CN112189770A - Antibiotic-free grass carp feed containing fermented mulberry leaves and preparation method thereof - Google Patents
Antibiotic-free grass carp feed containing fermented mulberry leaves and preparation method thereof Download PDFInfo
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Abstract
The invention discloses an antibiotic-free grass carp feed containing fermented mulberry leaves, which comprises the following raw materials in parts by weight: 5-15 parts of fermented mulberry leaf powder, 15-20 parts of soybean meal, 15-20 parts of rapeseed meal, 8-12 parts of cottonseed meal, 1-3 parts of imported fish meal, 16-20 parts of wheat middling, 1-5 parts of corn flour, 8-12 parts of wheat bran, 4-8 parts of rice bran, 0.5-1.5 parts of soybean oil, 1-2 parts of monocalcium phosphate, 0-1.5 parts of bentonite and 1-4 parts of a fish nutrition mixture. The invention can reduce the use of antibiotics and medicines in the process of grass carp culture and overcome various side effects brought by the overuse of medicines to aquaculture; the utilization rate of the bait is improved; improving immunity of grass carp and improving muscle quality.
Description
Technical Field
The invention relates to the technical field of aquaculture feed, in particular to grass carp feed and a preparation method thereof.
Background
Grass carp is one of four Chinese carps cultured in fresh water in China, and is a typical herbivorous fish. The chicken has tender meat quality, rapid growth, coarse feeding resistance and low cultivation cost. At present, in order to pursue the culture benefit, aquaculture farmers excessively use antibiotics, hormones, preservatives, pesticides, chemical fertilizers and other medicines under an intensive high-density culture mode, so that medicine residues are easy to appear on grass carps, and the health of consumers is affected. Meanwhile, a large amount of residual bait generated in the culture process is accumulated at the bottom of the culture pond, so that organic matters are decomposed under the action of anaerobic bacteria to generate a large amount of substances which are toxic to aquaculture animals, and the physicochemical environment and the ecological environment of the culture water body are seriously deteriorated. In order to improve the culture benefit, the problem to be solved is how to replace or reduce the use of antibiotics and medicines in the grass carp culture process and improve the utilization rate of bait.
The mulberry leaves are rich in nutrient substances, are confirmed to be plant leaves with homology of medicine and food by the national ministry of health in 1993, contain 20-36% of crude protein, multiple vitamins and minerals, amino acids, carbohydrates and plant fibers, have high content of essential amino acids for animals, and are one of low-cost protein raw materials of livestock, poultry and aquatic feeds. In addition, the mulberry leaves contain natural active substances such as flavonoid compounds, polysaccharide, sitosterol, isoquercitrin, gamma-amino acid, 1-deoxynojirimycin and the like, have anti-stress effect and can enhance the disease resistance of animal organisms.
Disclosure of Invention
In view of the defects of the existing grass carp feed, the invention provides the antibiotic-free grass carp feed containing the fermented mulberry leaves, which can overcome various side effects of factors such as antibiotics, hormones, preservatives, pesticides and chemical fertilizers on aquaculture and improve the utilization rate of bait, and has the advantages of comprehensive and balanced nutrition, good palatability, easiness in digestion and absorption of grass carps, improvement of the immunity of the grass carps, improvement of muscle quality and the like.
In order to achieve the above purpose, the embodiment of the invention adopts the following technical scheme:
the antibiotic-free grass carp feed containing the fermented mulberry leaves comprises the following raw materials in parts by weight: 5-15 parts of fermented mulberry leaf powder, 15-20 parts of soybean meal, 15-20 parts of rapeseed meal, 8-12 parts of cottonseed meal, 1-3 parts of imported fish meal, 16-20 parts of wheat middling, 1-5 parts of corn flour, 8-12 parts of wheat bran, 4-8 parts of rice bran, 0.5-1.5 parts of soybean oil, 1-2 parts of monocalcium phosphate, 0-1.5 parts of bentonite and 1-4 parts of a fish nutrition mixture.
According to one aspect of the invention, the fermented mulberry leaf powder is prepared by fermenting mulberry leaf powder and auxiliary materials with a microbial starter, drying and crushing, wherein the auxiliary materials comprise glucose, monopotassium phosphate and ammonium sulfate.
According to one aspect of the invention, the mulberry leaf powder, the glucose, the potassium dihydrogen phosphate and the ammonium sulfate are mixed in a mass ratio of 100: 3: 0.25: 1.
according to one aspect of the invention, the microbial starter is a composite microbial agent of acidophilic chlorella and bacillus subtilis, and the ratio of acidophilic chlorella to bacillus subtilis in the microbial starter is 1:1.
according to one aspect of the invention, the antibiotic-free grass carp feed containing fermented mulberry leaves comprises the following raw materials in parts by weight: 10 parts of fermented mulberry leaf powder, 17 parts of soybean meal, 16.2 parts of rapeseed meal, 12 parts of cottonseed meal, 1.5 parts of imported fish meal, 18 parts of wheat middling, 3 parts of corn flour, 10 parts of wheat bran, 6 parts of rice bran, 0.8 part of soybean oil, 1.7 parts of monocalcium phosphate, 0.3 part of bentonite and 3.5 parts of fish nutrition mixture
According to one aspect of the invention, the nutritional blend for fish comprises: vitamin A and vitamin D3Vitamin E, vitamin K3Vitamin B1Vitamin B2Vitamin B6Vitamin B12Vitamin C, calcium pantothenate, nicotinic acid, biotin, folic acid, inositol, choline chloride, betaine, salt, monocalcium phosphate, ferrous sulfate, copper sulfate, manganese sulfate, zinc oxide, calcium iodate, sodium selenite and cobalt sulfate.
A preparation method of antibiotic-free grass carp feed containing fermented mulberry leaves comprises the following steps:
mixing and uniformly stirring the following raw materials in parts by weight: 5-15 parts of fermented mulberry leaf powder, 15-20 parts of soybean meal, 15-20 parts of rapeseed meal, 8-12 parts of cottonseed meal, 1-3 parts of imported fish meal, 16-20 parts of wheat middling, 1-5 parts of corn flour, 8-12 parts of wheat bran, 4-8 parts of rice bran, 0.5-1.5 parts of soybean oil, 1-2 parts of monocalcium phosphate, 0-1.5 parts of bentonite and 1-4 parts of a fish nutrition mixing agent to obtain a mixture;
treating the mixture for 30-45min by steam at the temperature of 100 ℃ and 110 ℃ to obtain a cured mixture;
granulating the cured mixture to obtain granulated feed;
drying the pellet feed to obtain the antibiotic-free grass carp feed containing fermented mulberry leaves.
