CN107821789A - A kind of biologic ferment for improving fishes and shrimps intestinal health degree and its preparation method and application - Google Patents
A kind of biologic ferment for improving fishes and shrimps intestinal health degree and its preparation method and application Download PDFInfo
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- CN107821789A CN107821789A CN201711092042.2A CN201711092042A CN107821789A CN 107821789 A CN107821789 A CN 107821789A CN 201711092042 A CN201711092042 A CN 201711092042A CN 107821789 A CN107821789 A CN 107821789A
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- shrimps
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- intestinal health
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Classifications
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- A—HUMAN NECESSITIES
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- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- 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
- A23K20/116—Heterocyclic compounds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
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- A23K20/163—Sugars; Polysaccharides
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
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Abstract
The invention discloses a kind of biologic ferment for improving fishes and shrimps intestinal health degree and its preparation method and application.The raw material of the biologic ferment for improving fishes and shrimps intestinal health degree, by weight percentage, including：Bacillus coagulans 15~25%, lactobacillus acidophilus 5~15%, Lactobacillus plantarum 0~10%, bacillus subtilis 5~15%, bacillus licheniformis 0~15%, synergist 1~5%, activator 1~5%, carrier 30 as one kind~55%.The present invention use liquid deep layer fermenting technology, can significantly improve live body bacterium number in product, and always the bacterium number of viable bacteria is not less than 4,000,000,000/gram in product.Five kinds of bacillus coagulans, Lactobacillus plantarum, bacillus subtilis, lactobacillus acidophilus, bacillus licheniformis strains are carried out seed selection by the present invention respectively, and mixed with activator, synergist, carrier with good proportioning, form it into optimal compositely proportional, be advantageous to improve fish, the function of intestinal canal of shrimp, improve food conversion ratio, improve fish, shrimp intestinal microecology balance, greatly improve the survival rate and the output value of fish, shrimp.
Description
Technical field
The present invention relates to technical field of aquaculture feedstuff, more particularly to a kind of biological ferment for improving fishes and shrimps intestinal health degree
Element and its preparation method and application.
Background technology
The digestive system of animal particularly enteral have hundreds of, tens billion of bacteriums formed intestinal flora, it is good for animal
Health has close relationship.Intestinal flora can also suppress pathogen propagation while various nutritional physiology effects are played
Deng the effect of defence infection.The generation of the coliform disease of domestic animal, salmonella disease and clostridium enteritis all with enteral
The state of bacterium has substantial connection.Therefore, the balance of intestinal flora is made to keep normally to safeguarding that animal health is essential.It
Raising and improvement with the indices in aquaculture have substantial connection.
Microorganism formulation is a kind of microbial product for substituting or balancing one or more fungus strains in the animal ecosystem.It is narrow
Said in justice, it is that one kind can excite own beneficial culture propagation to increase, while resists the microbial product of harmful fungus strain growth.It is micro-
Biological agent contains the multiple beneficial microorganism such as substantial amounts of lactic acid bacteria, saccharomycete, can be rapid after entering as feed in livestock and poultry body
Breeding, one side is put into the metabolin of strain and enterogenous endotoxin, it is suppressed that the growth of other harmful floras, on the other hand in place
Normal microbial flora is formd in main body, main vitamin is synthesized for host, there is provided nutrition and the invasion for preventing pathogenic bacteria.
Because the abuse of antibiotic causes problems with more prominent in China：Autogenous infection, the increase of resistance strain,
Fish, shrimp immunity reduce, and the destruction of normal intestinal flora and its residual in fishes and shrimps body and in environment etc., the Ministry of Agriculture forbids making
Antibiotic kind is more and more.The development trend of aquaculture is the complete prohibition of antibiotic from now on, therefore antibiotic replaces
The development demand of product is more and more urgent.Probiotics is safe and non-toxic, and with improve production performance, preventing and treating Animal diseases,
Immunologic function is adjusted, improves breeding environment and the multiple functions such as improve food conversion ratio, is overcome using bacterium caused by antibiotic
Group's imbalance, there is the adverse reaction of drug resistance strain and medicine, can not only prevent and cure diseases, and pollution can be reduced and improved
Environment, in actual production process, popularization and application probiotics, there are good economic benefit and social benefit, be advantageous to
Promote the sound development of aquaculture industry of China.
But single microorganism preparation application report in aquaculture is more, on compound micro-ecological preparation especially not
It is less with effect disquisition of the bacterial strain collaboration proportioning in cultivation.Research shows, single microorganism formulation products in the market
There is act on it is single, quality is unstable, cost is high, to improve fishes and shrimps intestinal health degree effect be not so good as antibiotic the defects of.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of biologic ferment for improving fishes and shrimps intestinal health degree and its preparation
Method and its preparation and application, the biologic ferment of the raising fishes and shrimps intestinal health degree have vdiverse in function, steady quality and cost
Low advantage；The immunity of fish, shrimp can be improved, improves fish, the function of intestinal canal of shrimp, improves food conversion ratio again.
In order to solve the above-mentioned technical problem, the technical scheme is that：
In a first aspect, the invention provides a kind of biologic ferment for improving fishes and shrimps intestinal health degree, the raising fishes and shrimps intestines
The raw material of the biologic ferment of road health degree, by weight percentage, including：Bacillus coagulans 15~25%, lactobacillus acidophilus 5
~15%, Lactobacillus plantarum 0~10%, bacillus subtilis 5~15%, bacillus licheniformis 0~15%, synergist 1~
5%th, activator 1~5%, carrier 30 as one kind~55%.
Preferably, total active bacteria number of the biologic ferment for improving fishes and shrimps intestinal health degree is not less than 4,000,000,000/gram.
Preferably, the one kind of the synergist in fumaric acid, lactic acid, citric acid, malic acid, tartaric acid, sorbic acid
It is or several.
Preferably, the activator is selected from amino acid, chromium picolinate, zinc sulfate, sodium selenite, vitamin E, acid iodide
One or more in calcium, copper sulphate, ferrous sulfate, manganese sulfate.
Preferably, the carrier is selected from plant ash, macropore starch, powder of straw, corn flour, rice bran, zeolite powder, lightweight carbonic acid
One or more in calcium, wheat bran, flour, macroporous absorbent resin.
It is further preferred that the carrier includes plant ash, macroporous absorbent resin, wherein, the plant ash is inhaled with macropore
The weight ratio of attached resin is 0.1~0.5:1.
Preferably, the raw material of the biologic ferment for improving fishes and shrimps intestinal health degree, by weight percentage, including：It is solidifying
Tie bacillus 20%, lactobacillus acidophilus 10%, Lactobacillus plantarum 10%, bacillus subtilis 10%, bacillus licheniformis
10%th, synergist 2%, activator 3%, carrier 35%.
