CN107712462A - A kind of aquatic products high-efficiency active biological fermentation bait and its preparation method and application - Google Patents

A kind of aquatic products high-efficiency active biological fermentation bait and its preparation method and application Download PDF

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CN107712462A
CN107712462A CN201711092012.1A CN201711092012A CN107712462A CN 107712462 A CN107712462 A CN 107712462A CN 201711092012 A CN201711092012 A CN 201711092012A CN 107712462 A CN107712462 A CN 107712462A
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fermentation
medium
aquatic products
bacillus subtilis
culture
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王强
张延涛
王运功
王臣臣
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Guangdong yuzanchen Biotechnology Co.,Ltd.
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Guangzhou Zhi Long Biological Technology Co Ltd
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/189—Enzymes
    • A—HUMAN NECESSITIES
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Abstract

The invention discloses a kind of aquatic products high-efficiency active biological fermentation bait and its preparation method and application.The raw material of aquatic products high-efficiency active biological fermentation bait, by weight percentage, including：Imported fish meal 30~45%, fermented bean dregs 25~45%, enzymolysis protein 0~30%, small peptide 0.5~1.5%, bacillus subtilis 1~5%, bacillus coagulans 2~8%, saccharomycete 0~5%, digestive ferment 0.1~1%, organic acid 0.5~2%, somatomedin 0.5~2%.The aquatic products is with being not added with any antibiotic in high-efficiency active biological fermentation bait, each strain carries out organic assembling, gives full play to the flora advantage of each strain, can not only prevent disease of digestive tract, and fishes and shrimps can be promoted to grow and put on weight, reach the purpose of compound synergic；And can also improve and maintain the gut flora of fishes and shrimps to balance, directly it can be applied in raising fish and shrimp.

Description

A kind of aquatic products high-efficiency active biological fermentation bait and its preparation method and application
Technical field
The present invention relates to technical field of aquaculture feedstuff, more particularly to a kind of aquatic products high-efficiency active biological fermentation bait And its preparation method and application.
Background technology
At present, with the improvement of people's living standards, the quality security problem of food is increasingly subject to country and the weight of the common people Stood in the breach depending on, pork and its internal organs non-staple food traditional as Chinese people.But antibiotic is in aquatic livestock internal organs Residual also generates direct or indirect harm to the health of human body.So reducing or stopping addition of the antibiotic in feed makes With, have become researcher solve pork quality safety problem vital task.In recent years, probiotics, enzyme preparation, The appearance of Chinese herbal and crude drugs preparations and organic acid Substitutes For Antibiotic, played a significantly greater role for solving this problem, but this A little products still do not make perfect compound feed thoroughly break away from the dependence to antibiotic quasi-growth promoter.
19th century microorganism discovery, microbial fermentation enzyme effect and microbiological appearance, fermentation technique is brought into The fast traffic lane of high speed development, obtained in biological energy source conversion, solid-state castoff processing and metabolite production etc. It is widely applied.Numerous document reports, feed are undergone microbial fermentation after effect, and a large amount of lactic acid bacterias, acetic acid are produced in feed It is greatly enhanced with lactic acid concn, significantly reduces feed pH value, feeding fermented feed can improves aquatic livestock Growth performance and the intestinal health for improving aquatic biological.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of aquatic products high-efficiency active biological fermentation bait and its preparation side Method and application.The aquatic products can not only promote fishes and shrimps to grow with high-efficiency active biological fermentation bait, and can also improve and maintain The gut flora balance of fishes and shrimps, directly can be applied in raising fish and shrimp.
In order to solve the above-mentioned technical problem, the technical scheme is that：
In a first aspect, the invention provides a kind of aquatic products high-efficiency active biological fermentation bait, the aquatic products is lived with efficient The raw material of property biofermentation bait, by weight percentage, including：Imported fish meal 30~45%, fermented bean dregs 25~45%, enzyme Solution albumen 0~30%, small peptide 0.5~1.5%, bacillus subtilis 1~5%, bacillus coagulans 2~8%, saccharomycete 0~ 5%th, digestive ferment 0.1~1%, organic acid 0.5~2%, somatomedin 0.5~2%.
Preferably, the aquatic products is not less than 1,000,000,000/gram with high-efficiency active biological fermentation bait active bacteria number.
Preferably, the one kind or several of the digestive ferment in cellulase, amylase, phytase, protease, lipase Kind.
Preferably, the organic acid is selected from fumaric acid, lactic acid, citric acid, malic acid, tartaric acid, sorbic acid, acetic acid, first One or more in acid, propionic acid.
Preferably, it is sub- to be selected from zinc sulfate, sodium selenite, vitamin E, calcium iodate, copper sulphate, sulfuric acid for the somatomedin One or more in iron, manganese sulfate.
Preferably, the bacillus subtilis is to be made using following methods：
(1) inclined-plane culture：The slant medium prepared in following ratios：15~30g/L of glucose, 10~20g/ of peptone L, 0.5~2g/L of sodium chloride, 3~10g/L of yeast extract, agar 15~30g/L, pH 6.5~7.0；Bacillus subtilis strain is connect On kind to slant medium, under the conditions of 25~37 DEG C of temperature, 24~48h is cultivated, obtains bacillus subtilis strain；
(2) first order seed culture：The primary-seed medium prepared in following ratios：10~30g/L of glucose, peptone 10~20g/L, 0.5~2g/L of sodium chloride, yeast extract 3~10g/L, pH 6.5~7.0；By the withered grass gemma obtained by step (1) Bacillus specie is seeded in the primary-seed medium, under the conditions of 25~37 DEG C of temperature, 150~300r/min of rotating speed, culture 24~36h, obtain level liquid seed culture fluid；
(3) secondary seed culture：The secondary seed medium prepared in following ratios：10~30g/L of peptone, beef extract 3~10g/L, 3~7g/L of sodium chloride, dipotassium hydrogen phosphate 0.5~2g/L, pH 6.5~7.0；By the one-level liquid obtained by step (2) Body seed culture fluid is seeded in the secondary seed medium, 180~250r/min of rotating speed, 0.5~1.0vvm of throughput, Under the conditions of 25~37 DEG C of temperature, 24~36h is cultivated, obtains secondary liquid seed culture fluid；
(4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：10~30g/L of peptone, beef extract 3~ 10g/L, 3~10g/L of yeast extract, 0.5~2g/L of potassium dihydrogen phosphate, magnesium sulfate 0.1~1g/L, pH 7.0~7.5；Sent out with liquid The inoculum concentration of ferment culture medium weight 6~10%, the secondary liquid seed culture fluid obtained by step (3) is inoculated in the liquid and sent out In ferment culture medium, under the conditions of 100~200r/min of speed of agitator, 0.5~0.8vvm of throughput, 35~40 DEG C of fermentation temperature, 24~48h of fermented and cultured, obtain fermentation of bacillus subtilis nutrient solution；
(5) prepared by bacterium powder：Fermentation of bacillus subtilis medium centrifugal obtained by step (4) is separated and collects wet withered grass Bacillus body, after mannitol protective agent, Tween 80 are mixed with wet bacillus subtilis thalline, add macropore starch carrier and carry out Absorption, then crushed after low temperature drying and bacillus subtilis powder is made；Wherein, the mannitol protective agent and wet bacillus subtilis The weight ratio of body is 0.15~0.5:1(g/Kg)；The weight ratio of the Tween 80 and wet bacillus subtilis thalline is 5~20:1 (g/Kg)；The weight ratio of the macropore starch carrier and wet bacillus subtilis thalline is 0.2~1:1(Kg/Kg).