According to one aspect of the present invention, the preparation method of the fermented mulberry leaf powder comprises the steps of:
crushing the dried mulberry leaves to obtain mulberry leaf powder;
uniformly mixing the mulberry leaf powder with glucose, potassium dihydrogen phosphate and ammonium sulfate, adding a microbial starter, and sealing at normal temperature for 6-10 days to obtain a mulberry leaf fermented product;
drying the mulberry leaf fermented product until the water content is lower than 10%, and crushing at the temperature of-2-5 ℃ to obtain the fermented mulberry leaf powder.
According to one aspect of the invention, the mulberry leaf powder, the glucose, the potassium dihydrogen phosphate and the ammonium sulfate are mixed in a mass ratio of 100: 3: 0.25: 1.
according to one aspect of the invention, the inoculation amount of the microbial starter is 5%, and the solid-to-water ratio of the microbial starter is 1: 1.5.
The implementation of the invention has the advantages that: according to the preparation method of the antibiotic-free grass carp feed containing the fermented mulberry leaves, provided by the invention, the mulberry leaves are fermented, so that the levels of anti-nutritional factors and harmful substances in the mulberry leaf raw materials are reduced, the level of conventional nutritional ingredients is improved, the natural active ingredients of the mulberry leaves are not damaged, and meanwhile, various bioactive factors are added, so that the feeding value and the nutritional value of the mulberry leaves are improved. The fermented mulberry leaf powder is scientifically matched with other feeds and prepared into the granulated feed, so that the feed has comprehensive and balanced nutrition, good palatability, easy digestion and absorption by grass carp, high feed coefficient and less residual feed. Improving immunity of grass carp and improving muscle quality. The antibiotic-free grass carp feed containing the fermented mulberry leaves provided by the invention reduces residual bait and improves the culture environment by improving the immunity of grass carps and increasing the bait coefficient, can completely eliminate the use of antibiotics, reduce the use of hormones, preservatives, pesticides and chemical fertilizers, overcomes various side effects brought to aquaculture by excessive use of the drugs, has the characteristics of no toxic or side effect, no drug resistance, no residue, low cost and the like, and has very high economical efficiency and practicability.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The antibiotic-free grass carp feed containing the fermented mulberry leaves comprises the following raw materials in parts by weight: 5 parts of fermented mulberry leaf powder, 17 parts of soybean meal, 16.5 parts of rapeseed meal, 12 parts of cottonseed meal, 1.5 parts of imported fish meal, 18 parts of wheat middling, 4 parts of corn flour, 11 parts of wheat bran, 8 parts of rice bran, 0.6 part of soybean oil, 1.6 parts of monocalcium phosphate, 1.3 parts of bentonite and 3.5 parts of a fish nutrition mixture.
The nutritional mixture for fish comprises: vitamin A and vitamin D3Vitamin E, vitamin K3Vitamin B1Vitamin B2Vitamin B6Vitamin B12Vitamin C, calcium pantothenate, nicotinic acid, biotin, folic acid, inositol, choline chloride, betaine, salt, monocalcium phosphate, ferrous sulfate, copper sulfate, manganese sulfate, zinc oxide, calcium iodate, sodium selenite and cobalt sulfate.
The antibiotic-free grass carp feed containing fermented mulberry leaves is prepared by the following steps:
crushing the dried mulberry leaves to obtain mulberry leaf powder;
uniformly mixing the mulberry leaf powder with glucose, monopotassium phosphate and ammonium sulfate, adding a microbial starter, and sealing at normal temperature for 8d to obtain a mulberry leaf fermented product, wherein the mass ratio of the mulberry leaf powder to the glucose to the monopotassium phosphate to the ammonium sulfate is 100: 3: 0.25: 1, a microbial starter is a composite microbial inoculum prepared by mixing acidophilic chlorella and bacillus subtilis according to a ratio of 1:1, the inoculation amount of the microbial starter is 5%, and the solid-water ratio of the added microbial starter is 1: 1.5;
drying the mulberry leaf fermented product until the water content is lower than 10%, and crushing at 0 ℃ to obtain the fermented mulberry leaf powder.
Mixing and uniformly stirring the following raw materials: 15 kg of fermented mulberry leaf powder, 51 kg of soybean meal, 49.5 kg of rapeseed meal, 36 kg of cottonseed meal, 4.5 kg of imported fish meal, 54 kg of wheat middling, 12 kg of corn flour, 33 kg of wheat bran, 24 kg of rice bran, 1.8 kg of soybean oil, 4.8 kg of monocalcium phosphate, 3.9 kg of bentonite and 10.5 kg of a fish nutrition mixture to obtain a mixture;
treating the mixture with 110 deg.C steam for 40min to obtain cured mixture;
granulating the cured mixture to obtain granulated feed;
and drying the pellet feed to obtain the antibiotic-free grass carp feed containing the fermented mulberry leaves.
Example 2
The antibiotic-free grass carp feed containing the fermented mulberry leaves comprises the following raw materials in parts by weight: 10 parts of fermented mulberry leaf powder, 17 parts of soybean meal, 16.2 parts of rapeseed meal, 12 parts of cottonseed meal, 1.5 parts of imported fish meal, 18 parts of wheat middling, 3 parts of corn flour, 10 parts of wheat bran, 6 parts of rice bran, 0.8 part of soybean oil, 1.7 parts of monocalcium phosphate, 0.3 part of bentonite and 3.5 parts of a fish nutrition mixture to obtain a mixture.
The nutritional mixture for fish comprises: vitamin A and vitamin D3Vitamin E, vitamin K3Vitamin B1Vitamin B2Vitamin B6Vitamin B12Vitamin C, calcium pantothenate, nicotinic acid, biotin, folic acid, inositol, choline chloride, betaine, salt, monocalcium phosphate, ferrous sulfate, copper sulfate, manganese sulfate, zinc oxide, calcium iodate, sodium selenite and cobalt sulfate.