Preferably, the raw material of the biologic ferment for improving fishes and shrimps intestinal health degree, by weight percentage, including：It is solidifying
Tie bacillus 21%, lactobacillus acidophilus 12%, bacillus subtilis 12%, synergist 4%, activator 3%, carrier 48%.
Second aspect, present invention also offers a kind of life of the raising fishes and shrimps intestinal health degree prepared as described in relation to the first aspect
The method of thing ferment, comprises the following steps：
The bacillus coagulans, lactobacillus acidophilus, Lactobacillus plantarum, bacillus subtilis, bacillus licheniformis are mixed
Composite bacteria agent is synthesized, then by after the composite bacteria agent of gained, synergist, activator, carrier mixing, is made and improves fishes and shrimps intestinal health
The biologic ferment of degree.
The third aspect, present invention also offers a kind of biological ferment of raising fishes and shrimps intestinal health degree as described in relation to the first aspect
Application of the element in feed is prepared.
Compared with prior art, the beneficial effects of the present invention are：
(1) present invention using liquid deep layer fermenting technology to bacillus coagulans, Lactobacillus plantarum, bacillus subtilis,
Five kinds of lactobacillus acidophilus, bacillus licheniformis strains carry out seed selection respectively, and by improving fermentation batches and craft condition,
Have the advantages that strain survival rate is high, stability is strong, the present invention can significantly improve live body bacterium number in product, total viable bacteria in product
Bacterium number be not less than 4,000,000,000/gram.
(2) it is of the invention by bacillus coagulans, Lactobacillus plantarum, bacillus subtilis, lactobacillus acidophilus, lichens gemma bar
Five kinds of strains of bacterium carry out organic assembling with activator, synergist, carrier, form it into optimal compositely proportional, give full play to each
The flora advantage of strain, good condition is created for the formation of its fish, shrimp intestinal environment, alimentary canal disease can not only be prevented
Disease, and fish, shrimp growth can be promoted, put on weight, reach the purpose of compound synergic.By aerobic bacteria bacillus subtilis and
Bacillus licheniformis interaction produces more nutriments and digestive ferment (such as amylase, protease), while can also exhaust
Oxygen in fish, shrimp enteron aisle, good growth is created for the breeding of bacillus coagulans, Lactobacillus plantarum, lactobacillus acidophilus
Environment, improve fish, the function of intestinal canal of shrimp, improve food conversion ratio, improve fish, shrimp intestinal microecology balance.The raising of the present invention
The biologic ferment of fishes and shrimps intestinal health degree feeds Penaeus Vannmei and sieve as Penaeus Vannmei and the feed addictive of Tilapia mossambica
Non- fish, survival rate respectively reach 81%, more than 85%, and yield is big, good in economic efficiency.
(3) the inventive method is simple, technical maturity, is easy to grasp control, is easy to promote.
Brief description of the drawings
The statistical results chart for the Penaeus Vannmei survival rate that Fig. 1 is cultivated by effect example 1 of the present invention；
The statistical results chart for the Penaeus Vannmei output value that Fig. 2 is cultivated by effect example 1 of the present invention；
The statistical results chart for the Tilapia mossambica survival rate that Fig. 3 is cultivated by effect example 2 of the present invention；
The statistical results chart for the Rofe value of the catch that Fig. 4 is cultivated 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 biologic ferment for improving fishes and shrimps intestinal health degree, the raising fishes and shrimps intestines
The raw material of the biologic ferment of road health degree, by weight percentage, including：Bacillus coagulans 15~25%, lactobacillus acidophilus 5
~15%, Lactobacillus plantarum 0~10%, bacillus subtilis 5~15%, bacillus licheniformis 0~15%, synergist 1~
5%th, activator 1~5%, carrier 30 as one kind~55%.
In embodiments of the present invention, total active bacteria number of the biologic ferment for improving fishes and shrimps intestinal health degree is not less than
4000000000/gram.
In embodiments of the present invention, the synergist is selected from fumaric acid, lactic acid, citric acid, malic acid, tartaric acid, mountain
One or more in pears acid.
In a preferred embodiment of this invention, the synergist is citric acid.
In embodiments of the present invention, the activator is selected from amino acid, chromium picolinate, zinc sulfate, sodium selenite, dimension
One or more in raw plain E, calcium iodate, copper sulphate, ferrous sulfate, manganese sulfate.
In a preferred embodiment of this invention, the activator is vitamin E.
In embodiments of the present invention, the carrier is selected from plant ash, macropore starch, powder of straw, corn flour, rice bran, boiling
One or more in stone flour, precipitated calcium carbonate, wheat bran, flour, macroporous absorbent resin.
In embodiments of the present invention, the carrier include plant ash, macroporous absorbent resin, wherein, the plant ash with
The weight ratio of macroporous absorbent resin is 0.1~0.5:1.
In embodiments of the present invention, the bacillus coagulans, lactobacillus acidophilus, Lactobacillus plantarum, bacillus subtilis
Bacterium, bacillus licheniformis are that liquid deep layer fermenting is made to dry after fermentation culture and is made.
In embodiments of the present invention, the raw material of the biologic ferment for improving fishes and shrimps intestinal health degree, by weight percentage
Than meter, including：Bacillus coagulans 20%, lactobacillus acidophilus 10%, Lactobacillus plantarum 10%, bacillus subtilis 10%,
Clothing bacillus 10%, synergist 2%, activator 3%, carrier 35%.
In embodiments of the present invention, the raw material of the biologic ferment for improving fishes and shrimps intestinal health degree, by weight percentage
Than meter, including：Bacillus coagulans 21%, lactobacillus acidophilus 12%, bacillus subtilis 12%, synergist 4%, activator
3%th, carrier 48%.
In a preferred embodiment of this 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 meter 6~10%, the secondary liquid seed culture fluid obtained by step (3) is inoculated with into the liquid
In fermentation 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 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 lactic acid bacteria, also known as " having spore lactic acid bacteria ".It is through the U.S.
FDA approval one kind " generally believing safety " bacillus class lactic acid bacteria, bacillus coagulans fish, shrimp enteron aisle in, can ferment
Produce a large amount of antibacterial coaguins, lactic acid, amino acid, vitamin and a variety of digestive ferments.The functional characteristics of bacillus coagulans is as follows：
(1) effectively suppress the growth of harmful bacteria, improve intestine microenvironment, promote intestinal growth, strengthen function of intestinal canal；(2) improve
Feed quality, promote digesting and assimilating for feed, reduce feedstuff-meat ratio；(3) be effectively improved weanling pig due to stress and allergic reaction
Caused dysbiosis diarrhea；(4) it is without any side effects, it is safe to use, and the ammonia in environment, hydrogen sulfide etc. can be reduced
Pernicious gas, improve breeding environment.