Preferably, the bacillus coagulans is to be made using following methods：
(1) inclined-plane culture：The slant medium prepared in following ratios：5~15g/L of peptone, 1~5g/L of beef extract, 2~7g/L of sodium chloride, agar 15~20g/L, pH 7.2~7.4；Bacillus coagulans kind is seeded on slant medium, in Under the conditions of 25~37 DEG C of temperature, 24~48h is cultivated, obtains bacillus coagulans kind；
(2) first order seed culture：The primary-seed medium prepared in following ratios：5~15g/L of peptone, beef extract 1 ~5g/L, sodium chloride 2~7g/L, pH 7.2~7.4；Bacillus coagulans kind obtained by step (1) is seeded to the one-level In seed culture medium, under the conditions of 25~37 DEG C of temperature, 150~300r/min of rotating speed, 18~30h is cultivated, obtains level liquid Seed culture fluid；
(3) secondary seed culture：The secondary seed medium prepared in following ratios：5~15g/L of peptone, beef extract 1 ~5g/L, 2~7g/L of sodium chloride, pH7.2~7.4；Level liquid seed culture fluid obtained by step (2) is seeded to described two In level seed culture medium, under the conditions of 180~250r/min of rotating speed, 25~37 DEG C of temperature, 18~30h is cultivated, obtains two level liquid Body seed culture fluid；
(4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：20~30g/L of glucose, peptone 10~ 20g/L, 5~15g/L of dusty yeast, 0.5~2g/L of potassium dihydrogen phosphate, magnesium sulfate 0.1~1g/L, pH 7.0~7.5；Sent out with liquid The inoculum concentration of ferment culture medium weight 6~10%, the secondary liquid seed culture fluid obtained by step (3) is inoculated in the liquid and sent out In ferment culture medium, under the conditions of 100~200r/min of speed of agitator, 35~50 DEG C of fermentation temperature, 20~30h of fermented and cultured, obtain To bacillus coagulans fermentation culture；
(5) prepared by bacterium powder：Bacillus coagulans fermentation culture obtained by step (4) is centrifuged and collects wet condensation Bacillus body, after mannitol protective agent, Tween 80 are mixed with wet bacillus coagulans body, add macropore starch carrier and carry out Absorption, then crushed after low temperature drying and bacillus coagulans powder is made；Wherein, the mannitol protective agent and wet bacillus coagulans The weight ratio of body is 0.15~0.5:1(g/Kg)；The weight ratio of the Tween 80 and wet bacillus coagulans body is 5~20:1 (g/Kg)；The weight ratio of the macropore starch carrier and wet bacillus coagulans body is 0.2~1:1(Kg/Kg).
Preferably, the saccharomycete is to be made using following methods：
(1) inclined-plane culture：Barms is inoculated in sugared content 5~20%, pH 4.0~7.0 brewer's wort inclined-plane culture On base, under the conditions of 25~36 DEG C, 25~50h is cultivated, obtains inclined-plane barms；
(2) level liquid seed culture：To be inoculated in after the dilution of inclined-plane barms obtained by step (1) sugared content 5~ 20%th, in pH 4.0~7.0 malt extract medium, under the conditions of 25~36 DEG C, 25~50h is cultivated, obtains level liquid kind Sub- nutrient solution；
(3) secondary liquid seed culture：By the level liquid seed culture fluid obtained by step (2) be inoculated in sugared content 5~ 20%th, in pH 4.0~7.0 malt extract medium, under the conditions of 25~36 DEG C, 25~50h is cultivated, obtains secondary liquid kind Sub- nutrient solution；
(4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：10~30g/L of glucose, peptone 5~ 15g/L, 1~5g/L of potassium dihydrogen phosphate, 0.1~0.5g/L of magnesium sulfate, 0.3~0.6g/L of ferrous sulfate, manganese sulfate 0.3~ 0.6g/L；With the inoculum concentration of liquid fermentation medium weight 6~10%, by the secondary liquid seed culture fluid obtained by step (3) It is inoculated in liquid fermentation medium, in 28~32 DEG C of cultivation temperature, pH 5.5~6.5,0.03~0.06MPa of pressure, rotating speed Under conditions of 200~350r/min, 25~50h of fermented and cultured, saccharomycetes to make fermentation nutrient solution is obtained；
(5) prepared by bacterium powder：Saccharomycetes to make fermentation medium centrifugal obtained by step (4) is separated and collects wet yeast thalline, After mannitol protective agent, Tween 80 are mixed with wet yeast thalline, add macropore starch carrier and adsorbed, then after low temperature drying Crush and yeast bacterium powder is made；Wherein, the weight ratio of the mannitol protective agent and wet yeast thalline is 0.15~0.5:1(g/ Kg)；The weight ratio of the Tween 80 and wet yeast thalline is 5~20:1(g/Kg)；The macropore starch carrier and wet yeast bacterium The weight ratio of body is 0.2~1:1(Kg/Kg).
Second aspect, present invention also offers a kind of aquatic products prepared as described in relation to the first aspect to be fermented with high-efficiency active biological The method of bait, comprises the following steps：
The bacillus subtilis, bacillus coagulans, saccharomycete are mixed into composite bacteria agent, then the compound bacteria by gained Agent, imported fish meal, fermented bean dregs, enzymolysis protein, small peptide, digestive ferment, organic acid, somatomedin mixing, fermented obtained water High-efficiency active biological fermentation bait is used in production.
The third aspect, present invention also offers a kind of aquatic products as described in relation to the first aspect high-efficiency active biological fermentation bait Application in feed is prepared.
Compared with prior art, the beneficial effects of the present invention are：
(1) bacillus subtilis in compound bacteria of the present invention, bacillus coagulans, saccharomycete be respectively provided with the speed of growth it is fast, The strong characteristic of resistance to inverse environment capacity, it can effectively suppress the life of a large amount of miscellaneous bacterias, harmful bacteria contained in feeding system, feedstuff Long breeding, ensures the good fermentation of fermentation solid-state feed；The growth of saccharomycete can decompose synthesizes micro- life using carbohydrate Thing albumen, the inorganic nitrogen-sourced yield that can improve mycoprotein such as a small amount of urea is added during fermentation；Each strain carries out organic assembling, The flora advantage of each strain is given full play to, disease of digestive tract can not only be prevented, and fishes and shrimps can be promoted to grow and increase body Weight, reach the purpose of compound synergic.
(2) aquatic products of the invention is simple with the preparation technology of high-efficiency active biological fermentation bait, fermentation process and fermentation Technological parameter is simple and convenient all by applicant's experiment, optimization, is easy to all kinds of raisers, production Feed Enterprise operation；And prepare Solid state fermentation feed efficiency it is notable, can not only improve efficiency of feed utilization, promote fishes and shrimps growth, and can also improve and maintain Fishes and shrimps gut flora balances, and improves fishes and shrimps immunity and premunition, improves aquaculture economic benefit.
(3) preparation process of aquatic products high-efficiency active biological fermentation bait of the invention and adding for final products antibiotic-free Add, it is green safe, directly it can be applied in raising fish and shrimp.
Brief description of the drawings
The survival rate statistical chart for the Penaeus Vannmei that Fig. 1 is provided by effect example 1 of the present invention；
The output value statistical chart for the Penaeus Vannmei that Fig. 2 is provided by effect example 1 of the present invention；
The survival rate statistical chart for the Tilapia mossambica that Fig. 3 is provided by effect example 2 of the present invention；
The output value statistical chart for the Tilapia mossambica that Fig. 4 is provided by effect example 2 of the present invention.
Embodiment
The embodiment of the present invention is described further below in conjunction with the accompanying drawings.Herein it should be noted that for The explanation of these embodiments is used to help understand the present invention, but does not form limitation of the invention.It is in addition, disclosed below As long as each embodiment of the invention in involved technical characteristic do not form conflict can each other and be mutually combined.
In a first aspect, the invention provides a kind of aquatic products high-efficiency active biological fermentation bait, the aquatic products is lived with efficient The raw material of property biofermentation bait, by weight percentage, including：Imported fish meal 30~45%, fermented bean dregs 25~45%, enzyme Solution albumen 0~30%, small peptide 0.5~1.5%, bacillus subtilis 1~5%, bacillus coagulans 2~8%, saccharomycete 0~ 5%th, digestive ferment 0.1~1%, organic acid 0.5~2%, somatomedin 0.5~2%.
In embodiments of the present invention, the aquatic products is not less than 1,000,000,000 with high-efficiency active biological fermentation bait active bacteria number Individual/gram.
In embodiments of the present invention, the digestive ferment is selected from cellulase, amylase, phytase, protease, lipase In one or more.
In a preferred embodiment of this invention, the digestive ferment includes cellulase, phytase, protease, wherein, it is described Cellulase, phytase, protease weight ratio are 2~5:2~4:1.
In embodiments of the present invention, the organic acid is selected from fumaric acid, lactic acid, citric acid, malic acid, tartaric acid, mountain One or more in pears acid, acetic acid, formic acid, propionic acid.
In embodiments of the present invention, the somatomedin be selected from zinc sulfate, sodium selenite, vitamin E, calcium iodate, One or more in copper sulphate, ferrous sulfate, manganese sulfate.
In embodiments of the present invention, the raw material of aquatic products high-efficiency active biological fermentation bait, by weight percentage Meter, including：Imported fish meal 45%, fermented bean dregs 42%, small peptide 0.5%, bacillus subtilis 5%, bacillus coagulans 2%, Saccharomycete 2%, digestive ferment 0.5%, organic acid 2%, somatomedin 1%.
In embodiments of the present invention, the raw material of aquatic products high-efficiency active biological fermentation bait, by weight percentage Meter, including：Imported fish meal 42%, fermented bean dregs 45%, small peptide 1%, bacillus subtilis 2%, bacillus coagulans 3%, ferment Female bacterium 3%, digestive ferment 1%, organic acid 1.5%, somatomedin 1.5%.