The antibiotic-free grass carp feed containing fermented mulberry leaves is prepared by the following steps:
crushing the dried mulberry leaves to obtain mulberry leaf powder;
uniformly mixing the mulberry leaf powder with glucose, monopotassium phosphate and ammonium sulfate, adding a microbial starter, and sealing at normal temperature for 8d to obtain a mulberry leaf fermented product, wherein the mass ratio of the mulberry leaf powder to the glucose to the monopotassium phosphate to the ammonium sulfate is 100: 3: 0.25: 1, a microbial starter is a composite microbial inoculum of acidophilic chlorella and bacillus subtilis which are compounded according to a compounding ratio of 1:1, the inoculation amount of the microbial starter is 5%, and the solid-water ratio of the added microbial starter is 1: 1.5;
drying the mulberry leaf fermented product until the water content is lower than 10%, and crushing at 0 ℃ to obtain the fermented mulberry leaf powder.
Mixing and uniformly stirring the following raw materials by weight: 30 kg of fermented mulberry leaf powder, 51 kg of soybean meal, 48.6 kg of rapeseed meal, 36 kg of cottonseed meal, 4.5 kg of imported fish meal, 54 kg of wheat middling, 9 kg of corn flour, 30 kg of wheat bran, 18 kg of rice bran, 2.4 kg of soybean oil, 5.1 kg of monocalcium phosphate, 0.9 kg of bentonite and 10.5 kg of a fish nutrition mixture to obtain a mixture;
treating the mixture with 110 deg.C steam for 40min to obtain cured mixture;
granulating the cured mixture to obtain granulated feed;
and drying the pellet feed to obtain the antibiotic-free grass carp feed containing the fermented mulberry leaves.
Example 3
The antibiotic-free grass carp feed containing the fermented mulberry leaves comprises the following raw materials in parts by weight: 15 parts of fermented mulberry leaf powder, 17 parts of soybean meal, 16.1 parts of rapeseed meal, 12 parts of cottonseed meal, 1.5 parts of imported fish meal, 18 parts of wheat middling, 1 part of corn flour, 9 parts of wheat bran, 4 parts of rice bran, 1.1 parts of soybean oil, 1.8 parts of monocalcium phosphate, 0 part of bentonite and 3.5 parts of a fish nutrition mixture.
The nutritional mixture for fish comprises: vitamin A and vitamin D3Vitamin E, vitamin K3Vitamin B1Vitamin B2Vitamin B6Vitamin B12Vitamin C, calcium pantothenate, nicotinic acid, biotin, folic acid, inositol, choline chloride, betaine, salt, monocalcium phosphate, ferrous sulfate, copper sulfate, manganese sulfate, zinc oxide, calcium iodate, sodium selenite and cobalt sulfate.
The antibiotic-free grass carp feed containing fermented mulberry leaves is prepared by the following steps:
crushing the dried mulberry leaves to obtain mulberry leaf powder;
uniformly mixing the mulberry leaf powder with glucose, monopotassium phosphate and ammonium sulfate, adding a microbial starter, and sealing at normal temperature for 8d to obtain a mulberry leaf fermented product, wherein the mass ratio of the mulberry leaf powder to the glucose to the monopotassium phosphate to the ammonium sulfate is 100: 3: 0.25: 1, a microbial starter is a composite microbial inoculum of acidophilic chlorella and bacillus subtilis which are compounded according to a compounding ratio of 1:1, the inoculation amount of the microbial starter is 5%, and the solid-water ratio of the added microbial starter is 1: 1.5;
drying the mulberry leaf fermented product until the water content is lower than 10%, and crushing at 0 ℃ to obtain the fermented mulberry leaf powder.
Mixing and uniformly stirring the following raw materials by weight: 45 kg of fermented mulberry leaf powder, 51 kg of soybean meal, 48.3 kg of rapeseed meal, 36 kg of cottonseed meal, 4.5 kg of imported fish meal, 54 kg of wheat middling, 3 kg of corn flour, 27 kg of wheat bran, 12 kg of rice bran, 3.3 kg of soybean oil, 5.4 kg of monocalcium phosphate and 10.5 kg of fish nutrient mixture to obtain a mixture;
treating the mixture with 110 deg.C steam for 40min to obtain cured mixture;
granulating the cured mixture to obtain granulated feed;
and drying the pellet feed to obtain the antibiotic-free grass carp feed containing the fermented mulberry leaves.
Example 4
The antibiotic-free grass carp feed containing the fermented mulberry leaves comprises the following raw materials in parts by weight: 6 parts of fermented mulberry leaf powder, 18 parts of soybean meal, 15 parts of rapeseed meal, 11 parts of cottonseed meal, 2 parts of imported fish meal, 19 parts of wheat middling, 5 parts of corn flour, 8 parts of wheat bran, 4.5 parts of rice bran, 0.5 part of soybean oil, 2 parts of monocalcium phosphate, 1.5 parts of bentonite and 2 parts of a fish nutrition mixture.
The nutritional mixture for fish comprises: vitamin A and vitamin D3Vitamin E, vitamin K3Vitamin B1Vitamin B2Vitamin B6Vitamin B12Vitamin C, calcium pantothenate, nicotinic acid, biotin, folic acid, inositol, choline chloride, betaine, salt, monocalcium phosphate, ferrous sulfate, copper sulfate, manganese sulfate, zinc oxide, calcium iodate, sodium selenite and cobalt sulfate.
The antibiotic-free grass carp feed containing fermented mulberry leaves is prepared by the following steps:
crushing the dried mulberry leaves to obtain mulberry leaf powder;
uniformly mixing the mulberry leaf powder with glucose, monopotassium phosphate and ammonium sulfate, adding a microbial starter, and sealing at normal temperature for 6 days to obtain a mulberry leaf fermented product, wherein the mass ratio of the mulberry leaf powder to the glucose to the monopotassium phosphate to the ammonium sulfate is 100: 3: 0.25: 1, a microbial starter is a composite microbial inoculum of acidophilic chlorella and bacillus subtilis which are compounded according to a compounding ratio of 1:1, the inoculation amount of the microbial starter is 5%, and the solid-water ratio of the added microbial starter is 1: 1.5;
drying the mulberry leaf fermented product until the water content is lower than 10%, and crushing at-2 ℃ to obtain the fermented mulberry leaf powder.