In a preferred embodiment of this invention, the lactobacillus acidophilus is to be made using following methods：
(1) inclined-plane culture：The slant medium prepared in following ratios：5~15g/L of yeast extract, 5~15g/L of glucose,
5~10g/L of peptone, 1~3g/L of potassium dihydrogen phosphate, tomato juice 30~50g/L, pH 6.5~7.0；Lactobacillus acidophilus is inoculated with
In the slant medium, under the conditions of 35~40 DEG C of cultivation temperature, 16~24h is cultivated, obtains lactobacillus acidophilus kind.
(2) first order seed culture：The primary-seed medium prepared in following ratios：5~15g/L of yeast extract, glucose 5
~15g/L, peptone 7.5g/L, potassium dihydrogen phosphate 2.0g/L, pH 7.0~7.5, by the lactobacillus acidophilus kind obtained by step (1)
It is inoculated in the primary-seed medium, under the conditions of 35~40 DEG C of cultivation temperature, cultivates 16~20h, obtain first order seed
Nutrient solution；
(3) secondary seed culture：The secondary seed medium prepared in following ratios：5~15g/L of yeast extract, glucose 5
~15g/L, peptone 7.5g/L, potassium dihydrogen phosphate 2.0g/L, Tween 80 0.5g/L, agar powder 20.0g/L, pH 7.0~
7.5, the first order seed nutrient solution obtained by step (2) is inoculated in the secondary seed medium, in cultivation temperature 35~40
Under the conditions of DEG C, 24~36h is cultivated, obtains secondary seed nutrient solution；
(4) fermented and cultured：The liquid fermentation medium prepared in following ratios：5~10g/L of peptone, yeast extract 3~
10g/L, 2~8g/L of tomato juice, 5~10g/L of glucose, 0.05~0.5g/L of magnesium sulfate, 1~3g/L of Triammonium citrate, acetic acid
Sodium 2~5g/L, pH6.8-7.2；With the inoculum concentration of liquid fermentation medium weight 6~10%, by the two level liquid obtained by step (3)
Body seed culture fluid is inoculated with the liquid fermentation medium, in 100~200r/min of speed of agitator, fermentation temperature 35~50
Under the conditions of DEG C, 20~30h of fermented and cultured, lactobacillus acidophilus fermentation culture is obtained；
(5) prepared by bacterium powder：Lactobacillus acidophilus fermentation culture obtained by step (4) is centrifuged and collects wet acidophilus breast
Bacillus body, after wet lactobacillus acidophilus body is mixed with mannitol protective agent, trehalose, add macropore starch carrier and adsorbed,
Crushed again after low temperature drying and lactobacillus acidophilus powder is made；Wherein, the weight of the mannitol protective agent and wet lactobacillus acidophilus body
Than for 0.15~0.5:1(g/Kg)；The weight ratio of the trehalose and wet lactobacillus acidophilus body is 5~20:1(g/Kg)；It is described
The weight ratio of macropore starch carrier and wet lactobacillus acidophilus body is 0.2~1:1(Kg/Kg).
Resist it is understood that lactobacillus acidophilus (Lactobacillusacidophilus) of the present invention can secrete
Biotin class material (acidolin, bacillus acidophilus's element, lactein), produces antagonism to pathogenic entero becteria, suppresses intestines
The propagation of road undesirable microorganism, adjustment gut flora balance, can assimilate amarogentin, cellobiose, aesculin, fructose, malt
Sugar, galactolipin, lactose, sucrose, mannose, melibiose, raffinose, trehalose, it is impossible to assimilate arabinose, gluconate,
Mannose.
In a preferred embodiment of this invention, the Lactobacillus plantarum 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；Lactobacillus plantarum kind is seeded on slant medium, Yu Wen
Under the conditions of 25~37 DEG C of degree, 24~48h is cultivated, obtains Lactobacillus plantarum kind；
(2) first order seed culture：Primary-seed medium is prepared in following ratios：5~15g/L of peptone, beef extract 5~
15g/L, 3~10g/L of yeast extract, 1~3g/L of diammonium hydrogen citrate, 1~3g/L of dipotassium hydrogen phosphate, Sodium acetate trihydrate 3~
8g/L, pH 6.2~6.4；Lactobacillus plantarum kind obtained by step (1) is inoculated with primary-seed medium, in temperature 25~37
DEG C, under the conditions of 150~300r/min of rotating speed, cultivate 18~30h, obtain level liquid seed culture fluid；
(3) secondary seed culture：Secondary seed medium is prepared in following ratios：5~15g/L of peptone, beef extract 5~
15g/L, 3~10g/L of yeast extract, 1~3g/L of diammonium hydrogen citrate, 1~3g/L of dipotassium hydrogen phosphate, Sodium acetate trihydrate 3~
8g/L, four 0.1~0.5g/L of water manganese sulfate, 0.2~1g/L of epsom salt, 15~30g/L of glucose, Tween 80 0.5~
2g/L, pH 6.2~6.4；Level liquid seed culture fluid obtained by step (2) is inoculated with secondary seed medium, in temperature
25~37 DEG C, under the conditions of 150~300r/min of rotating speed, cultivate 20~48h, obtain secondary liquid seed culture fluid；
(4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：5~10g/L of sucrose, 10~20g/ of peptone
L, 0.5~2g/L of potassium dihydrogen phosphate, 0.1~0.5g/L of dipotassium hydrogen phosphate, 0.1~0.5g/L of calcium chloride, ammonium sulfate 0.1~
0.5g/L, magnesium sulfate 0.05~0.2g/L, pH 7.2~7.5；, will with the inoculum concentration of liquid fermentation medium weight 6~10%
Secondary seed nutrient solution obtained by step (3) is seeded on the liquid fermentation medium, under the conditions of 30~35 DEG C, anaerobism hair
24~36h of ferment, obtain Lactobacillus plantarum fermentation culture；
(5) prepared by bacterium powder：Lactobacillus plantarum fermentation culture obtained by step (4) is centrifuged and collects wet plant breast
Bacillus body, after mannitol protective agent, Tween 80 are mixed with wet Lactobacillus plantarum body, add macropore starch carrier and adsorbed,
Crushed again after low temperature drying and Lactobacillus plantarum powder is made；Wherein, the weight of the mannitol protective agent and wet Lactobacillus plantarum body
Than for 0.15~0.5:1(g/Kg)；The weight ratio of the Tween 80 and wet Lactobacillus plantarum body is 5~20:1(g/Kg)；It is described
The weight ratio of macropore starch carrier and wet Lactobacillus plantarum body is 0.2~1:1(Kg/Kg).