In embodiments of the present invention, the raw material of aquatic products high-efficiency active biological fermentation bait, by weight percentage Meter, including：Imported fish meal 45%, fermented bean dregs 36%, small peptide 1.5%, bacillus subtilis 3%, bacillus coagulans 5%, Saccharomycete 5%, digestive ferment 0.5%, organic acid 2%, somatomedin 2%.
In embodiments of the present invention, the raw material of aquatic products high-efficiency active biological fermentation bait, by weight percentage Meter, including：Imported fish meal 30%, fermented bean dregs 30%, enzymolysis protein 30%, small peptide 0.5%, bacillus subtilis 1%, condensation Bacillus 6%, digestive ferment 0.5%, organic acid 1%, somatomedin 1%.
In embodiments of the present invention, the raw material of aquatic products high-efficiency active biological fermentation bait, by weight percentage Meter, including：Imported fish meal 30%, fermented bean dregs 40%, enzymolysis protein 20%, small peptide 1%, bacillus subtilis 2%, condensation bud Spore bacillus 4%, digestive ferment 0.5%, organic acid 1%, somatomedin 1.5%.
In embodiments of the present invention, the raw material of aquatic products high-efficiency active biological fermentation bait, by weight percentage Meter, including：Imported fish meal 35%, fermented bean dregs 45%, enzymolysis protein 12%, small peptide 1%, bacillus subtilis 3%, condensation bud Spore bacillus 2.5%, digestive ferment 0.5%, organic acid 0.5%, somatomedin 0.5%.
In embodiments of the present invention, the bacillus subtilis is to be made using following methods：
(1) inclined-plane culture：The slant medium prepared in following ratios：15~30g/L of glucose, 10~20g/ of peptone L, 0.5~2g/L of sodium chloride, 3~10g/L of yeast extract, agar 15~30g/L, pH 6.5~7.0；Bacillus subtilis strain is connect On kind to slant medium, under the conditions of 25~37 DEG C of temperature, 24~48h is cultivated, obtains bacillus subtilis strain；
(2) first order seed culture：The primary-seed medium prepared in following ratios：10~30g/L of glucose, peptone 10~20g/L, 0.5~2g/L of sodium chloride, yeast extract 3~10g/L, pH 6.5~7.0；By the withered grass gemma obtained by step (1) Bacillus specie is seeded in the primary-seed medium, under the conditions of 25~37 DEG C of temperature, 150~300r/min of rotating speed, culture 24~36h, obtain level liquid seed culture fluid；
(3) secondary seed culture：The secondary seed medium prepared in following ratios：10~30g/L of peptone, beef extract 3~10g/L, 3~7g/L of sodium chloride, dipotassium hydrogen phosphate 0.5~2g/L, pH 6.5~7.0；By the one-level liquid obtained by step (2) Body seed culture fluid is seeded in the secondary seed medium, 180~250r/min of rotating speed, 0.5~1.0vvm of throughput, Under the conditions of 25~37 DEG C of temperature, 24~36h is cultivated, obtains secondary liquid seed culture fluid；
(4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：10~30g/L of peptone, beef extract 3~ 10g/L, 3~10g/L of yeast extract, 0.5~2g/L of potassium dihydrogen phosphate, 0.1~1g/L of magnesium sulfate, pH7.0~7.5；Sent out with liquid The inoculum concentration of ferment culture medium weight 6~10%, the secondary liquid seed culture fluid obtained by step (3) is inoculated in the liquid and sent out In ferment culture medium, under the conditions of 100~200r/min of speed of agitator, 0.5~0.8vvm of throughput, 35~40 DEG C of fermentation temperature, 24~48h of fermented and cultured, obtain fermentation of bacillus subtilis nutrient solution；
(5) prepared by bacterium powder：Fermentation of bacillus subtilis medium centrifugal obtained by step (4) is separated and collects wet withered grass Bacillus body, after mannitol protective agent, Tween 80 are mixed with wet bacillus subtilis thalline, add macropore starch carrier and carry out Absorption, then crushed after low temperature drying and bacillus subtilis powder is made；Wherein, the mannitol protective agent and wet bacillus subtilis The weight ratio of body is 0.15~0.5:1(g/Kg)；The weight ratio of the Tween 80 and wet bacillus subtilis thalline is 5~20:1 (g/Kg)；The weight ratio of the macropore starch carrier and wet bacillus subtilis thalline is 0.2~1:1(Kg/Kg).
It is understood that bacillus subtilis of the present invention is a kind of gram-positive bacterium, bacillus subtilis Caused withered grass element, polymyxins, nystatin, gramicidins isoreactivity material are to pathogenic bacteria or endogenous during bacteria growing The conditioned pathogen of infection has obvious inhibitory action；After fish, shrimp intake bacillus subtilis, bacillus subtilis consumes rapidly Free oxygen in environment, enteron aisle hypoxemia is caused, promoted beneficial to anaerobism bacteria growing, and produce the organic acids such as lactic acid, reduce enteron aisle PH value, suppress other pathogenic bacteria growings indirectly；Stimulating animal immune organ sense organ grows, and activation T, bone-marrow-derived lymphocyte, carries High immunoglobulinlg and antibody level, strengthen cellular immunity and humoral immune function, improve herd immunity；Bacillus subtilis Thalline itself synthesis alpha-amylase, protease, lipase, the enzyme such as cellulase, in alimentary canal with the digestion in animal body Enzyme together plays a role；Vitamin B can be synthesized1、B2、B6, a variety of B family vitamins such as nicotinic acid, improve fish, disturb in shrimp body The activity of element and macrophage.
In embodiments of the present invention, the bacillus coagulans is to be made using following methods：
(1) inclined-plane culture：The slant medium prepared in following ratios：5~15g/L of peptone, 1~5g/L of beef extract, 2~7g/L of sodium chloride, agar 15~20g/L, pH 7.2~7.4；Bacillus coagulans kind is seeded on slant medium, in Under the conditions of 25~37 DEG C of temperature, 24~48h is cultivated, obtains bacillus coagulans kind；
(2) first order seed culture：The primary-seed medium prepared in following ratios：5~15g/L of peptone, beef extract 1 ~5g/L, 2~7g/L of sodium chloride, pH7.2~7.4；Bacillus coagulans kind obtained by step (1) is seeded to the one-level kind In sub- culture medium, under the conditions of 25~37 DEG C of temperature, 150~300r/min of rotating speed, 18~30h is cultivated, obtains level liquid kind Sub- nutrient solution；
(3) secondary seed culture：The secondary seed medium prepared in following ratios：5~15g/L of peptone, beef extract 1 ~5g/L, 2~7g/L of sodium chloride, pH7.2~7.4；Level liquid seed culture fluid obtained by step (2) is seeded to described two In level seed culture medium, under the conditions of 180~250r/min of rotating speed, 25~37 DEG C of temperature, 18~30h is cultivated, obtains two level liquid Body seed culture fluid；
(4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：20~30g/L of glucose, peptone 10~ 20g/L, 5~15g/L of dusty yeast, 0.5~2g/L of potassium dihydrogen phosphate, magnesium sulfate 0.1~1g/L, pH 7.0~7.5；Sent out with liquid The inoculum concentration of ferment culture medium weight 6~10%, the secondary liquid seed culture fluid obtained by step (3) is inoculated in the liquid and sent out In ferment culture medium, under the conditions of 100~200r/min of speed of agitator, 35~50 DEG C of fermentation temperature, 20~30h of fermented and cultured, obtain To bacillus coagulans fermentation culture；
(5) prepared by bacterium powder：Bacillus coagulans fermentation culture obtained by step (4) is centrifuged and collects wet condensation Bacillus body, after mannitol protective agent, Tween 80 are mixed with wet bacillus coagulans body, add macropore starch carrier and carry out Absorption, then crushed after low temperature drying and bacillus coagulans powder is made；Wherein, the mannitol protective agent and wet bacillus coagulans The weight ratio of body is 0.15~0.5:1(g/Kg)；The weight ratio of the Tween 80 and wet bacillus coagulans body is 5~20:1 (g/Kg)；The weight ratio of the macropore starch carrier and wet bacillus coagulans body is 0.2~1:1(Kg/Kg).
It is understood that bacillus coagulans (Bacillus coagulans) of the present invention, Gram-positive Bacterium, hard (or thick) wall bacterium door is not only belonged to, and belong to enteron aisle bacillus coagulans, also known as " have spore to condense gemma bar Bacterium ".Be through U.S. FDA approval one kind " generally believing safety " bacillus class bacillus coagulans, bacillus coagulans fish, In the enteron aisle of shrimp, it can ferment and produce a large amount of antibacterial coaguin, lactic acid, amino acid, vitamin and a variety of digestive ferments.Condense gemma bar The functional characteristics of bacterium is as follows：(1) effectively suppress the growth of harmful bacteria, improve intestine microenvironment, promote intestinal growth, enhancing Function of intestinal canal；(2) feed quality is improved, promotes digesting and assimilating for feed, reduces feedstuff-meat ratio；(3) be effectively improved weanling pig by In stress be with dysbiosis diarrhea caused by allergic reaction；(4) it is without any side effects, it is safe to use, and environment can be reduced In ammonia, the pernicious gas such as hydrogen sulfide, improve breeding environment.