Mixing and uniformly stirring the following raw materials by weight: 6 kg of fermented mulberry leaf powder, 18 kg of soybean meal, 15 kg of rapeseed meal, 11 kg of cottonseed meal, 2 kg of imported fish meal, 19 kg of wheat middling, 5 kg of corn flour, 8 kg of wheat bran, 4.5 kg of rice bran, 0.5 kg of soybean oil, 2 kg of monocalcium phosphate, 1.5 kg of bentonite and 2 kg of a fish nutrient mixture to obtain a mixture;
treating the mixture with 105 deg.C steam for 30min to obtain cured mixture;
granulating the cured mixture to obtain granulated feed;
the pellet feed is dried to obtain the antibiotic-free grass carp feed containing the fermented mulberry leaves.
Example 5
The antibiotic-free grass carp feed containing the fermented mulberry leaves comprises the following raw materials in parts by weight: 8 parts of fermented mulberry leaf powder, 15 parts of soybean meal, 20 parts of rapeseed meal, 9 parts of cottonseed meal, 3 parts of imported fish meal, 16 parts of wheat middling, 3.3 parts of corn flour, 9.8 parts of wheat bran, 7 parts of rice bran, 1 part of soybean oil, 1 part of monocalcium phosphate, 0.6 part of bentonite and 4 parts of a fish nutrition mixture.
The nutritional mixture for fish comprises: vitamin A and vitamin D3Vitamin E, vitamin K3Vitamin B1Vitamin B2Vitamin B6Vitamin B12Vitamin C, calcium pantothenate, nicotinic acid, biotin, folic acid, inositol, choline chloride, betaine, salt, calcium biphosphate, ferrous sulfate, copper sulfate, manganese sulfate, zinc oxide and iodic acidCalcium, sodium selenite and cobalt sulfate.
The antibiotic-free grass carp feed containing fermented mulberry leaves is prepared by the following steps:
crushing the dried mulberry leaves to obtain mulberry leaf powder;
uniformly mixing the mulberry leaf powder with glucose, monopotassium phosphate and ammonium sulfate, adding a microbial starter, and sealing at normal temperature for 7d to obtain a mulberry leaf fermented product, wherein the mass ratio of the mulberry leaf powder to the glucose to the monopotassium phosphate to the ammonium sulfate is 100: 3: 0.25: 1, a microbial starter is a composite microbial inoculum of acidophilic chlorella and bacillus subtilis which are compounded according to a compounding ratio of 1:1, the inoculation amount of the microbial starter is 5%, and the solid-water ratio of the added microbial starter is 1: 1.5;
drying the mulberry leaf fermented product until the water content is lower than 10%, and crushing at 2 ℃ to obtain the fermented mulberry leaf powder.
Mixing and uniformly stirring the following raw materials by weight: 8 tons of fermented mulberry leaf powder, 15 tons of bean pulp, 20 tons of rapeseed meal, 9 tons of cottonseed meal, 3 tons of imported fish meal, 16 tons of secondary meal, 3.3 tons of corn meal, 9.8 tons of wheat bran, 7 tons of rice bran, 1 ton of soybean oil, 1 ton of monocalcium phosphate, 0.6 ton of bentonite and 4 tons of fish nutrition mixture are obtained to obtain a mixture;
steam treating the mixture at 108 deg.C for 35min to obtain cured mixture;
granulating the cured mixture to obtain granulated feed;
the pellet feed is dried to obtain the antibiotic-free grass carp feed containing the fermented mulberry leaves.
Example 6
The antibiotic-free grass carp feed containing the fermented mulberry leaves comprises the following raw materials in parts by weight: 13 parts of fermented mulberry leaf powder, 20 parts of soybean meal, 18.3 parts of rapeseed meal, 8 parts of cottonseed meal, 1 part of imported fish meal, 20 parts of wheat middling, 2 parts of corn flour, 12 parts of wheat bran, 5 parts of rice bran, 1.5 parts of soybean oil, 1.3 parts of monocalcium phosphate, 1 part of bentonite and 1 part of fish nutrition mixture.
The nutritional mixture for fish comprises: vitamin A and vitamin D3Vitamin E, vitamin K3Vitamin B1Vitamin B2Vitamin B6Vitamin B12Vitamin C, calcium pantothenate, nicotinic acid, biotin, folic acid, inositol, choline chloride, betaine, salt, monocalcium phosphate, ferrous sulfate, copper sulfate, manganese sulfate, zinc oxide, calcium iodate, sodium selenite and cobalt sulfate.
The antibiotic-free grass carp feed containing fermented mulberry leaves is prepared by the following steps:
crushing the dried mulberry leaves to obtain mulberry leaf powder;
uniformly mixing the mulberry leaf powder with glucose, monopotassium phosphate and ammonium sulfate, adding a microbial starter, and sealing at normal temperature for 10 days to obtain a mulberry leaf fermented product, wherein the mass ratio of the mulberry leaf powder to the glucose to the monopotassium phosphate to the ammonium sulfate is 100: 3: 0.25: 1, a microbial starter is a composite microbial inoculum of acidophilic chlorella and bacillus subtilis which are compounded according to a compounding ratio of 1:1, the inoculation amount of the microbial starter is 5%, and the solid-water ratio of the added microbial starter is 1: 1.5;
drying the mulberry leaf fermented product until the water content is lower than 10%, and crushing at 5 ℃ to obtain the fermented mulberry leaf powder.
Mixing and uniformly stirring the following raw materials by weight: 13 kg of fermented mulberry leaf powder, 20 kg of soybean meal, 18.3 kg of rapeseed meal, 8 kg of cottonseed meal, 1 kg of imported fish meal, 20 kg of wheat middling, 2 kg of corn flour, 12 kg of wheat bran, 5 kg of rice bran, 1.5 kg of soybean oil, 1.3 kg of monocalcium phosphate, 1 kg of bentonite and 1 kg of a fish nutrient mixture to obtain a mixture;
treating the mixture with steam at 100 deg.C for 45min to obtain cured mixture;
granulating the cured mixture to obtain granulated feed;
the pellet feed is dried to obtain the antibiotic-free grass carp feed containing the fermented mulberry leaves.
