granulated to obtain a basic feed; (2) Probiotics were inoculated into the fermentation medium at a fermentation temperature of 30±5°C for 4 days; the inoculation rate was 12%; The fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D(+)-glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, lignocellulose 1.0g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0035] (3) The prepared strains of bacteria in step (2) are mixed with the basic feed in a sealed bag for fermentation. After the feed raw materials are mixed with the strains of bacteria, they are placed in a sealed bag equipped with a one-way air valve and fermented for 8 days at a temperature of 30±5°C and a water content of 40% at the initial stage of fermentation.

[0036] Wherein, in step (3), the order of magnitude of the bacterial species is 10 8 cfu / g.

[0037] Example 2-1 The present embodiment provides an aquatic fermented feed containing probiotics, and the raw materials for preparing the feed include: 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish slurry, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone.

[0038] The probiotics are mixed probiotics with a ratio of 0.1:0.2:0.1:0.3:0.1 among Bacillus cereus, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum and yeast.

[0039] The preparation method of the aquatic fermented feed containing probiotics is as follows: (1) 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish pulp, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone were mixed and granulated to obtain a basic feed; (2) Probiotics were inoculated into the fermentation medium at a fermentation temperature of 30±5°C for 4 days; the inoculation rate was 12%; The fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D(+)-glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, lignocellulose 1.0g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0040] (3) The prepared strains of bacteria in step (2) are mixed with the basic feed in a sealed bag for fermentation. After the feed raw materials are mixed with the strains of bacteria, they are placed in a sealed bag equipped with a one-way air valve and fermented for 8 days at a temperature of 30±5°C and a water content of 40% at the initial stage of fermentation.

[0041] Wherein, in step (3), the order of magnitude of the bacterial species is 10 8 cfu / g.

[0042] Example 2-2 The present embodiment provides an aquatic fermented feed containing probiotics, and the raw materials for preparing the feed include: 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish slurry, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone.

[0043] The probiotics are mixed probiotics of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum and yeast in a ratio of 0.1:0.1:0.1:0.1:0.1.

[0044] The preparation method of the aquatic fermented feed containing probiotics is as follows: (1) 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish pulp, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone were mixed and granulated to obtain a basic feed; (2) Probiotics were inoculated into the fermentation medium at a fermentation temperature of 30±5°C for 4 days; the inoculation rate was 12%; The fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D(+)-glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, lignocellulose 1.0g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0045] (3) The prepared strains of bacteria in step (2) are mixed with the basic feed in a sealed bag for fermentation. After the feed raw materials are mixed with the strains of bacteria, they are placed in a sealed bag equipped with a one-way air valve and fermented for 8 days at a temperature of 30±5°C and a water content of 40% at the initial stage of fermentation.

[0046] Wherein, in step (3), the order of magnitude of the bacterial species is 10 8 cfu / g.

[0047] Example 2-3 The present embodiment provides an aquatic fermented feed containing probiotics, and the raw materials for preparing the feed include: 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish slurry, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone.

[0048] The probiotics are mixed probiotics of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum and yeast in a ratio of 0.1:0.2:0.1:0.1:0.1.

[0049] The preparation method of the aquatic fermented feed containing probiotics is as follows: (1) 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish pulp, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone were mixed and granulated to obtain a basic feed; (2) Probiotics were inoculated into the fermentation medium at a fermentation temperature of 30±5°C for 4 days; the inoculation rate was 12%; The fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D(+)-glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, lignocellulose 1.0g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0050] (3) The prepared strains of bacteria in step (2) are mixed with the basic feed in a sealed bag for fermentation. After the feed raw materials are mixed with the strains of bacteria, they are placed in a sealed bag equipped with a one-way air valve and fermented for 8 days at a temperature of 30±5°C and a water content of 40% at the initial stage of fermentation.

[0051] Wherein, in step (3), the order of magnitude of the bacterial species is 10 8 cfu / g.

[0052] Embodiment 2-4 The present embodiment provides an aquatic fermented feed containing probiotics, and the raw materials for preparing the feed include: 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish slurry, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone.

[0053] The probiotics are mixed probiotics with a ratio of 0.2:0.2:0.1:0.3:0.1 among Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum and yeast.