In embodiments of the present invention, the saccharomycete is to be made using following methods：
(1) inclined-plane culture：Barms is inoculated in sugared content 5~20%, pH 4.0~7.0 brewer's wort inclined-plane culture On base, under the conditions of 25~36 DEG C, 25~50h is cultivated, obtains inclined-plane barms；
(2) level liquid seed culture：To be inoculated in after the dilution of inclined-plane barms obtained by step (1) sugared content 5~ 20%th, in pH 4.0~7.0 malt extract medium, under the conditions of 25~36 DEG C, 25~50h is cultivated, obtains level liquid kind Sub- nutrient solution；
(3) secondary liquid seed culture：By the level liquid seed culture fluid obtained by step (2) be inoculated in sugared content 5~ 20%th, in pH 4.0~7.0 malt extract medium, under the conditions of 25~36 DEG C, 25~50h is cultivated, obtains secondary liquid kind Sub- nutrient solution；
(4) fermentation tank culture：Liquid fermentation medium is prepared in following ratios：10~30g/L of glucose, peptone 5~ 15g/L, 1~5g/L of potassium dihydrogen phosphate, 0.1~0.5g/L of magnesium sulfate, 0.3~0.6g/L of ferrous sulfate, manganese sulfate 0.3~ 0.6g/L；With the inoculum concentration of liquid fermentation medium weight 6~10%, by the secondary liquid seed culture fluid obtained by step (3) It is inoculated in liquid fermentation medium, in 28~32 DEG C of cultivation temperature, pH 5.5~6.5,0.03~0.06MPa of pressure, rotating speed Under the conditions of 200~350r/min, 25~50h is cultivated, obtains saccharomycetes to make fermentation nutrient solution；
(5) prepared by bacterium powder：Saccharomycetes to make fermentation medium centrifugal obtained by step (4) is separated and collects wet yeast thalline, After mannitol protective agent, Tween 80 are mixed with wet yeast thalline, add macropore starch carrier and adsorbed, then after low temperature drying Crush and yeast bacterium powder is made；Wherein, the weight ratio of the mannitol protective agent and wet yeast thalline is 0.15~0.5:1(g/ Kg)；The weight ratio of the Tween 80 and wet yeast thalline is 5~20:1(g/Kg)；The macropore starch carrier and wet yeast bacterium The weight ratio of body is 0.2~1:1(Kg/Kg).
Saccharomycete of the present invention has good stomach juice-resistant, bile tolerance, high temperature resistant, tolerance common antibiotics etc. degeneration-resistant Prebiotic function, the growth of saccharomycete such as performance and production acid, producing enzyme, suppression pathogen can decompose
Add that a small amount of urea etc. is inorganic nitrogen-sourced can to improve thalline using carbohydrate synthetic animalcule albumen, during fermentation The yield of albumen.
Second aspect, present invention also offers a kind of aquatic products prepared as described in relation to the first aspect to be fermented with high-efficiency active biological The method of bait, comprises the following steps：
The bacillus subtilis, bacillus coagulans, saccharomycete are mixed into composite bacteria agent, then the compound bacteria by gained Agent, imported fish meal, fermented bean dregs, enzymolysis protein, small peptide, digestive ferment, organic acid, somatomedin mixing, fermented obtained water High-efficiency active biological fermentation bait is used in production.
In embodiments of the present invention, the condition of the fermentation is：Temperature is 25~35 DEG C, and rotating speed is 150~400r/ Min, incubation time are 3~10 days.
The third aspect, present invention also offers a kind of aquatic products as described in relation to the first aspect high-efficiency active biological fermentation bait Application in feed is prepared.
In embodiments of the present invention, added with aquatic products high-efficiency active biological as described in relation to the first aspect in the feed Fermentation bait, the aquatic products is with 0.05~0.5 times that the addition of high-efficiency active biological fermentation bait is feed gross weight.
Embodiment 1
A kind of preparation method the embodiments of the invention provide aquatic products with high-efficiency active biological fermentation bait, including following step Suddenly：
(1) bacillus subtilis is to be made using following methods：
Inclined-plane culture：The slant medium prepared in following ratios：Glucose 20g/L, peptone 15g/L, sodium chloride 1g/ L, yeast extract 5g/L, agar 20g/L, pH 6.5；Bacillus subtilis strain is seeded on slant medium, in 28 DEG C of bars of temperature Under part, 30h is cultivated, obtains bacillus subtilis strain；
First order seed culture：The primary-seed medium prepared in following ratios：Glucose 20g/L, peptone 15g/L, Sodium chloride 1g/L, yeast extract 5g/L, pH 6.5；The bacillus subtilis strain of gained is seeded to the primary-seed medium In, under the conditions of 30 DEG C of temperature, 150~300r/min of rotating speed, 30h is cultivated, obtains level liquid seed culture fluid；
Secondary seed culture：The secondary seed medium prepared in following ratios：Peptone 20g/L, beef extract 5g/L, chlorine Change sodium 5g/L, dipotassium hydrogen phosphate 1g/L, pH 7.0；The level liquid seed culture fluid of gained is seeded to the secondary seed training Support in base, under the conditions of rotating speed 200r/min, throughput 0.8vvm, 28 DEG C of temperature, cultivate 32h, obtain the training of secondary liquid seed Nutrient solution；
Fermented and cultured：Liquid fermentation medium is prepared in following ratios：Peptone 20g/L, beef extract 5g/L, yeast extract 5g/L, potassium dihydrogen phosphate 1g/L, magnesium sulfate 0.5g/L, pH 7.4；With the inoculum concentration of liquid fermentation medium weight 8%, by institute The secondary liquid seed culture fluid obtained is inoculated in the liquid fermentation medium, in speed of agitator 150r/min, throughput 0.7vvm, under the conditions of 38 DEG C of fermentation temperature, fermented and cultured 36h, obtain fermentation of bacillus subtilis nutrient solution；
It is prepared by bacterium powder：The fermentation of bacillus subtilis medium centrifugal of gained is separated and collects wet bacillus subtilis Body, after mannitol protective agent, Tween 80 are mixed with wet bacillus subtilis thalline, add macropore starch carrier and adsorbed, then Crushed after low temperature drying and bacillus subtilis powder is made；Wherein, the weight of the mannitol protective agent and wet bacillus subtilis thalline Amount is than being 0.2:1(g/Kg)；The weight ratio of the Tween 80 and wet bacillus subtilis thalline is 10:1(g/Kg)；The macropore forms sediment The weight ratio of powder carrier and wet bacillus subtilis thalline is 0.5:1(Kg/Kg).
(2) bacillus coagulans is to be made using following methods：
Inclined-plane culture：The slant medium prepared in following ratios：Peptone 10g/L, beef extract 3g/L, sodium chloride 5g/ L, agar 18g/L, pH 7.2；Bacillus coagulans kind is seeded on slant medium, under the conditions of 32 DEG C of temperature, culture 36h, obtain bacillus coagulans kind；
First order seed culture：The primary-seed medium prepared in following ratios：Peptone 10g/L, beef extract 2g/L, chlorine Change sodium 4g/L, pH 7.2；The bacillus coagulans kind of gained is seeded in the primary-seed medium, 32 DEG C of temperature, Under the conditions of rotating speed 200r/min, 25h is cultivated, obtains level liquid seed culture fluid；
Secondary seed culture：The secondary seed medium prepared in following ratios：Peptone 10g/L, beef extract 3g/L, chlorine Change sodium 5g/L, pH 7.4；The level liquid seed culture fluid of gained is seeded in the secondary seed medium, in rotating speed Under the conditions of 200r/min, 32 DEG C of temperature, 24h is cultivated, obtains secondary liquid seed culture fluid；
Fermented and cultured：Liquid fermentation medium is prepared in following ratios：Glucose 25g/L, peptone 15g/L, dusty yeast 10g/L, potassium dihydrogen phosphate 1g/L, magnesium sulfate 0.5g/L, pH 7.2；With the inoculum concentration of liquid fermentation medium weight 8%, by two Level liquid seeds nutrient solution is inoculated in the liquid fermentation medium, in 40 DEG C of speed of agitator 150r/min, fermentation temperature bars Under part, fermented and cultured 24h, bacillus coagulans fermentation culture is obtained；
It is prepared by bacterium powder：Bacillus coagulans fermentation culture is centrifuged and collects wet bacillus coagulans body, will be sweet After dew alcohol protective agent, Tween 80 mix with wet bacillus coagulans body, add macropore starch carrier and adsorbed, then low temperature drying Crush afterwards and bacillus coagulans powder is made；Wherein, the weight ratio of the mannitol protective agent and wet bacillus coagulans body is 0.3:1(g/Kg)；The weight ratio of the Tween 80 and wet bacillus coagulans body is 15:1(g/Kg)；The macropore starch carrier Weight ratio with wet bacillus coagulans body is 0.6:1(Kg/Kg).