Example 7
The applicant selected 450 tails of healthy grass carps of the same batch and similar weights (855.55 + -72.35 g), and randomly divided the grass carps into 5 treatment groups, each treatment group having 3 replicates, and each replicate having 30 tails. The control group is fed with basic feed, and the test components comprise a fermented mulberry leaf powder group 1, a fermented mulberry leaf powder group 2, a fermented mulberry leaf powder group 3 and a mulberry leaf powder group. The feed of the fermented mulberry leaf powder group 1 adopts the antibiotic-free grass carp feed containing the fermented mulberry leaves in the embodiment 1, the feed of the fermented mulberry leaf powder group 2 adopts the antibiotic-free grass carp feed containing the fermented mulberry leaves in the embodiment 2, and the feed of the fermented mulberry leaf powder group 3 adopts the antibiotic-free grass carp feed containing the fermented mulberry leaves in the embodiment 3.
Two strains of acidophilic chlorella (Pediococcus acidilactici) and Bacillus subtilis (Bacillus subtilis) used in the fermentation process of the mulberry leaf powder can be obtained by separating and purifying the feed mulberry field soil microorganisms.
Pediococcus acidilactici (Pediococcus acidilactici), also called Pediococcus acidilactici, belongs to Pediococcus and Pediococcus acidilactici strains, and is a gram-positive facultative anaerobe with a spherical thallus and no spores. The acidophilic pediococcus is an animal probiotic, can produce acid (mainly lactic acid), reduce the pH value of the intestinal tract, adjust the flora of the gastrointestinal tract, maintain the micro-ecological balance of the intestinal tract, inhibit pathogenic bacteria of the intestinal tract and enhance the immunity of animals; produce beneficial metabolite, activate the activity of acid protease, participate in the metabolism of organism, prevent the harmful substance from producing. Can be used as intestinal microecological balance agent, feed microbial additive and leaven.
Bacillus subtilis belongs to the genus Bacillus, and has oval to columnar shape, is located in the center or slightly deviated from the center of the strain, and is a gram-positive aerobic bacterium. The bacillus subtilis is a multifunctional microorganism, can inhibit intestinal pathogenic bacteria, promote the growth of beneficial anaerobic bacteria and indirectly inhibit the growth of pathogenic bacteria; can improve the level of immunoglobulin and antibody, and enhance the cellular immunity and humoral immunity of the organism; can synthesize enzymes such as alpha-amylase, protease, lipase, cellulase and the like, can degrade complex organic matters in the vegetable feed, and can play a role together with digestive enzymes in animal bodies in the digestive tract, thereby promoting digestive absorption and improving the utilization rate of the feed. Has strong inhibiting effect on harmful microorganisms such as vibrio, escherichia coli, baculovirus and the like in aquatic products, and effectively prevents diseases such as enteritis, gill rot and the like of aquatic animals. Can secrete a large amount of chitinase, decompose the cell wall of pathogenic fungi to inhibit fungal diseases and toxic and harmful substances in water and purify water; the residual bait, excrement, organic matters and the like in the decomposition pool have a strong function of cleaning garbage particles in water. The bacillus subtilis not only has wider application in feed and aquaculture, but also has wider application in sewage treatment and bio-fertilizer fermentation or fermentation bed manufacture.
The mulberry leaf powder is fermented by the acidophilic pediococcus and the bacillus subtilis, so that the anti-nutritional factors and toxic substances such as tannin, phytoagglutinin and the like in the mulberry leaf raw material can be degraded, macromolecular nutritional substances such as protein, fibers and the like which are difficult to utilize are decomposed to generate micromolecular nutrition which is easy to digest and absorb, a large amount of mycoprotein and beneficial metabolites (lactic acid, lactein, vitamins and the like) are generated at the same time, and the bacterial protein and the beneficial metabolites can play a role in cooperation with certain natural active ingredients in the mulberry leaf, so that the immunity and the disease resistance of an organism are further. The fermented mulberry leaf powder has higher nutritive value and better palatability, is easy to digest and absorb by grass carps, can improve the intestinal health of the grass carps, enhance the immunity of organisms, reduce the occurrence of diseases and improve the quality of the grass carps, and can also play a role in regulating water quality.
The mulberry leaf powder used in the mulberry leaf powder group is not fermented. The feed of the mulberry leaf powder group comprises the following raw materials in parts by weight: 10 parts of mulberry leaf powder, 17 parts of bean pulp, 16.2 parts of rapeseed meal, 12 parts of cottonseed meal, 1.5 parts of imported fish meal, 18 parts of wheat middling, 3 parts of corn flour, 10 parts of wheat bran, 6 parts of rice bran, 0.8 part of soybean oil, 1.7 parts of monocalcium phosphate, 0.3 part of bentonite and 3.5 parts of a fish nutrition mixture.
The feed of the mulberry leaf powder group is prepared by the following steps:
crushing the dried mulberry leaves to obtain mulberry leaf powder;
mixing and uniformly stirring the following raw materials by weight: 30 kg of mulberry leaf powder, 51 kg of bean pulp, 48.6 kg of rapeseed meal, 36 kg of cottonseed meal, 4.5 kg of imported fish meal, 54 kg of wheat middling, 9 kg of corn flour, 30 kg of wheat bran, 18 kg of rice bran, 2.4 kg of soybean oil, 5.1 kg of monocalcium phosphate, 0.9 kg of bentonite and 10.5 kg of a fish nutrient mixture to obtain a mixture;
treating the mixture with 110 deg.C steam for 40min to obtain cured mixture;
granulating the cured mixture to obtain granulated feed;
and drying the granulated feed to obtain the feed of the mulberry leaf powder group.
The feed formulation for each treatment group is shown in table 1.