[0054] The preparation method of the aquatic fermented feed containing probiotics is as follows: (1) 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish pulp, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone were mixed and granulated to obtain a basic feed; (2) Probiotics were inoculated into the fermentation medium at a fermentation temperature of 30±5°C for 4 days; the inoculation rate was 12%; The fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D(+)-glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, lignocellulose 1.0g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0055] (3) The prepared strains of bacteria in step (2) are mixed with the basic feed in a sealed bag for fermentation. After the feed raw materials are mixed with the strains of bacteria, they are placed in a sealed bag equipped with a one-way air valve and fermented for 8 days at a temperature of 30±5°C and a water content of 40% at the initial stage of fermentation.

[0056] Wherein, in step (3), the order of magnitude of the bacterial species is 10 8 cfu / g.

[0057] Example 3 The present embodiment provides an aquatic fermented feed containing probiotics, and the raw materials for preparing the feed include: 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish slurry, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone.

[0058] The probiotics are mixed probiotics of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum and yeast in a ratio of 0.1:0.2:0.1:0.3:0.1.

[0059] The preparation method of the aquatic fermented feed containing probiotics is as follows: (1) 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish pulp, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone were mixed and granulated to obtain a basic feed; (2) Probiotics were inoculated into the fermentation medium at a fermentation temperature of 30±5°C for 4 days; the inoculation rate was 12%; The fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D(+)-glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0060] (3) The prepared strains of bacteria in step (2) are mixed with the basic feed in a sealed bag for fermentation. After the feed raw materials are mixed with the strains of bacteria, they are placed in a sealed bag equipped with a one-way air valve and fermented for 8 days at a temperature of 30±5°C and a water content of 40% at the initial stage of fermentation.

[0061] Wherein, in step (3), the order of magnitude of the bacterial species is 10 8 cfu / g.

[0062] Example 4 The present embodiment provides an aquatic fermented feed containing probiotics, and the raw materials for preparing the feed include: 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish slurry, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone.

[0063] The probiotics are mixed probiotics of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum and yeast in a ratio of 0.1:0.2:0.1:0.3:0.1.

[0064] The preparation method of the aquatic fermented feed containing probiotics is as follows: (1) 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish pulp, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone were mixed and granulated to obtain a basic feed; (2) Probiotics were inoculated into the fermentation medium at a fermentation temperature of 30±5°C for 4 days; the inoculation rate was 12%; The fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D(+)-glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, lignocellulose 1.0g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0065] (3) The prepared strains of bacteria in step (2) are mixed with the basic feed in a sealed bag for fermentation. After the feed raw materials are mixed with the strains of bacteria, they are placed in a sealed bag equipped with a one-way air valve and fermented for 7 days at a temperature of 30±5°C and a water content of 35% at the initial stage of fermentation.

[0066] Wherein, in step (3), the order of magnitude of the bacterial species is 10 8 cfu / g.

[0067] Example 4-1 The present embodiment provides an aquatic fermented feed containing probiotics, and the raw materials for preparing the feed include: 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish slurry, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone.

[0068] The probiotics are mixed probiotics of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum and yeast in a ratio of 0.1:0.2:0.1:0.3:0.1.

[0069] The preparation method of the aquatic fermented feed containing probiotics is as follows: (1) 200 parts of fish meal, 150 parts of soybean meal, 180 parts of peanut meal, 60 parts of flour, 30 parts of bone meal, 25 parts of fish pulp, 5 parts of soybean oil, 10 parts of phospholipids, 10 parts of potassium dihydrogen phosphate, 5 parts of potassium dihydrogen sulfate, 5 parts of magnesium sulfate, 3 parts of manganese sulfate, 1 part of sodium chloride, 10 parts of yeast extract powder and 40 parts of peptone were mixed and granulated to obtain a basic feed; (2) Probiotics were inoculated into the fermentation medium at a fermentation temperature of 30±5°C for 4 days; the inoculation rate was 12%; The fermentation medium is: peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, D(+)-glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, lignocellulose 1.0g, Tween 80 1.0g, K 2 HPO 4 2.0 g, MgSO 4 7H 2 O 0.2g, MnSO 4 ·H 2 O 0.05 g, CaCO 3 20.0g, agar 15.0g, distilled water 1.0L, natural pH.

[0070] (3) The prepared strains of bacteria in step (2) are mixed with the basic feed in a sealed bag for fermentation. After the feed raw materials are mixed with the strains of bacteria, they are placed in a sealed bag equipped with a one-way air valve and fermented for 8 days at a temperature of 30±5°C and a water content of 45% at the initial stage of fermentation.