(3) saccharomycete is to be made using following methods：
Inclined-plane culture：By barms be inoculated in sugared content 10%, pH 6.0 brewer's wort slant medium on, at 28 DEG C Under the conditions of, 40h is cultivated, obtains inclined-plane barms；
Level liquid seed culture：Sugared content 15%, pH 7.0 brewer's wort are inoculated in after inclined-plane barms is diluted In culture medium, under the conditions of 30 DEG C, 36h is cultivated, obtains level liquid seed culture fluid；
Secondary liquid seed culture：Level liquid seed culture fluid is inoculated in sugared content 20%, pH7.0 brewer's wort is trained Support in base, under the conditions of 36 DEG C, cultivate 48h, obtain secondary liquid seed culture fluid；
Fermented and cultured：Liquid fermentation medium is prepared in following ratios：Glucose 20g/L, peptone 10g/L, di(2-ethylhexyl)phosphate Hydrogen potassium 3g/L, magnesium sulfate 2g/L, ferrous sulfate 5g/L, manganese sulfate 4g/L；With the inoculum concentration of liquid fermentation medium weight 8%, Secondary liquid seed culture fluid is inoculated in liquid fermentation medium, 30 DEG C of cultivation temperature, pH 6.2, pressure 0.05MPa, Under conditions of rotating speed 300r/min, 48h is cultivated, obtains saccharomycetes to make fermentation nutrient solution；
It is prepared by bacterium powder：Saccharomycetes to make fermentation medium centrifugal is separated and collects wet yeast thalline, by mannitol protective agent, is told After temperature 80 mixes with wet yeast thalline, add macropore starch carrier and adsorbed, then crushed after low temperature drying and yeast bacterium powder is made； Wherein, the weight ratio of the mannitol protective agent and wet yeast thalline is 0.2:1(g/Kg)；The Tween 80 and wet yeast thalline Weight ratio be 12:1(g/Kg)；The weight ratio of the macropore starch carrier and wet yeast thalline is 0.5:1(Kg/Kg).
(4) raw material of following percentage by weight meters is taken：Imported fish meal 45%, fermented bean dregs 42%, small peptide 0.5%, withered grass Bacillus 5%, bacillus coagulans 2%, saccharomycete 2%, digestive ferment 0.5%, organic acid 2%, (its of somatomedin 1% In, the digestive ferment is cellulase, phytase, protease according to 2:2:1 weight ratio mixes；The somatomedin It is vitamin E, copper sulphate, ferrous sulfate, manganese sulfate according to 1:1:1:1 weight ratio is mixed)；By obtained by step (1) The saccharomycete obtained by bacillus coagulans, step (3) obtained by bacillus subtilis, step (2) is mixed into composite bacteria agent, then will The composite bacteria agent of gained, imported fish meal, fermented bean dregs, small peptide, digestive ferment, malic acid, somatomedin mixing, in temperature 25 DEG C, under the conditions of rotating speed 150r/min, 10 days obtained aquatic products high-efficiency active biological fermentation baits of solid fermentation culture.
Embodiment 2
Step (1) in the embodiment of the present invention 1 is repeated, bacillus subtilis is made；Repeat step in the embodiment of the present invention 1 (2) bacillus coagulans, is made；Step (3) in embodiment 1 is repeated, saccharomycete is made.
Take the raw material of following percentage by weight meters：Imported fish meal 42%, fermented bean dregs 45%, small peptide 1%, bacillus subtilis Bacterium 2%, bacillus coagulans 3%, saccharomycete 3%, digestive ferment 1%, organic acid 1.5%, (wherein, the institute of somatomedin 1.5% Digestive ferment is stated as cellulase, phytase, protease is according to 2:3:1 weight ratio mixes；The somatomedin is dimension Raw plain E, copper sulphate, ferrous sulfate, manganese sulfate are according to 1:1:1:1 weight ratio is mixed)；By the bacillus subtilis, Bacillus coagulans, saccharomycete are mixed into composite bacteria agent, then by the composite bacteria agent of gained, imported fish meal, fermented bean dregs, small peptide, Digestive ferment, malic acid, somatomedin mixing, under the conditions of 30 DEG C of temperature, rotating speed 200r/min, solid fermentation culture is made for 5 days Obtain aquatic products high-efficiency active biological fermentation bait.
Embodiment 3
Step (1) in the embodiment of the present invention 1 is repeated, bacillus subtilis is made；Repeat step in the embodiment of the present invention 1 (2) bacillus coagulans, is made；Step (3) in embodiment 1 is repeated, saccharomycete is made.
Take the raw material of following percentage by weight meters：Imported fish meal 45%, fermented bean dregs 36%, small peptide 1.5%, withered grass gemma Bacillus 3%, bacillus coagulans 5%, saccharomycete 5%, digestive ferment 0.5%, organic acid 2%, (wherein, the institute of somatomedin 2% Digestive ferment is stated as cellulase, phytase, protease is according to 3:2:1 weight ratio mixes；The somatomedin is dimension Raw plain E, copper sulphate, ferrous sulfate, manganese sulfate are according to 1:1:1:1 weight ratio is mixed)；By the bacillus subtilis, Bacillus coagulans, saccharomycete are mixed into composite bacteria agent, then by the composite bacteria agent of gained, imported fish meal, fermented bean dregs, small peptide, Digestive ferment, malic acid, somatomedin mixing, under the conditions of 35 DEG C of temperature, rotating speed 400r/min, solid fermentation culture is made for 3 days Obtain aquatic products high-efficiency active biological fermentation bait.
Embodiment 4
Step (1) in the embodiment of the present invention 1 is repeated, bacillus subtilis is made；Repeat step in the embodiment of the present invention 1 (2) bacillus coagulans, is made.
Take the raw material of following percentage by weight meters：Imported fish meal 30%, fermented bean dregs 30%, enzymolysis protein 30%, small peptide 0.5%th, bacillus subtilis 1%, bacillus coagulans 6%, digestive ferment 0.5%, organic acid 1%, (its of somatomedin 1% In, the digestive ferment is cellulase, phytase, protease according to 3:2:1 weight ratio mixes；The somatomedin It is vitamin E, copper sulphate, ferrous sulfate, manganese sulfate according to 1:1:1:1 weight ratio is mixed)；By the bacillus subtilis Bacterium, bacillus coagulans are mixed into composite bacteria agent, then by the composite bacteria agent of gained, imported fish meal, fermented bean dregs, enzymolysis protein, Small peptide, digestive ferment, malic acid, somatomedin mixing, under the conditions of 25 DEG C of temperature, rotating speed 200r/min, solid fermentation culture 4 Its obtained aquatic products high-efficiency active biological fermentation bait.
Embodiment 5
Step (1) in the embodiment of the present invention 1 is repeated, bacillus subtilis is made；Repeat step in the embodiment of the present invention 1 (2) bacillus coagulans, is made.
Take the raw material of following percentage by weight meters：Imported fish meal 30%, fermented bean dregs 40%, enzymolysis protein 20%, small peptide 1%th, bacillus subtilis 2%, bacillus coagulans 4%, digestive ferment 0.5%, organic acid 1%, (its of somatomedin 1.5% In, the digestive ferment is cellulase, phytase, protease according to 4:3:1 weight ratio mixes；The somatomedin It is vitamin E, copper sulphate, ferrous sulfate, manganese sulfate according to 1:1:1:1 weight ratio is mixed)；By the bacillus subtilis Bacterium, bacillus coagulans are mixed into composite bacteria agent, then by the composite bacteria agent of gained, imported fish meal, fermented bean dregs, enzymolysis protein, Small peptide, digestive ferment, malic acid, somatomedin mixing, under the conditions of 28 DEG C of temperature, rotating speed 250r/min, solid fermentation culture 5 Its obtained aquatic products high-efficiency active biological fermentation bait.
Embodiment 6
Step (1) in the embodiment of the present invention 1 is repeated, bacillus subtilis is made；Repeat step in the embodiment of the present invention 1 (2) bacillus coagulans, is made.