Table 1 feed formulations (air dry basis) (%)
The test was performed in 15 equally sized, evenly distributed small ponds (mesh separation, 20m by 10m by 1.5m) in the same pond outdoors. Feeding the fish for 1 time in the morning and at night, recording the feeding amount and observing the food intake condition of the fish. The test period is 90 days, wherein the pre-test period is 10 days, and the positive test period is 80 days. When the test is finished, taking blood from muscle tissues and tail veins, and carrying out growth test determination, muscle nutrition index determination and blood biochemical index determination, wherein the specific detection indexes and methods have the following determination results:
1. growth index
The test fish were weighed after fasting for 24h before the test formally started as initial body weight. And (4) fishing out the grass carps in all the ponds by taking the small ponds as a unit to calculate the weight gain rate, the specific growth rate, the bait coefficient and the survival rate after stopping feeding for 24 hours after the culture test is finished. Randomly selecting 3 fish tails with approximate average weight in each small pond, respectively weighing, measuring the body length and the body height, dissecting on an ice tray, separating visceral mass and liver and pancreas, respectively weighing, and calculating the fullness, the viscera-body ratio and the liver-body ratio. The growth performance and the body index are calculated according to the following formula:
weight gain ratio (WGR,%) - (W)t-W0)/W0×100;
Specific growth rate (SGR,%/d) (lnW)t-lnW0)/t×100;
Bait coefficient (FCR) ═ F/(W)t-W0);
Survival rate (SR,%) Nt/N0×100;
Fullness (CF, g/cm)3)=1Wb/L3×100;
Visceral volume ratio (VSI,%) Wv/Wb×100;
Liver body ratio (HIS,%) Wh/Wb×100;
In the formula: n is a radical of0Fish tails (tails) for the start of the experiment; n is a radical oftThe fish tail number (tail) for the end of the test; wtAverage terminal body weight (kg) for test fish; w0Average initial body weight (kg) for the test fish; t is the experimental feeding days (d); f is the average total feed intake (g); wbTo test fish body weight (g); l is the length (cm) of the fish body to be tested; wvFor test fish visceral weight (g); whTo test the weight of the liver and pancreas (g) of fish.
The weight gain rate reflects the relationship between the weight gain of the fish and the initial weight within a period of time, and more intuitively reflects the weight gain condition and the growth condition of animals. The specific growth rate is the ratio of the growth rate to the growth days, and is a common index for measuring the growth condition, and the larger the specific growth rate is, the faster the body weight per day is increased. The bait coefficient is also called 'meat factor', which means the ratio of the bait dosage to the weight gain of fishes in a certain period, and is an index for evaluating the nutritive value and economic effect of the bait, and the smaller the bait coefficient, the higher the bait conversion efficiency and the higher the bait nutritive value. Meanwhile, the bait coefficient and the residual bait accumulation show a negative correlation relationship, namely the higher the bait coefficient is, the less the residual bait accumulation is. The fullness is the ratio of the fish weight to the fish body length, and is an index reflecting the fish fat-lean degree and growth condition. The body ratio of the viscera and the body ratio of the liver are main indexes for judging whether the body function state and the health condition of the test animal are normal or not. The liver ratio of common grass carps is 1-2%, and the excessive liver ratio is not beneficial to the health of fish organisms.
The results of the growth performance and the body index of the grass carp are shown in a table 2. Compared with a control group, the grass carp feed is added with the fermented mulberry leaf powder or the mulberry leaf powder, the weight gain rate and the specific growth rate of the grass carp are increased in different degrees, the feed coefficient of the grass carp is reduced to some extent, and all index data of the fermented mulberry leaf powder group 2 are superior to those of the mulberry leaf powder group. The addition of 10%, 15% fermented mulberry leaf powder and 10% mulberry leaf powder significantly reduces the fullness of grass carp and liver index. This indicates that the body of the grass carp can be adjusted by adding the fermented mulberry leaf powder or the mulberry leaf powder.
TABLE 2 influence of the addition of fermented mulberry leaf powder in different proportions on growth performance and body index
Note: in the same row, the capital letters with different shoulder marks show significant difference ((P <0.01), the lowercase letters with different shoulder marks show significant difference (P <0.05), and the letters with the same shoulder marks or with no marks show insignificant difference (P > 0.05).
2. Index of muscle quality
The meat quality index is as follows: measuring drip loss, weighing the muscle of the back of the fish, suspending the fish in the center of a disposable cup, sealing the fish with a plastic bag, placing the fish in a refrigerator at 4 ℃ for 24 hours, and weighing the fish again; measuring cooked meat rate, weighing back muscle, steaming in water bath for 30min, taking out, air drying for 15min, and weighing; the shearing force is measured by a tenderness meter; muscle hardness was measured using a texture analyzer. General nutritional ingredient indexes of muscles: the moisture content is measured by a constant-temperature drying weight loss method at 105 ℃ (GB 5009.3-2010); the crude protein is determined by adopting a Kjeldahl method (GB 5009.5-2010); the crude fat was determined by Soxhlet extraction (GB 5009.4-2010). The drip loss and the cooked meat rate are calculated according to the following formula:
water drop loss (%) (weight of meat sample before suspension storage-weight of meat sample after suspension storage)/weight of meat sample before suspension storage × 100;
the cooked meat percentage (%) — the weight of the meat sample after cooking/the weight of the meat sample before cooking × 100.
The muscle quality is the comprehensive manifestation of physical and chemical properties of all aspects of the muscle, including muscle tenderness, drip loss, cooked meat rate, texture characteristics and the like. Muscle tenderness is measured by shear force values, with smaller shear forces being better tenderness and vice versa. Drip loss reflects the water holding capacity of muscle protein, the smaller the drip loss, the better the meat quality. The cooked meat rate refers to the weight loss generated by a series of physical and chemical changes of the muscle after being heated at high temperature, and the higher the cooked meat rate is, the smaller the loss is, and the better the meat feeling is. The texture of muscle is an important factor determining its palatability, and is the main sensory and physicochemical index of meat. Hardness is one of important indexes reflecting texture characteristics of fish meat, mainly comes from the content of collagen in a great amount of connective tissues in muscle fiber gaps of the fish meat, the meat is softer when the content of the collagen is low, and the hardness is larger when the content of the collagen is high.