It is understood that Lactobacillus plantarum of the present invention is one kind of lactic acid bacteria, optimum growth temperature is 30~
35, anaerobism or amphimicrobian, strain is single, sometimes in pairs or into chain, optimal pH 6.5 or so, category to be straight or curved shaft-like
In homofermentative lactic bacteria.Plant lactobacillus can produce lactic acid in enteron aisle by being metabolized, and suppress the growth of harmful bacteria and numerous
Grow, can repair the normal function of enteron aisle, maintain the enteral ecological balance, there is improvement enteron aisle, reduce diarrhoea, abdomen rises, constipation and other effects；
Plant lactobacillus has antibacterial action, can not only defend Listeria, staphylococcus and leukonid, moreover it is possible to resist Gram-negative
The invasion of the harmful bacterias such as bacterium, catalase-positive organism, Escherichia coli and salmonella.Lactobacillus plantarum is by reducing in serum
Cholesterol level have the function that reduce blood fat：Directly absorb the cholesterol in enteron aisle；Cholesterol is changed into polymer to sink
Form sediment and excrete；Produce the bio-enzyme degradation cholesterol of high activity.
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, 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 with and sent out into the liquid
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 rod-short, and no pod membrane, peritrichous, energy
The gram-positive bacterium of motion, it is caused withered grass element in bacillus subtilis growth course, polymyxins, nystatin, short
Bacitracin isoreactivity material has obvious inhibitory action to the conditioned pathogen of pathogenic bacteria or autogenous infection；Fish, shrimp intake are withered
After careless bacillus, bacillus subtilis consumes rapidly the free oxygen in environment, causes enteron aisle hypoxemia, promotes to give birth to beneficial to anaerobic bacteria
It is long, and the organic acids such as lactic acid are produced, gut pH is reduced, suppresses other pathogenic bacteria growings indirectly；Stimulating animal immune organ
Grow, activation T, bone-marrow-derived lymphocyte, improve immunoglobulin and antibody level, strengthen cellular immunity and humoral immunity work(
Can, improve herd immunity；The enzymes such as bacillus subtilis thalline itself synthesis alpha-amylase, protease, lipase, cellulase
Class, together played a role with the digestive enzymes in animal body in alimentary canal；Vitamin B can be synthesized1、B2、B6, nicotinic acid etc. it is more
Kind of B family vitamin, improve fish, the activity of interferon and macrophage in shrimp body.
In embodiments of the present invention, the bacillus licheniformis is to be made using following methods：
(1) inclined-plane culture：The slant medium prepared in following ratios：5~15g/L of peptone, yeast extract 3~
10g/L, 5~15g/L of sodium chloride, 0.5~2g/L of manganese sulfate, agar powder 10~20g/L, pH 7.2~7.4；By lichens gemma bar
Strain is seeded on slant medium, under the conditions of 35~40 DEG C of temperature, is cultivated 18~20h, is obtained bacillus licheniformis kind；
(2) first order seed culture：The primary-seed medium prepared in following ratios：5~15g/L of peptone, yeast carry
Take 3~10g/L of thing, 5~15g/L of sodium chloride, manganese sulfate 0.5~2g/L, pH 7.2~7.4；By the lichens bud obtained by step (1)
Spore bacillus specie is inoculated in primary-seed medium, under conditions of 30~40 DEG C, 100~400r/min, cultivates 18~20h,
Primary seed solution is made；
(3) secondary seed culture：The secondary seed medium prepared in following ratios：5~15g/L of peptone, yeast carry
Take 3~10g/L of thing, 5~15g/L of sodium chloride, manganese sulfate 0.5~2g/L, pH 7.2~7.4；By the one-level kind obtained by step (2)
Sub- liquid is inoculated in secondary seed medium, under conditions of 30~40 DEG C, 100~400r/min, is cultivated 20~36h, is made
Secondary seed solution；
(4) fermented and cultured：The liquid fermentation medium prepared in following ratios：20~30g/L of corn flour, beancake powder 5~
15g/L, 5~15g/L of glucose, 3~5g/L of urea, sodium chloride 5~15g/L, pH 7.0~7.5, with liquid fermentation medium
The inoculum concentration of weight 6~10%, the secondary liquid seed culture fluid obtained by step (3) is inoculated with into the liquid fermentation medium
In, under conditions of 100~200r/min of speed of agitator, 0.5~0.8vvm of throughput, 35~40 DEG C of fermentation temperature, fermentation training
20~30h is supported, obtains the lichen bacillus ferments nutrient solution；
(5) prepared by bacterium powder：The lichen bacillus ferments medium centrifugal obtained by step (4) is separated and collects wetland clothing
Bacillus body, after mannitol protective agent, Tween 80 are mixed with wet bacillus licheniformis body, add macropore starch carrier and carry out
Absorption, then crushed after low temperature drying and wet bacillus licheniformis powder is made；Wherein, the mannitol protective agent and wetland clothing gemma bar
The weight ratio of thalline is 0.15~0.5:1(g/Kg)；The weight ratio of the Tween 80 and wet bacillus licheniformis body is 5~20:1
(g/Kg)；The weight ratio of the macropore starch carrier and wet bacillus licheniformis body is 0.2~1:1(Kg/Kg).
It is understood that bacillus licheniformis (Bacillus Subtilis) cellular morphology of the present invention and row
Row are in shaft-like, Dan Sheng, and adjustable flora imbalance reaches therapeutic purposes, and body can be promoted to produce antibacterial substance, kill and cause a disease
Bacterium, after it enters fish, shrimp enteron aisle, by its unique biology take oxygen mechanism, to pathogenic bacteria such as staphylococcuses in enteron aisle
There is antagonism, and have promotion growth to beneficial bacteriums such as Bifidobacterium, Bacillus acidi lactici, bacteroid, digestibility streptococcus, oneself
Obtaining Minister Agriculture of China allows to add use in feed, is very outstanding feed industry thalline additive.
Second aspect, present invention also offers a kind of life of the raising fishes and shrimps intestinal health degree prepared as described in relation to the first aspect
The method of thing ferment, comprises the following steps：
The bacillus coagulans, lactobacillus acidophilus, Lactobacillus plantarum, bacillus subtilis, bacillus licheniformis are mixed
Composite bacteria agent is synthesized, then by after the composite bacteria agent of gained, synergist, activator, carrier mixing, is made and improves fishes and shrimps intestinal health
The biologic ferment of degree.
The third aspect, present invention also offers a kind of biological ferment of raising fishes and shrimps intestinal health degree as described in relation to the first aspect
Application of the element in feed is prepared.
Biologic ferment added with raising fishes and shrimps intestinal health degree as described in relation to the first aspect in the feed, the raising fish
The addition of the biologic ferment of shrimp intestinal health degree is 0.005~0.01 times of feed gross weight.