Take the raw material of following percentage by weight meters：Imported fish meal 35%, fermented bean dregs 45%, enzymolysis protein 12%, small peptide 1%th, bacillus subtilis 3%, bacillus coagulans 2.5%, digestive ferment 0.5%, organic acid 0.5%, somatomedin 0.5% (wherein, the digestive ferment is cellulase, phytase, protease according to 4:2:1 weight ratio mixes；The growth promotion The factor is vitamin E, copper sulphate, ferrous sulfate, manganese sulfate according to 1:1:1:1 weight ratio is mixed)；By the withered grass bud Spore bacillus, bacillus coagulans are mixed into composite bacteria agent, then by the composite bacteria agent of gained, imported fish meal, fermented bean dregs, enzymolysis egg In vain, small peptide, digestive ferment, malic acid, somatomedin mixing, under the conditions of 35 DEG C of temperature, rotating speed 400r/min, solid fermentation training Support 3 days obtained aquatic products high-efficiency active biological fermentation baits.
Embodiment 7
Step (1) in the embodiment of the present invention 1 is repeated, bacillus subtilis is made；Repeat step in the embodiment of the present invention 1 (2) bacillus coagulans, is made；Step (3) in embodiment 1 is repeated, saccharomycete is made.
Take the raw material of following percentage by weight meters：Imported fish meal 40%, fermented bean dregs 32%, enzymolysis protein 10%, small peptide 1.5%th, bacillus subtilis 5%, bacillus coagulans 8%, saccharomycete 1%, digestive ferment 0.5%, organic acid 1%, growth promotion (wherein, the digestive ferment is cellulase, phytase, protease according to 2 to the factor 1%:4:1 weight ratio mixes；It is described Somatomedin is vitamin E, copper sulphate, ferrous sulfate, manganese sulfate according to 1:1:1:1 weight ratio is mixed)；By described in Bacillus subtilis, bacillus coagulans, saccharomycete are mixed into composite bacteria agent, then composite bacteria agent, imported fish meal, hair by gained Ferment dregs of beans, enzymolysis protein, small peptide, digestive ferment, malic acid, somatomedin mixing, in 27 DEG C of temperature, rotating speed 250r/min conditions Under, 7 days obtained aquatic products high-efficiency active biological fermentation baits of solid fermentation culture.
Embodiment 8
Step (1) in the embodiment of the present invention 1 is repeated, bacillus subtilis is made；Repeat step in the embodiment of the present invention 1 (2) bacillus coagulans, is made；Step (3) in embodiment 1 is repeated, saccharomycete is made.
Take the raw material of following percentage by weight meters：Imported fish meal 40%, fermented bean dregs 25%, enzymolysis protein 15%, small peptide 0.5%th, bacillus subtilis 5%, bacillus coagulans 6%, saccharomycete 4%, digestive ferment 0.5%, organic acid 2%, growth promotion (wherein, the digestive ferment is cellulase, phytase, protease according to 5 to the factor 2%:4:1 weight ratio mixes；It is described Somatomedin is vitamin E, copper sulphate, ferrous sulfate, manganese sulfate according to 1:1:1:1 weight ratio is mixed)；By described in Bacillus subtilis, bacillus coagulans, saccharomycete are mixed into composite bacteria agent, then composite bacteria agent, imported fish meal, hair by gained Ferment dregs of beans, enzymolysis protein, small peptide, digestive ferment, malic acid, somatomedin mixing, in 25 DEG C of temperature, rotating speed 400r/min conditions Under, 5 days obtained aquatic products high-efficiency active biological fermentation baits of solid fermentation culture.
Comparative example 1
Step (1) in the embodiment of the present invention 1 is repeated, bacillus subtilis is made.
Take the raw material of following percentage by weight meters：Imported fish meal 30%, fermented bean dregs 40%, enzymolysis protein 20%, small peptide 1%th, (wherein, the digestive ferment is fibre for bacillus subtilis 6%, digestive ferment 0.5%, organic acid 1%, somatomedin 1.5% Plain enzyme, phytase, protease are tieed up according to 4:3:1 weight ratio mixes；The somatomedin be vitamin E, copper sulphate, Ferrous sulfate, manganese sulfate are according to 1:1:1:1 weight ratio is mixed)；By the bacillus subtilis, imported fish meal, fermentation Dregs of beans, enzymolysis protein, small peptide, digestive ferment, malic acid, somatomedin mixing, in 28 DEG C of temperature, rotating speed 250r/min conditions Under, 5 days obtained aquatic products high-efficiency active biological fermentation baits of solid fermentation culture.
Comparative example 2
Step (1) in the embodiment of the present invention 1 is repeated, bacillus subtilis is made；Repeat step in the embodiment of the present invention 1 (3) saccharomycete, is made.
Take the raw material of following percentage by weight meters：Imported fish meal 30%, fermented bean dregs 40%, enzymolysis protein 20%, small peptide 1%th, bacillus subtilis 2%, saccharomycete 4%, digestive ferment 0.5%, organic acid 1%, somatomedin 1.5% are (wherein, described Digestive ferment is cellulase, phytase, protease according to 4:3:1 weight ratio mixes；The somatomedin is given birth to for dimension Plain E, copper sulphate, ferrous sulfate, manganese sulfate are according to 1:1:1:1 weight ratio is mixed)；By the bacillus subtilis, ferment Female bacterium is mixed into composite bacteria agent, then by the composite bacteria agent of gained, imported fish meal, fermented bean dregs, enzymolysis protein, small peptide, digestive ferment, Malic acid, somatomedin mixing, under the conditions of 28 DEG C of temperature, rotating speed 250r/min, 5 days obtained aquatic products of solid fermentation culture are used High-efficiency active biological fermentation bait.
Comparative example 3
Step (1) in the embodiment of the present invention 1 is repeated, bacillus subtilis is made；Repeat step in the embodiment of the present invention 1 (2) bacillus coagulans, is made.
Take the raw material of following percentage by weight meters：Imported fish meal 30%, fermented bean dregs 40%, enzymolysis protein 20%, small peptide 1%th, bacillus subtilis 2%, bacillus coagulans 4%, digestive ferment 0.5%, organic acid 1%, (its of somatomedin 1.5% In, the digestive ferment is cellulase, phytase, protease according to 4:3:1 weight ratio mixes；The somatomedin It is vitamin E, copper sulphate, ferrous sulfate, manganese sulfate according to 1:1:1:1 weight ratio is mixed)；By the bacillus subtilis Bacterium, bacillus coagulans, saccharomycete are mixed into composite bacteria agent, then by the composite bacteria agent of gained, imported fish meal, fermented bean dregs, enzyme Solve albumen, small peptide, digestive ferment, malic acid, somatomedin mixing, under the conditions of 28 DEG C of temperature, rotating speed 250r/min, solid hair 5 days obtained aquatic products high-efficiency active biological fermentation baits of ferment culture.
Illustrate aquatic products of the invention with high-efficiency active biological fermentation bait in cultivation fish, shrimp with reference to specific data Effect：
Effect example 1 feeds Penaeus Vannmei experiment
In April, 2016, the biological aquatic farm of intelligence dragon in Guangzhou, from 13 mouthfuls of ponds, every mouthful of pond water surface area is about For 2 mu, it is divided into 13 groups, wherein, 12 groups are test group, and one group is control group；According to the throw-in density of 50,000 tails/mu, will be about The Penaeus Vannmei shrimp seedling of 3.0 centimetres of body long sizes is launched in 12 mouthfuls of pond waters, and test group and control group are according to routine Method cultivates Penaeus Vannmei, unlike：In test group, with 0.25 times of commercially available honest shrimp feed weight, the present invention is carried For aquatic products high-efficiency active biological fermentation bait be uniformly mixed with commercially available honest shrimp feed after feed again to test group In pond waters；Control group only feeds Penaeus Vannmei using commercially available honest shrimp feed, and specific packet is as shown in table 2；Experiment (90 days) period observed the health condition of Penaeus Vannmei, collected dead shrimp from blowdown pipe sleeve net daily, and it is right in vain to count South America Survival rate, the output value of shrimp, statistical result is as shown in Fig. 1~2.
The experiment packet of the culture of Penaeus vannamei of table 1
Packet Feed feed
Test group 1 The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 1
Test group 2 The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 2
Test group 3 The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 3
Test group 4 The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 4
Test group 5 The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 5
Test group 6 The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 6
Test group 7 The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 7
Test group 8 The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 8
Test group 9 The product of the gained of commercially available honest shrimp feed+comparative example of the present invention 1
Test group 10 The product of the gained of commercially available honest shrimp feed+comparative example of the present invention 2
Test group 11 The product of the gained of commercially available honest shrimp feed+comparative example of the present invention 3
Control group Commercially available honest shrimp feed
From figure 3, it can be seen that in test group 1~8 of the present invention the survival rate of Penaeus Vannmei more than 76%, wherein, The survival rate highest of the Penaeus Vannmei of test group 5, up to 84.2%, and Penaeus Vannmei is not higher than in test group 9~11 66.5%, Penaeus Vannmei is only 58.3% in control group, and the aquatic products that this explanation embodiment of the present invention 1~8 is provided is with efficient Active bio fermentation bait is advantageous to improve the immunity of Penaeus Vannmei.