The results of measuring the muscle quality and the index of the conventional nutrient content of grass carp are shown in Table 3. The shearing force of the three fermented mulberry leaf powder groups is lower than that of the mulberry leaf powder group, and the difference is not obvious. The addition of the fermented mulberry leaf powder or the mulberry leaf powder has obvious influence on drip loss of muscles, cooked meat rate and hardness; the drip loss of the muscles of the fermented mulberry leaf powder group 1, the fermented mulberry leaf powder group 2 and the mulberry leaf powder group is extremely lower than that of a control group, wherein the fermented mulberry leaf powder group 2 is the lowest; the cooked meat rate of the fermented mulberry leaf powder group 1, the fermented mulberry leaf powder group 2, the fermented mulberry leaf powder group 3 and the mulberry leaf powder group is higher than that of the control group, and the effect of the fermented mulberry leaf powder group is better than that of the mulberry leaf powder group; the muscle hardness of the fermented mulberry leaf powder group 3 is higher than that of the control group, and the difference between the other groups and the control group is not obvious. Therefore, the quality of the grass carp muscle can be improved by adding the fermented mulberry leaf powder or the mulberry leaf powder, and the effect of the fermented mulberry leaf powder is better. From the aspect of muscle nutrient components, the addition of the fermented mulberry leaf powder or the mulberry leaf powder has no significant influence on the water content and the crude protein content of the grass carp muscle (P is more than 0.05); with the increase of the addition ratio of the fermented mulberry leaf powder or the mulberry leaf powder to 10%, the crude fat content of the muscle of the grass carp is extremely reduced (P is less than 0.01), and the crude fat content can be reduced by 38.24% at most compared with a control group, which shows that the fat deposition of the muscle of the grass carp can be reduced by adding the fermented mulberry leaf powder or the mulberry leaf powder.
Table 3 influence of addition of fermented mulberry leaf powder in different proportions on muscle quality and general nutritional ingredients of grass carp
3. Determination of serum biochemical indicators
Serum lipid metabolism index: total Cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) are measured by an enzyme colorimetric method, and the kit is purchased from Shenzhen Merrill biomedical electronics GmbH. Serum antioxidant index: measuring total superoxide dismutase activity (T-SOD) by xanthine oxidase; the content of Malondialdehyde (MDA) is measured by adopting a TBA method; the total antioxidant capacity (T-AOC) is determined by a colorimetric method. The kits were purchased from Nanjing to build Bio-resources Co.
The physiological and biochemical state in serum can reflect the overall level of the body. Serum Total Cholesterol (TC) and total Triglyceride (TC) are one of the blood lipid indexes, and the higher the cholesterol content is, the higher the triglyceride is, and the higher the probability of hyperlipidemia is. High density lipoprotein cholesterol (HDL-C) is a combination of high density lipoprotein and cholesterol, and transports cholesterol in blood to liver for catabolism to reduce cholesterol deposition, so that high density lipoprotein cholesterol in blood is higher and cholesterol is decomposed faster; generally, the higher the high density lipoprotein cholesterol, the better. Low density lipoprotein cholesterol (LDL-C), which is a combination of low density lipoprotein and cholesterol, has the effect of promoting cholesterol deposition, so that high low density lipoprotein cholesterol in blood, the faster the cholesterol deposition; generally, the lower the low density lipoprotein cholesterol, the better. Superoxide dismutase (T-SOD) is an enzyme which catalyzes superoxide to decompose oxygen and hydrogen peroxide through a disproportionation reaction and has an antioxidation effect, and the higher the activity of superoxide dismutase is, the stronger the antioxidation capability of an organism is. Malondialdehyde (MDA) is the end product of lipid peroxidation and is an indicator of oxidative damage; the malondialdehyde content is high, the lipid oxidation degree is high, and the organism has poor oxidation resistance. The total antioxidant capacity (T-AOC) is an important index for comprehensively judging the strength of the antioxidant capacity of body fluid, particularly cells, a tissue enzyme system and a non-enzyme system, and has obvious significance for determining the health condition of an organism.
The results of lipid metabolism and antioxidant index of grass carp serum are shown in table 4. As can be seen from table 4, the total cholesterol and total triglyceride contents of the serum of the fermented mulberry leaf powder group 1, the fermented mulberry leaf powder group 2, the fermented mulberry leaf powder group 3, or the mulberry leaf powder group were decreased to different degrees, and the fermented mulberry leaf powder group 1, the fermented mulberry leaf powder group 2, and the fermented mulberry leaf powder group 3 were all lower than the mulberry leaf powder group, as compared to the control group. The content of high-density lipoprotein cholesterol is obviously increased by adding the fermented mulberry leaf powder or the mulberry leaf powder, wherein the fermented mulberry leaf powder group 2 is obviously higher than the mulberry leaf powder group. From the oxidation resistance index, the addition of the fermented mulberry leaf powder or the mulberry leaf powder can obviously reduce the serum malondialdehyde and improve the total oxidation resistance and the total superoxide dismutase content. This indicates that the addition of the fermented mulberry leaf powder or mulberry leaf powder can promote lipid metabolism and improve the antioxidant ability.
Table 4 influence of addition of different proportions of fermented mulberry leaf powder on serum lipid metabolism and antioxidant index of grass carp
The test results show that the fermented mulberry leaf powder has no obvious influence on the growth performance of the grass carp, and the addition of the fermented mulberry leaf powder or the mulberry leaf powder can obviously reduce the drip loss and the fat content of muscles, improve the cooked meat rate of the muscles and improve the meat quality; can also obviously improve the oxidation resistance of the grass carp, promote the fat metabolism of the organism, reduce the blood fat and reduce the occurrence of fish diseases.
On the other hand, comparing the fermented mulberry leaf powder group 2 with the mulberry leaf powder group in the transverse direction, most indexes of the fermented mulberry leaf powder group 2 are significantly superior to those of the mulberry leaf powder group. This shows that the effect of the fermented mulberry leaf powder is superior to that of the mulberry leaf powder in all aspects.