The preparation of the bacillus coagulans of embodiment 1
The bacillus coagulans of the embodiment of the present invention 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 with 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
(30 DEG C) crush bacillus coagulans powder is made afterwards；Wherein, the weight of the mannitol protective agent and wet bacillus coagulans body
Than for 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
The weight ratio of carrier and wet bacillus coagulans body is 0.6:1(Kg/Kg).
The preparation of the lactobacillus acidophilus of embodiment 2
The lactobacillus acidophilus of the embodiment of the present invention is to be made using following methods：
(1) inclined-plane culture：The slant medium prepared in following ratios：Yeast extract 10g/L, glucose 10g/L, peptone
8g/L, potassium dihydrogen phosphate 2g/L, tomato juice 40g/L, pH 6.8；Lactobacillus acidophilus is inoculated in the slant medium,
Under the conditions of 37 DEG C of cultivation temperature, 18h is cultivated, obtains lactobacillus acidophilus kind.
(2) first order seed culture：The primary-seed medium prepared in following ratios：Yeast extract 10g/L, glucose 10g/
L, peptone 7.5g/L, potassium dihydrogen phosphate 2.0g/L, pH 7.2, the lactobacillus acidophilus kind obtained by step (1) is inoculated in described
In primary-seed medium, under the conditions of 37 DEG C of cultivation temperature, 18h is cultivated, obtains first order seed nutrient solution；
(3) secondary seed culture：The secondary seed medium prepared in following ratios：Yeast extract 10g/L, glucose 10g/
L, peptone 7.5g/L, potassium dihydrogen phosphate 2.0g/L, Tween 80 0.5g/L, agar powder 20.0g/L, pH 7.4, by step (2)
The first order seed nutrient solution of gained is inoculated in the secondary seed medium, under the conditions of 37 DEG C of cultivation temperature, cultivates 28h,
Obtain secondary seed nutrient solution；
(4) fermented and cultured：The liquid fermentation medium prepared in following ratios：Peptone 7.5g/L, yeast extract 5g/L, kind
Tomato juice 6g/L, glucose 7.5g/L, magnesium sulfate 0.2g/L, Triammonium citrate 2g/L, sodium acetate 4g/L, pH 7.0；Sent out with liquid
The inoculum concentration of ferment culture medium weight 8%, the secondary liquid seed culture fluid obtained by step (3) is inoculated with and trained into the liquid fermentation
Support in base, under the conditions of speed of agitator 150r/min, 37 DEG C of fermentation temperature, fermented and cultured 24h, obtain lactobacillus acidophilus fermentation training
Nutrient solution；
(5) prepared by bacterium powder：Lactobacillus acidophilus fermentation culture obtained by step (4) is centrifuged and collects wet acidophilus breast
Bacillus body, after wet lactobacillus acidophilus body is mixed with mannitol protective agent, trehalose, add macropore starch carrier and adsorbed,
Low temperature drying (30 DEG C) crushes afterwards again is made lactobacillus acidophilus powder；Wherein, the mannitol protective agent and wet lactobacillus acidophilus body
Weight ratio be 0.2:1(g/Kg)；The weight ratio of the trehalose and wet lactobacillus acidophilus body is 10:1(g/Kg)；The macropore
The weight ratio of starch carrier and wet lactobacillus acidophilus body is 0.5:1(Kg/Kg).
The preparation of the Lactobacillus plantarum of embodiment 3
The Lactobacillus plantarum of the embodiment of the present invention is to be made using following methods：
(1) 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；Lactobacillus plantarum kind is seeded on slant medium, under the conditions of 28 DEG C of temperature, culture
36h, obtain Lactobacillus plantarum kind；
(2) first order seed culture：Primary-seed medium is prepared in following ratios：Peptone 10g/L, beef extract 10g/L,
Yeast extract 7.5g/L, diammonium hydrogen citrate 2g/L, dipotassium hydrogen phosphate 2g/L, Sodium acetate trihydrate 5g/L, pH 6.2；By step
(1) in the Lactobacillus plantarum kind inoculation primary-seed medium obtained by, under the conditions of 32 DEG C of temperature, rotating speed 200r/min, culture
24h, obtain level liquid seed culture fluid；
(3) secondary seed culture：Secondary seed medium is prepared in following ratios：Peptone 10g/L, beef extract 10g/L,
Yeast extract 7.5g/L, diammonium hydrogen citrate 2g/L, dipotassium hydrogen phosphate 2g/L, Sodium acetate trihydrate 6g/L, four water manganese sulfates
0.3g/L, epsom salt 0.5g/L, glucose 20g/L, Tween 80 1.5g/L, pH 6.4；By the one-level obtained by step (2)
In liquid seeds nutrient solution inoculation secondary seed medium, under the conditions of 37 DEG C of temperature, rotating speed 200r/min, 36h is cultivated, is obtained
Secondary liquid seed culture fluid；
(4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：Sucrose 10g/L, peptone 18g/L, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.65g/L, dipotassium hydrogen phosphate 0.35g/L, calcium chloride 0.25g/L, ammonium sulfate 0.3g/L, magnesium sulfate 0.1g/L, pH 7.2；
With the inoculum concentration of liquid fermentation medium weight 8%, the secondary seed nutrient solution obtained by step (3) is seeded to the liquid and sent out
On ferment culture medium, under the conditions of 32 DEG C, 24~36h of anaerobic fermentation, Lactobacillus plantarum fermentation culture is obtained；
(5) prepared by bacterium powder：Lactobacillus plantarum fermentation culture obtained by step (4) is centrifuged and collects wet plant breast
Bacillus body, after mannitol protective agent, Tween 80 are mixed with wet Lactobacillus plantarum body, add macropore starch carrier and adsorbed,
Low temperature drying (30 DEG C) crushes afterwards again is made Lactobacillus plantarum powder；Wherein, the mannitol protective agent and wet Lactobacillus plantarum body
Weight ratio be 0.2:1(g/Kg)；The weight ratio of the Tween 80 and wet Lactobacillus plantarum body is 8:1(g/Kg)；The macropore
The weight ratio of starch carrier and wet Lactobacillus plantarum body is 0.4:1(Kg/Kg).