From fig. 4, it can be seen that in test group 1~8 of the present invention the output value of Penaeus Vannmei more than 500Kg/ mus, its In, the output value highest of the Penaeus Vannmei of test group 5, reach more than 615Kg/ mus, and Penaeus Vannmei is equal in test group 9~11 Less than 400Kg/ mus, Penaeus Vannmei is less than 330Kg/ mus in control group, the water that this explanation embodiment of the present invention 6~14 is provided Production high-efficiency active biological fermentation bait is advantageous to improve the function of intestinal canal of Penaeus Vannmei, improves food conversion ratio, and improves life The intestinal microecology balance of Penaeus Vannmei.
Total described, the aquatic products that the embodiment of the present invention 1~8 is provided is that South America is right in vain with high-efficiency active biological fermentation bait The formation of the intestinal environment of shrimp creates good condition, by aerobic bacteria producing bacillus subtilis give birth to more nutriments and Digestive ferment (amylase, protease etc.), while the oxygen in Penaeus Vannmei enteron aisle can also be exhausted, it is bacillus coagulans, ferment The breeding of female bacterium creates good growing environment, is advantageous to improve the immunity of Penaeus Vannmei；It is white to be advantageous to improvement South America The function of intestinal canal of prawn, improves food conversion ratio, and improves the intestinal microecology balance of Penaeus Vannmei.
Effect example 2 feeds Rofe fish test
In April, 2016, the biological aquatic farm of intelligence dragon in Guangzhou, from 12 mouthfuls of ponds, every mouthful of pond water surface area is about For 2 mu, it is divided into 12 groups, wherein, 11 groups are test group, and one group is control group；According to the throw-in density of 8000 tails/mu, by 100 The Tilapia mossambica of tail/Kg specifications is launched in 12 mouthfuls of pond waters, test group and control group conventionally cultivating tilapia, Unlike：In test group, with 0.1 times of commercially available honest fish meal weight, by aquatic products provided by the invention with high-efficiency activated life Thing fermentation bait is fed into test group pond waters again after being uniformly mixed with commercially available honest fish meal；Control group only makes Tilapia mossambica is fed with commercially available honest fish meal, specific packet is as shown in table 2；The health of (90 days) observation Tilapia mossambicas during experiment Situation, dead fish is collected from blowdown pipe sleeve net daily, and count the survival rate of Tilapia mossambica, the output value, the institute of statistical result such as Fig. 3~4 Show.
The Tilapia mossambica culture experiment of table 2 is grouped
Packet Feed feed
Test group 1 The product of the commercially available gained of honest fish meal+embodiment of the present invention 1
Test group 2 The product of the commercially available gained of honest fish meal+embodiment of the present invention 2
Test group 3 The product of the commercially available gained of honest fish meal+embodiment of the present invention 3
Test group 4 The product of the commercially available gained of honest fish meal+embodiment of the present invention 4
Test group 5 The product of the commercially available gained of honest fish meal+embodiment of the present invention 5
Test group 6 The product of the commercially available gained of honest fish meal+embodiment of the present invention 6
Test group 7 The product of the commercially available gained of honest fish meal+embodiment of the present invention 7
Test group 8 The product of the commercially available gained of honest fish meal+embodiment of the present invention 8
Test group 9 The product of the gained of commercially available honest fish meal+comparative example of the present invention 1
Test group 10 The product of the gained of commercially available honest fish meal+comparative example of the present invention 2
Test group 11 The product of the gained of commercially available honest fish meal+comparative example of the present invention 3
Control group Commercially available honest fish meal
It will be seen from figure 1 that in test group 1~8 of the present invention the survival rate of Tilapia mossambica more than 85%, wherein, experiment The survival rate highest of the Tilapia mossambica of group 2, reaches more than 93%, and Tilapia mossambica is below 80% in test group 9~11, in control group Tilapia mossambica is only 63.1%, and the aquatic products that this explanation embodiment of the present invention 1~8 is provided is favourable with high-efficiency active biological fermentation bait In the immunity for improving Tilapia mossambica.
Figure it is seen that in test group 1~8 of the present invention the output value of Tilapia mossambica more than 4100Kg/ mus, wherein, examination The output value highest of the Tilapia mossambica of group 2 is tested, is reached more than 4750Kg/ mus, and Tilapia mossambica is below 3800Kg/ in test group 9~11 Mu, Tilapia mossambica is less than 2500Kg/ mus in control group, the high-efficiency activated life of the aquatic products that this explanation embodiment of the present invention 1~8 is provided Thing fermentation bait is advantageous to improve the function of intestinal canal of Tilapia mossambica, improves food conversion ratio, the intestinal microecology for improving Tilapia mossambica is put down Weighing apparatus.
Total described, the aquatic products that the embodiment of the present invention 1~8 is provided is Tilapia mossambica with high-efficiency active biological fermentation bait The formation of intestinal environment creates good condition, and more nutriments and digestion are given birth to by aerobic bacteria producing bacillus subtilis Enzyme (amylase, protease etc.), while the oxygen in Tilapia mossambica enteron aisle can also be exhausted, it is the breeding of bacillus coagulans, yeast Good growing environment is created, is advantageous to improve the immunity of Tilapia mossambica；Be advantageous to improve the function of intestinal canal of Tilapia mossambica, improve Feed conversion rate, improve the intestinal microecology balance of Tilapia mossambica.
Embodiments of the present invention are explained in detail above in association with accompanying drawing, but the invention is not restricted to described implementation Mode.For a person skilled in the art, in the case where not departing from the principle of the invention and spirit, to these embodiments A variety of change, modification, replacement and modification are carried out, are still fallen within protection scope of the present invention.

Claims (10)

  1. A kind of 1. aquatic products high-efficiency active biological fermentation bait, it is characterised in that：Aquatic products high-efficiency active biological fermentation bait The raw material of material, by weight percentage, including：Imported fish meal 30~45%, fermented bean dregs 25~45%, enzymolysis protein 0~ 30%th, small peptide 0.5~1.5%, bacillus subtilis 1~5%, bacillus coagulans 2~8%, saccharomycete 0~5%, digestive ferment 0.1~1%, organic acid 0.5~2%, somatomedin 0.5~2%.
  2. 2. aquatic products according to claim 1 high-efficiency active biological fermentation bait, it is characterised in that：The aquatic products is with efficiently Active bio fermentation bait active bacteria number is not less than 1,000,000,000/gram.
  3. 3. aquatic products according to claim 1 high-efficiency active biological fermentation bait, it is characterised in that：The digestive ferment is selected from One or more in cellulase, amylase, phytase.
  4. 4. aquatic products according to claim 1 high-efficiency active biological fermentation bait, it is characterised in that：The organic acid is selected from One or more in fumaric acid, lactic acid, citric acid, malic acid, tartaric acid, sorbic acid, acetic acid, formic acid, propionic acid.
  5. 5. aquatic products according to claim 1 high-efficiency active biological fermentation bait, it is characterised in that：The somatomedin One or more in zinc sulfate, sodium selenite, vitamin E, calcium iodate, copper sulphate, ferrous sulfate, manganese sulfate.
  6. 6. aquatic products according to claim 1 high-efficiency active biological fermentation bait, it is characterised in that：The bacillus subtilis Bacterium is to be made using following methods：
    (1) inclined-plane culture：The slant medium prepared in following ratios：15~30g/L of glucose, 10~20g/L of peptone, chlorine Change 0.5~2g/L of sodium, 3~10g/L of yeast extract, agar 15~30g/L, pH 6.5~7.0；Bacillus subtilis strain is seeded to On slant medium, under the conditions of 25~37 DEG C of temperature, 24~48h is cultivated, obtains bacillus subtilis strain；
    (2) first order seed culture：The primary-seed medium prepared in following ratios：10~30g/L of glucose, peptone 10~ 20g/L, 0.5~2g/L of sodium chloride, yeast extract 3~10g/L, pH 6.5~7.0；By the bacillus subtilis obtained by step (1) Kind is seeded in the primary-seed medium, under the conditions of 25~37 DEG C of temperature, 150~300r/min of rotating speed, culture 24~ 36h, obtain level liquid seed culture fluid；
    (3) secondary seed culture：The secondary seed medium prepared in following ratios：10~30g/L of peptone, beef extract 3~ 10g/L, 3~7g/L of sodium chloride, dipotassium hydrogen phosphate 0.5~2g/L, pH 6.5~7.0；By the level liquid kind obtained by step (2) Sub- nutrient solution is seeded in the secondary seed medium, in 180~250r/min of rotating speed, 0.5~1.0vvm of throughput, temperature Under the conditions of 25~37 DEG C, 24~36h is cultivated, obtains secondary liquid seed culture fluid；
    (4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：10~30g/L of peptone, 3~10g/L of beef extract, 3~10g/L of yeast extract, 0.5~2g/L of potassium dihydrogen phosphate, magnesium sulfate 0.1~1g/L, pH 7.0~7.5；With liquid fermentation and culture The inoculum concentration of base weight 6~10%, the secondary liquid seed culture fluid obtained by step (3) is inoculated in the liquid fermentation and culture In base, under the conditions of 100~200r/min of speed of agitator, 0.5~0.8vvm of throughput, 35~40 DEG C of fermentation temperature, fermentation training 24~48h is supported, obtains fermentation of bacillus subtilis nutrient solution；
    (5) prepared by bacterium powder：Fermentation of bacillus subtilis medium centrifugal obtained by step (4) is separated and collects wet withered grass gemma Bacillus body, after mannitol protective agent, Tween 80 are mixed with wet bacillus subtilis thalline, add macropore starch carrier and inhaled It is attached, then crushed after low temperature drying and bacillus subtilis powder is made；Wherein, the mannitol protective agent and wet bacillus subtilis thalline Weight ratio be 0.15~0.5:1(g/Kg)；The weight ratio of the Tween 80 and wet bacillus subtilis thalline is 5~20:1(g/ Kg)；The weight ratio of the macropore starch carrier and wet bacillus subtilis thalline is 0.2~1:1(Kg/Kg).