The implementation of the invention has the advantages that: according to the preparation method of the antibiotic-free grass carp feed containing the fermented mulberry leaves, the mulberry leaves are fermented, so that the levels of anti-nutritional factors such as tannin, phytoagglutinin and the like and harmful substances in the mulberry leaf raw materials are reduced, the conventional nutritional ingredient level is improved by decomposing macromolecular nutritional substances such as proteins and fibers which are difficult to utilize in the mulberry leaves to generate micromolecular nutrients which are easy to digest and absorb, meanwhile, a large amount of mycoprotein and beneficial metabolites such as lactic acid, lactein and vitamins are produced, and various bioactive factors are increased, so that the feeding value and the nutritional value of the mulberry leaves are improved. The fermented mulberry leaf powder is scientifically matched with other feeds and prepared into the granulated feed, so that the feed has comprehensive and balanced nutrition, good palatability, easy digestion and absorption by grass carp, high feed coefficient and less residual feed. Improving immunity of grass carp and improving muscle quality. The antibiotic-free grass carp feed containing the fermented mulberry leaves provided by the invention reduces residual bait and improves the culture environment by improving the immunity of grass carps and increasing the bait coefficient, can completely eliminate the use of antibiotics, reduce the use of hormones, preservatives, pesticides and chemical fertilizers, overcomes various side effects brought to aquaculture by excessive use of the drugs, has the characteristics of no toxic or side effect, no drug resistance, no residue, low cost and the like, and has very high economical efficiency and practicability.
The above description is only an embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention disclosed herein are intended to be covered by the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (10)
1. The antibiotic-free grass carp feed containing the fermented mulberry leaves is characterized by comprising the following raw materials in parts by weight: 5-15 parts of fermented mulberry leaf powder, 15-20 parts of soybean meal, 15-20 parts of rapeseed meal, 8-12 parts of cottonseed meal, 1-3 parts of imported fish meal, 16-20 parts of wheat middling, 1-5 parts of corn flour, 8-12 parts of wheat bran, 4-8 parts of rice bran, 0.5-1.5 parts of soybean oil, 1-2 parts of monocalcium phosphate, 0-1.5 parts of bentonite and 1-4 parts of a fish nutrition mixture.
2. The antibiotic-free grass carp feed containing fermented mulberry leaves as claimed in claim 1, wherein the fermented mulberry leaf powder is prepared by fermenting mulberry leaf powder and auxiliary materials with a microbial starter, drying and crushing, and the auxiliary materials comprise glucose, potassium dihydrogen phosphate and ammonium sulfate.
3. The antibiotic-free grass carp feed containing fermented mulberry leaves as claimed in claim 2, wherein the mass ratio of the mulberry leaf powder, glucose, potassium dihydrogen phosphate and ammonium sulfate is 100: 3: 0.25: 1.
4. the antibiotic-free grass carp feed containing fermented mulberry leaves as claimed in claim 2, wherein the microbial starter is a composite microbial agent of acidophilic chlorella and bacillus subtilis, and the ratio of acidophilic chlorella to bacillus subtilis in the microbial starter is 1:1.
5. the antibiotic-free grass carp feed containing fermented mulberry leaves as claimed in claim 1, which is characterized by comprising the following raw materials in parts by weight: 10 parts of fermented mulberry leaf powder, 17 parts of soybean meal, 16.2 parts of rapeseed meal, 12 parts of cottonseed meal, 1.5 parts of imported fish meal, 18 parts of wheat middling, 3 parts of corn flour, 10 parts of wheat bran, 6 parts of rice bran, 0.8 part of soybean oil, 1.7 parts of monocalcium phosphate, 0.3 part of bentonite and 3.5 parts of a fish nutrition mixture.
6. The antibiotic-free grass carp feed containing fermented mulberry leaves according to claims 1-5, wherein the nutritional compound for fish comprises: vitamin A and vitamin D3Vitamin E, vitamin K3Vitamin B1Vitamin B2Vitamin B6Vitamin B12Vitamin C, calcium pantothenate, nicotinic acid, biotin, folic acid, inositol, choline chloride, betaine, salt, monocalcium phosphate, ferrous sulfate, copper sulfate, manganese sulfate, zinc oxide, calcium iodate, sodium selenite and cobalt sulfate.
7. A preparation method of antibiotic-free grass carp feed containing fermented mulberry leaves is characterized by comprising the following steps:
mixing and uniformly stirring the following raw materials in parts by weight: 5-15 parts of fermented mulberry leaf powder, 15-20 parts of soybean meal, 15-20 parts of rapeseed meal, 8-12 parts of cottonseed meal, 1-3 parts of imported fish meal, 16-20 parts of wheat middling, 1-5 parts of corn flour, 8-12 parts of wheat bran, 4-8 parts of rice bran, 0.5-1.5 parts of soybean oil, 1-2 parts of monocalcium phosphate, 0-1.5 parts of bentonite and 1-4 parts of a fish nutrition mixing agent to obtain a mixture;
treating the mixture for 30-45min by steam at the temperature of 100 ℃ and 110 ℃ to obtain a cured mixture;
granulating the cured mixture to obtain granulated feed;
drying the pellet feed to obtain the antibiotic-free grass carp feed containing fermented mulberry leaves.
8. The method for preparing antibiotic-free grass carp feed containing fermented mulberry leaves according to claim 7, wherein the method for preparing fermented mulberry leaf powder comprises the following steps:
crushing the dried mulberry leaves to obtain mulberry leaf powder;
uniformly mixing the mulberry leaf powder with glucose, potassium dihydrogen phosphate and ammonium sulfate, adding a microbial starter, and sealing at normal temperature for 6-10 days to obtain a mulberry leaf fermented product;
drying the mulberry leaf fermented product until the water content is lower than 10%, and crushing at the temperature of minus 2-5 ℃ to obtain the fermented mulberry leaf powder.
9. The preparation method of the antibiotic-free grass carp feed containing fermented mulberry leaves as claimed in claim 8, wherein the mass ratio of the mulberry leaf powder, glucose, potassium dihydrogen phosphate and ammonium sulfate is 100: 3: 0.25: 1.
10. the antibiotic-free grass carp feed containing fermented mulberry leaves as claimed in claim 8, wherein the inoculation amount of the microbial starter is 5%, and the solid-to-water ratio of the microbial starter is 1: 1.5.
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