The preparation of the bacillus subtilis of embodiment 4
The bacillus subtilis of the embodiment of the present invention is to be made using following methods：
(1) 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 temperature 28
30h is cultivated at DEG C, obtains bacillus subtilis strain；
(2) 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, rotating speed 200r/min, 30h is cultivated, obtains level liquid seed culture fluid；
(3) secondary seed culture：The secondary seed medium prepared in following ratios：Peptone 20g/L, beef extract 5g/
L, sodium chloride 5g/L, dipotassium hydrogen phosphate 1g/L, pH 7.0；The level liquid seed culture fluid of gained is seeded to the two level kind
In sub- culture medium, under the conditions of rotating speed 200r/min, throughput 0.8vvm, 28 DEG C of temperature, 32h is cultivated, obtains secondary liquid kind
Sub- nutrient solution；
(4) fermented and cultured：Liquid fermentation medium is prepared in following ratios：Peptone 20g/L, beef extract 5g/L, yeast
Cream 5g/L, potassium dihydrogen phosphate 1g/L, magnesium sulfate 0.5g/L, pH 7.4；, will with the inoculum concentration of liquid fermentation medium weight 8%
The secondary liquid seed culture fluid of gained is inoculated with 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；
(5) prepared by bacterium powder：The fermentation of bacillus subtilis medium centrifugal of gained is separated and collects wet bacillus subtilis
Thalline, after mannitol protective agent, Tween 80 are mixed with wet bacillus subtilis thalline, add macropore starch carrier and adsorbed,
Low temperature drying (30 DEG C) crushes afterwards again is made bacillus subtilis powder；Wherein, the mannitol protective agent and wet bacillus subtilis
The weight ratio of thalline is 0.2:1(g/Kg)；The weight ratio of the Tween 80 and wet bacillus subtilis thalline is 10:1(g/Kg)；Institute
The weight ratio for stating macropore starch carrier and wet bacillus subtilis thalline is 0.5:1(Kg/Kg).
The preparation of the bacillus licheniformis of embodiment 5
The bacillus licheniformis of the embodiment of the present invention is to be made using following methods：
(1) inclined-plane culture：The slant medium prepared in following ratios：Peptone 12g/L, yeast extract 7.5g/L,
Sodium chloride 10g/L, manganese sulfate 1g/L, agar powder 15g/L, pH 7.2；Bacillus licheniformis kind is seeded on slant medium,
18h is cultivated at 37 DEG C of temperature, obtains bacillus licheniformis kind；
(2) first order seed culture：The primary-seed medium prepared in following ratios：Peptone 10g/L, yeast extract
7.5g/L, sodium chloride 10g/L, manganese sulfate 1.5g/L, pH 7.4；Bacillus licheniformis kind obtained by step (1) is inoculated in one
In level seed culture medium, under conditions of 35 DEG C, 200r/min, 20h is cultivated, first order seed nutrient solution is made；
(3) secondary seed culture：The secondary seed medium prepared in following ratios：Peptone 10g/L, yeast extract
5g/L, 5~15g/L of sodium chloride, manganese sulfate 1.5g/L, pH 7.4；First order seed nutrient solution obtained by step (2) is inoculated in two
In level seed culture medium, under conditions of 37 DEG C, 200r/min, 28h is cultivated, secondary seed nutrient solution is made；
(4) fermented and cultured：The liquid fermentation medium prepared in following ratios：Corn flour 25g/L, beancake powder 10g/L, Portugal
Grape sugar 10g/L, urea 4g/L, sodium chloride 10g/L, pH 7.5, with the inoculum concentration of liquid fermentation medium weight 8%, by step
(3) the secondary seed nutrient solution obtained by is inoculated with the liquid fermentation medium, in speed of agitator 150r/min, throughput
Under the conditions of 0.7vvm, 37 DEG C of fermentation temperature, fermented and cultured 24h, the lichen bacillus ferments nutrient solution is obtained；
(5) prepared by bacterium powder：The lichen bacillus ferments medium centrifugal obtained by step (4) is separated and collects wetland clothing
Bacillus body, after mannitol protective agent, Tween 80 are mixed with wet bacillus licheniformis body, add macropore starch carrier and carry out
Absorption, then low temperature drying (30 DEG C) crush wet bacillus licheniformis powder is made afterwards；Wherein, the mannitol protective agent and wetland clothing
The weight ratio of bacillus body is 0.4:1(g/Kg)；The weight ratio of the trehalose and wet bacillus licheniformis body is 8:1(g/
Kg)；The weight ratio of the macropore starch carrier and wet bacillus licheniformis body is 0.8:1(Kg/Kg).
Embodiment 6~15
It is including as follows the embodiments of the invention provide a kind of preparation method for the biologic ferment for improving fishes and shrimps intestinal health degree
Step：
(1) bacillus coagulans, the embodiment of the present invention 2 of the gained of the embodiment of the present invention 1 are taken according to ratio as shown in table 1
The lactobacillus acidophilus of gained, the Lactobacillus plantarum of the gained of the embodiment of the present invention 3, the bacillus subtilis of the gained of the embodiment of the present invention 4
Bacterium, the bacillus licheniformis of the gained of the embodiment of the present invention 5, citric acid, vitamin E, plant ash, macroporous absorbent resin.
The raw materials used dosage statistics of 1 each embodiment of table
(2) by the bacillus coagulans of the gained of the embodiment of the present invention 1, the lactobacillus acidophilus of the gained of the embodiment of the present invention 2, sheet
The Lactobacillus plantarum of the gained of inventive embodiments 3, the bacillus subtilis of the gained of the embodiment of the present invention 4, the gained of the embodiment of the present invention 5
Bacillus licheniformis be mixed into composite bacteria agent, then the composite bacteria agent of gained, citric acid, vitamin E, plant ash, macropore are inhaled
After attached resin mixing, the biologic ferment for improving fishes and shrimps intestinal health degree is made, gained improves the biological ferment of fishes and shrimps intestinal health degree
Element total active bacteria number be respectively 4,100,000,000/gram, 5,600,000,000/gram, 6,200,000,000/gram, 5,800,000,000/gram, 5,700,000,000/gram, 5,400,000,000/
Gram, 6,300,000,000/gram, 4,800,000,000/gram, 4,200,000,000/gram.
Comparative example 1
This comparative example provides a kind of preparation method for the biologic ferment for improving fishes and shrimps intestinal health degree, including as follows
Step：
(1) raw material of following percentage by weights is taken：The lactobacillus acidophilus 10% of the gained of the embodiment of the present invention 2, the present invention are real
Apply the Lactobacillus plantarum 10% of the gained of example 3, the bacillus subtilis 10% of the gained of the embodiment of the present invention 4, the institute of the embodiment of the present invention 5
Bacillus licheniformis 10%, citric acid 2%, vitamin E 3%, plant ash 5%, macroporous absorbent resin 50%.
(2) by the lactobacillus acidophilus of the gained of the embodiment of the present invention 2, Lactobacillus plantarum, this hair of the gained of the embodiment of the present invention 3
Bacillus subtilis, the bacillus licheniformis of the gained of the embodiment of the present invention 5 of the bright gained of embodiment 4 are mixed into composite bacteria agent, then
After the composite bacteria agent of gained, citric acid, vitamin E, plant ash, macroporous absorbent resin are mixed, raising fishes and shrimps enteron aisle is made and is good for
Kang Du biologic ferment.