  7. 7. aquatic products according to claim 1 high-efficiency active biological fermentation bait, it is characterised in that：The condensation gemma bar Bacterium is to be made using following methods：
    (1) inclined-plane culture：The slant medium prepared in following ratios：5~15g/L of peptone, 1~5g/L of beef extract, chlorination 2~7g/L of sodium, agar 15~20g/L, pH 7.2~7.4；Bacillus coagulans kind is seeded on slant medium, in temperature Under the conditions of 25~37 DEG C, 24~48h is cultivated, obtains bacillus coagulans kind；
    (2) first order seed culture：The primary-seed medium prepared in following ratios：5~15g/L of peptone, beef extract 1~ 5g/L, 2~7g/L of sodium chloride, pH7.2~7.4；Bacillus coagulans kind obtained by step (1) is seeded to the first order seed In culture medium, under the conditions of 25~37 DEG C of temperature, 150~300r/min of rotating speed, 18~30h is cultivated, obtains level liquid seed Nutrient solution；
    (3) secondary seed culture：The secondary seed medium prepared in following ratios：5~15g/L of peptone, beef extract 1~ 5g/L, 2~7g/L of sodium chloride, pH7.2~7.4；Level liquid seed culture fluid obtained by step (2) is seeded to the two level In seed culture medium, under the conditions of 180~250r/min of rotating speed, 0.5~1.0vvm of throughput, 25~37 DEG C of temperature, culture 18 ~30h, obtain secondary liquid seed culture fluid；
    (4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：20~30g/L of glucose, 10~20g/L of peptone, 5~15g/L of dusty yeast, 0.5~2g/L of potassium dihydrogen phosphate, magnesium sulfate 0.1~1g/L, pH 7.0~7.5；With liquid fermentation and culture The inoculum concentration of base weight 6~10%, the secondary liquid seed culture fluid obtained by step (3) is inoculated in the liquid fermentation and culture In base, under the conditions of 100~200r/min of speed of agitator, 0.5~0.8vvm of throughput, 35~50 DEG C of fermentation temperature, fermentation training 20~30h is supported, obtains bacillus coagulans fermentation culture；
    (5) prepared by bacterium powder：Bacillus coagulans fermentation culture obtained by step (4) is centrifuged and collects wet condensation gemma Bacillus body, after mannitol protective agent, Tween 80 are mixed with wet bacillus coagulans body, add macropore starch carrier and inhaled It is attached, then crushed after low temperature drying and bacillus coagulans powder is made；Wherein, the mannitol protective agent and wet bacillus coagulans body Weight ratio be 0.15~0.5:1(g/Kg)；The weight ratio of the Tween 80 and wet bacillus coagulans body is 5~20:1(g/ Kg)；The weight ratio of the macropore starch carrier and wet bacillus coagulans body is 0.2~1:1(Kg/Kg).
  8. 8. aquatic products according to claim 1 high-efficiency active biological fermentation bait, it is characterised in that：The saccharomycete is to adopt It is made with following methods：
    (1) inclined-plane culture：Barms is inoculated in the brewer's wort slant medium of sugared content 5~20%, pH value 4.0~7.0 On, under the conditions of 25~36 DEG C, 25~50h is cultivated, obtains inclined-plane barms；
    (2) level liquid seed culture：To be inoculated in after the dilution of inclined-plane barms obtained by step (1) sugared content 5~20%, In pH 4.0~7.0 malt extract medium, under the conditions of 25~36 DEG C, 25~50h is cultivated, obtains level liquid seed culture Liquid；
    (3) secondary liquid seed culture：By the level liquid seed culture fluid obtained by step (2) be inoculated in sugared content 5~20%, In pH 4.0~7.0 malt extract medium, under the conditions of 25~36 DEG C, 25~50h is cultivated, obtains secondary liquid seed culture Liquid；
    (4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：10~30g/L of glucose, 5~15g/L of peptone, 1~5g/L of potassium dihydrogen phosphate, 0.1~0.5g/L of magnesium sulfate, 0.3~0.6g/L of ferrous sulfate, 0.3~0.6g/L of manganese sulfate；With The inoculum concentration of liquid fermentation medium weight 6~10%, the secondary liquid seed culture fluid obtained by step (3) is inoculated in liquid In fermentation medium, in 28~32 DEG C of cultivation temperature, pH 5.5~6.5,0.03~0.06MPa of pressure, 200~350r/ of rotating speed Under the conditions of min, 25~50h of fermented and cultured, saccharomycetes to make fermentation nutrient solution is obtained；
    (5) prepared by bacterium powder：Saccharomycetes to make fermentation medium centrifugal obtained by step (4) is separated and collects wet yeast thalline, will be sweet After dew alcohol protective agent, Tween 80 mix with wet yeast thalline, add macropore starch carrier and adsorbed, then crushed after low temperature drying Yeast bacterium powder is made；Wherein, the weight ratio of the mannitol protective agent and wet yeast thalline is 0.15~0.5:1(g/Kg)；Institute The weight ratio for stating Tween 80 and wet yeast thalline is 5~20:1(g/Kg)；The macropore starch carrier and the weight of wet yeast thalline Amount is than being 0.2~1:1(Kg/Kg).
  9. 9. a kind of method of the aquatic products prepared described in any one of claim 1~8 high-efficiency active biological fermentation bait, its feature It is：Comprise the following steps：
    Bacillus subtilis, bacillus coagulans, saccharomycete are mixed into composite bacteria agent, then by the composite bacteria agent of gained, import Fish meal, fermented bean dregs, enzymolysis protein, small peptide, digestive ferment, organic acid, somatomedin mixing, fermented obtained aquatic products is with efficiently Active bio fermentation bait.
  10. 10. a kind of aquatic products high-efficiency active biological fermentation bait as described in any one of claim 1~8 is in feed is prepared Using.
CN201711092012.1A 2017-11-08 2017-11-08 A kind of aquatic products high-efficiency active biological fermentation bait and its preparation method and application Pending CN107712462A (en)

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CN110839757A (en) * 2019-12-04 2020-02-28 北京市水产技术推广站 Goldfish fry starter feed and preparation method thereof
CN112655845A (en) * 2020-12-25 2021-04-16 江苏苏港和顺生物科技有限公司 Preparation method of fermented fish feed replacing antibiotic feed
CN113854252A (en) * 2021-10-28 2021-12-31 淮阴工学院 Novel protein peptide bait for fishing crucian and preparation method thereof
CN114504065A (en) * 2022-03-09 2022-05-17 湖北丰甜生物科技有限公司 Method for producing natural bait low-temperature composite algae for aquatic animals

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CN101647524A (en) * 2009-08-14 2010-02-17 淮安华威饲料发展有限公司 Feed for controlling medicine residues of channel catfishes
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CN110839757A (en) * 2019-12-04 2020-02-28 北京市水产技术推广站 Goldfish fry starter feed and preparation method thereof
CN112655845A (en) * 2020-12-25 2021-04-16 江苏苏港和顺生物科技有限公司 Preparation method of fermented fish feed replacing antibiotic feed
CN113854252A (en) * 2021-10-28 2021-12-31 淮阴工学院 Novel protein peptide bait for fishing crucian and preparation method thereof
CN114504065A (en) * 2022-03-09 2022-05-17 湖北丰甜生物科技有限公司 Method for producing natural bait low-temperature composite algae for aquatic animals

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