[0071] Wherein, in step (3), the order of magnitude of the bacterial species is 10 8 cfu / g.

[0072] Performance Testing Grass carp eat more feed during the high temperature period, and the intestinal load is higher, so this time is chosen to test the effect of fermented feed; Experimental group: the fermented feed of Example 1 to Example 4-1 was used to replace 20% of the conventional feed of grass carp for feeding for 15 consecutive days; control group: conventional feed was fed throughout the whole process; On the 16th day after feeding, the fish were dissected and their intestinal tissues were collected; Test 1: After the fish intestinal tissue was washed with sterile 0.01M PBS solution, it was fixed with tissue fixative and pathological sections were made. The processed sections were placed under a microscope for observation. The experimental results are as follows: Table 1 Test results of test 1

[0073] Test 2: Effects of fermented feed on intestinal oxidative stress and permeability: The CAT (hydrogen peroxide) activity, SOD (superoxide dismutase) activity, MDA content (propylene glycol), ROS content (reactive oxygen species), and GSH activity (glutathione) in the intestinal tissue were detected.

[0074] Table 2 Test results of test 2

[0075] Test 3: Effect of fermented feed on intestinal microbiota: Detect changes in the intestinal microbial community of fish. It was found that compared with the control group, the number of Proteobacteria and Actinobacteria in the intestinal microorganisms of the fish in the example group was significantly reduced by 30%.

[0076] Test 4: During the test, the feeding situation was recorded every day and samples were taken for analysis. It was found that compared with the control group fed only conventional feed, feeding fermented feed containing probiotics could significantly increase the average weight gain of fish by about 150g.

[0077] Table 3 Test results of test 4

[0078] Test 5: Anti-freezing performance test The products prepared in Example 1 to Example 4-1 were placed at 0 degrees Celsius for 24 hours to check their anti-freezing ability.

[0079] Table 4 Test results of test 5

[0080] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. An aquatic fermented feed containing probiotics, characterized in that: The raw materials for its preparation include, by weight: 50-300 parts of fish meal, 50-300 parts of soybean meal, 50-300 parts of peanut meal, 10-100 parts of flour, 5-50 parts of bone meal, 0-50 parts of fish slurry, 0-50 parts of soybean oil, and 0-50 parts of phospholipids; The probiotics are Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Clostridium butyricum, and yeast, and the mass ratio is 0.1:0.2:0.1:0.3:0.

1.

2. A method for preparing aquatic fermented feed containing probiotics according to claim 1, characterized in that: The following steps are involved: (1) fish meal, soybean meal, peanut meal, flour, bone meal, fish pulp, soybean oil, lecithin, potassium dihydrogen phosphate, potassium dihydrogen sulfate, magnesium sulfate, manganese sulfate, sodium chloride, yeast extract powder and peptone are mixed and pelleted to obtain a basic feed; (2) inoculating probiotics into a fermentation medium at a fermentation temperature of 20-50° C. for 1-5 days; (3) Mix the prepared bacteria with the basic feed in a sealed bag for fermentation. Mix the feed raw materials with the bacteria and place them in a sealed bag equipped with a one-way air valve for fermentation.

3. The method for preparing aquatic fermented feed containing probiotics according to claim 2, characterized in that: The fermentation medium comprises: 10.0 g of peptone, 10.0 g of beef extract, 5.0 g of yeast powder, 5.0 g of D(+)-glucose, 5.0 g of sodium acetate, 2.0 g of diammonium citrate, 1.0 g of lignocellulose, 1.0 g of Tween 80, 2.0 g of K2HPO4, 0.2 g of MgSO4·7H2O, 0.05 g of MnSO4·H2O, 20.0 g of CaCO3, 15.0 g of agar, 1.0 L of distilled water, and natural pH.

4. The method for preparing aquatic fermented feed containing probiotics according to claim 2, characterized in that: The fermentation conditions of step (2) are as follows: the fermentation temperature is 20-50° C., and the fermentation time is 1-5 days.

5. The aquatic fermented feed containing probiotics according to claim 1, characterized in that: It is used to feed grass carp.

6. The aquatic fermented feed containing probiotics according to claim 1, characterized in that: It is used to protect the intestinal tissue of grass carp.

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