Comparative example 2
This comparative example provides a kind of preparation method for the biologic ferment for improving fishes and shrimps intestinal health degree, including as follows
Step：
(1) raw material of following percentage by weights is taken：The bacillus coagulans 20% of the gained of the embodiment of the present invention 1, the present invention
The lactobacillus acidophilus 10% of the gained of embodiment 2, the bacillus licheniformis 10% of the gained of the embodiment of the present invention 5, citric acid 2%, dimension
Raw plain E3%, plant ash 5%, macroporous absorbent resin 50%.
(2) by the bacillus coagulans of the gained of the embodiment of the present invention 1, the lactobacillus acidophilus of the gained of the embodiment of the present invention 2, sheet
The bacillus licheniformis of the gained of inventive embodiments 5 is mixed into composite bacteria agent, then composite bacteria agent, citric acid, vitamin by gained
E, after plant ash, macroporous absorbent resin mixing, the biologic ferment for improving fishes and shrimps intestinal health degree is made.
Comparative example 3
This comparative example provides a kind of preparation method for the biologic ferment for improving fishes and shrimps intestinal health degree, including as follows
Step：
(1) raw material of following percentage by weights is taken：The bacillus coagulans 20% of the gained of the embodiment of the present invention 1, the present invention
The Lactobacillus plantarum 10% of the gained of embodiment 3, the bacillus licheniformis 10% of the gained of the embodiment of the present invention 5, citric acid 2%, dimension
Raw plain E3%, plant ash 5%, macroporous absorbent resin 50%.
(2) by the bacillus coagulans of the gained of the embodiment of the present invention 1, the Lactobacillus plantarum of the gained of the embodiment of the present invention 3, sheet
The bacillus licheniformis of the gained of inventive embodiments 5 is mixed into composite bacteria agent, then composite bacteria agent, citric acid, vitamin by gained
E, after plant ash, macroporous absorbent resin mixing, the biologic ferment for improving fishes and shrimps intestinal health degree is made.
Illustrate the biologic ferment of the raising fishes and shrimps intestinal health degree of the present invention 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 area is about 2
Mu, 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, 3.0 are will be about
The Penaeus Vannmei seedling of centimetre body long size is launched in 12 mouthfuls of pond waters, and test group and control group are conventionally supported
Penaeus Vannmei is grown, unlike：In test group, by the biologic ferment of raising fishes and shrimps intestinal health degree provided by the invention with fitting
The water mixing of amount, add after commercially available honest shrimp feed is stirred, place fed again after obtained mixed fodder within 30 minutes to
(biologic ferment of fishes and shrimps intestinal health degree provided by the invention is commercially available honest shrimp feed weight in test group pond waters
0.01 times)；Control group only feeds Penaeus Vannmei using commercially available honest shrimp feed, and specific packet is as shown in table 2；During experiment
The health condition of (90 days) observation Penaeus Vannmeis, collects dead shrimp, and count Penaeus Vannmei from blowdown pipe sleeve net daily
Survival rate, the output value, statistical result is as shown in Fig. 1~2.
The experiment packet of the culture of Penaeus vannamei of table 2
| Packet | Feed |
| Test group 1 | The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 6 |
| Test group 2 | The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 7 |
| Test group 3 | The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 8 |
| Test group 4 | The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 9 |
| Test group 5 | The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 10 |
| Test group 6 | The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 11 |
| Test group 7 | The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 12 |
| Test group 8 | The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 13 |
| Test group 9 | The product of the commercially available gained of honest shrimp feed+embodiment of the present invention 14 |
| Test group 10 | The product of the gained of commercially available honest shrimp feed+comparative example of the present invention 1 |
| Test group 11 | The product of the gained of commercially available honest shrimp feed+comparative example of the present invention 2 |
| Test group 12 | 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 |
It will be seen from figure 1 that in test group 1~9 of the present invention the survival rate of Penaeus Vannmei more than 81%, wherein,
The survival rate highest of the Penaeus Vannmei of test group 2, reaches more than 85%, and Penaeus Vannmei is below in test group 10~12
74%, Penaeus Vannmei is only 52.3% in control group, the raising fishes and shrimps enteron aisle that this explanation embodiment of the present invention 6~14 is provided
The biologic ferment of health degree is advantageous to improve the immunity of Penaeus Vannmei.
Figure it is seen that in test group 1~9 of the present invention the output value of Penaeus Vannmei more than 530Kg/ mus, its
In, the output value highest of the Penaeus Vannmei of test group 2, reach more than 620Kg/ mus, and Penaeus Vannmei in test group 10~12
435Kg/ mus are below, Penaeus Vannmei is less than 300Kg/ mus in control group, what this explanation embodiment of the present invention 6~14 was provided
The biologic ferment for improving fishes and shrimps intestinal health degree is advantageous to improve the function of intestinal canal of Penaeus Vannmei, improves food conversion ratio, changes
The intestinal microecology balance of kind Penaeus Vannmei.
Total described, the biologic ferment for the raising fishes and shrimps intestinal health degree that the embodiment of the present invention 6~14 is provided is South America
The formation of the intestinal environment of white shrimp creates good condition, passes through aerobic bacteria bacillus subtilis and bacillus licheniformis
Interaction produces more nutriments and digestive ferment (such as amylase, protease), while can also exhaust Penaeus Vannmei intestines
Oxygen in road, good growing environment is created for the breeding of bacillus coagulans, Lactobacillus plantarum, lactobacillus acidophilus, is had
Beneficial to the immunity for improving Penaeus Vannmei；Be advantageous to improve the function of intestinal canal of Penaeus Vannmei, improve food conversion ratio, improve
The intestinal microecology balance of raw 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 area is about 2
Mu, is divided into 12 groups, wherein, 11 groups are test group, and one group is control group；According to the throw-in density of 10,000 tails/mu, by 100 tails/
The Tilapia mossambica fry of Kg specifications is launched in 12 mouthfuls of pond waters, and test group and control group conventionally cultivate Rofe
Fish, unlike：In test group, the biologic ferment of raising fishes and shrimps intestinal health degree provided by the invention is mixed with appropriate water
Close, add after commercially available honest fish meal is stirred, place 30 minutes and fed again after obtained mixed fodder to test group pond
(biologic ferment provided by the invention for improving fishes and shrimps intestinal health degree is the 0.005 of commercially available honest fish meal weight in the water body of the pool
Times)；Control group only feeds Tilapia mossambica using commercially available honest fish meal, and specific packet is as shown in table 3；See within (90 days) during experiment
The health condition of Tilapia mossambica is examined, collects dead Tilapia mossambica from blowdown pipe sleeve net daily, and counts the survival rate of Tilapia mossambica, the output value,
Statistical result is as shown in figs. 34.
The Tilapia mossambica culture experiment of table 3 is grouped
From figure 3, it can be seen that in test group 1~9 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 5, up to 92.5%, and Tilapia mossambica is not higher than 79% in test group 10~12, in control group
Tilapia mossambica is only 62.5%, the biologic ferment for the raising fishes and shrimps intestinal health degree that this explanation embodiment of the present invention 6~14 is provided
Be advantageous to improve the immunity of Tilapia mossambica.
From fig. 4, it can be seen that in test group 1~9 of the present invention the output value of Tilapia mossambica more than 4100Kg/ mus, wherein, examination
The survival rate highest of the Tilapia mossambica of group 5 is tested, is reached more than 4800Kg/ mus, and Tilapia mossambica is below in test group 10~12
3800Kg/ mus, Tilapia mossambica is less than 2500Kg/ mus in control group, the raising fishes and shrimps that this explanation embodiment of the present invention 6~14 is provided
The biologic ferment of intestinal health degree is advantageous to improve the function of intestinal canal of Tilapia mossambica, improves food conversion ratio, improves the intestines of Tilapia mossambica
Road microecological balance.
Total described, the biologic ferment for the raising fishes and shrimps intestinal health degree that the embodiment of the present invention 6~14 is provided is Rofe
The formation of the intestinal environment of fish creates good condition, mutual by aerobic bacteria bacillus subtilis and bacillus licheniformis
Effect produces more nutriments and digestive ferment (such as amylase, protease), while can also exhaust the oxygen in Tilapia mossambica enteron aisle
Gas, good growing environment is created for the breeding of bacillus coagulans, Lactobacillus plantarum, lactobacillus acidophilus, is advantageous to improve
The immunity of Tilapia mossambica；Be advantageous to improve the function of intestinal canal of Tilapia mossambica, improve food conversion ratio, improve the micro- life of enteron aisle of Tilapia mossambica
State balances.
The artificial viable bacteria infection experiment of effect example 3
The healthy Tilapia mossambica that initial weight is 150g or so is chosen, is randomly divided into 13 groups, 3 repetitions of every group of setting, each
Repetition has the tail of Tilapia mossambica 100, puts in a suitable place to breed in pond net box, and during experiment, test group improves fishes and shrimps enteron aisle by provided by the invention
The biologic ferment of health degree mixes with appropriate water, adds after commercially available honest fish meal is stirred, places 30 minutes and make
(the provided by the invention biologic ferment that improves fishes and shrimps intestinal health degree is fed into test group pond waters again after obtaining mixed fodder
For 0.005 times of commercially available honest fish meal weight)；Control group only feeds Tilapia mossambica using commercially available honest fish meal, specific point
Group is as shown in table 4；After experiment is carried out 30 days, the amplification of Escherichia coli viable bacteria is diluted to 1.5 × 105Cfu/ml, raised orally with mixing
Mode only give bacterium solution amount 0.2ml according to every tail Tilapia mossambica is single.It is small in 96 of Tilapia mossambica orally containing Escherichia coli viable bacteria feed
When it is interior, record the morbidity of each group Tilapia mossambica, death condition, as shown in table 4：
The each group Tilapia mossambica Experimental infection result of table 4
As shown in table 4, continuously feed and with the addition of raising fishes and shrimps intestinal health degree prepared by the embodiment of the present invention 6~14
Biologic ferment 30 days, Tilapia mossambica have significant preventive effect to enteritis caused by the Escherichia coli living bacterial liquid of orally administration, and nine
The death rate of individual test group Tilapia mossambica is not higher than 10%, wherein, the death rate of Tilapia mossambica is minimum in test group 5；And test group
The death rate of 10~12 Tilapia mossambicas is respectively 35%, 28.3%, 23%, after control group peroral infection Escherichia coli, 75.2%
Tilapia mossambica is lethal because of infection-induced hemorrhagic enteritis.This explanation, the raising fishes and shrimps described in inventive formulation are added in feed
The biologic ferment of intestinal health degree, it can effectively prevent the enteritis or lethal of Escherichia coli peroral infection initiation.
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)
- A kind of 1. biologic ferment for improving fishes and shrimps intestinal health degree, it is characterised in that：The life for improving fishes and shrimps intestinal health degree The raw material of thing ferment, by weight percentage, including：Bacillus coagulans 15~25%, lactobacillus acidophilus 5~15%, plant Lactobacillus 0~10%, bacillus subtilis 5~15%, bacillus licheniformis 0~15%, synergist 1~5%, activator 1~ 5%th, carrier 30 as one kind~55%.
- 2. the biologic ferment according to claim 1 for improving fishes and shrimps intestinal health degree, it is characterised in that：The raising fishes and shrimps Total active bacteria number of the biologic ferment of intestinal health degree is not less than 4,000,000,000/gram.
- 3. the biologic ferment according to claim 1 for improving fishes and shrimps intestinal health degree, it is characterised in that：The synergist choosing One or more from fumaric acid, lactic acid, citric acid, malic acid, tartaric acid, sorbic acid.
- 4. the biologic ferment according to claim 1 for improving fishes and shrimps intestinal health degree, it is characterised in that：The activator choosing From in amino acid, chromium picolinate, zinc sulfate, sodium selenite, vitamin E, calcium iodate, copper sulphate, ferrous sulfate, manganese sulfate It is one or more of.
- 5. the biologic ferment according to claim 1 for improving fishes and shrimps intestinal health degree, it is characterised in that：The carrier is selected from In plant ash, macropore starch, powder of straw, corn flour, rice bran, zeolite powder, precipitated calcium carbonate, wheat bran, flour, macroporous absorbent resin One or more.
- 6. the biologic ferment according to claim 5 for improving fishes and shrimps intestinal health degree, it is characterised in that：The carrier includes Plant ash, macroporous absorbent resin, wherein, the weight ratio of the plant ash and macroporous absorbent resin is 0.1~0.5:1.
- 7. the biologic ferment according to claim 1 for improving fishes and shrimps intestinal health degree, it is characterised in that：The raising fishes and shrimps The raw material of the biologic ferment of intestinal health degree, by weight percentage, including：Bacillus coagulans 20%, lactobacillus acidophilus 10%th, Lactobacillus plantarum 10%, bacillus subtilis 10%, bacillus licheniformis 10%, synergist 2%, activator 3%, load Body 35%.
- 8. the biologic ferment according to claim 1 for improving fishes and shrimps intestinal health degree, it is characterised in that：The raising fishes and shrimps The raw material of the biologic ferment of intestinal health degree, by weight percentage, including：Bacillus coagulans 21%, lactobacillus acidophilus 12%th, bacillus subtilis 12%, synergist 4%, activator 3%, carrier 48%.
- 9. a kind of method of the biologic ferment of the raising fishes and shrimps intestinal health degree prepared described in any one of claim 1~8, it is special Sign is：Comprise the following steps：The bacillus coagulans, lactobacillus acidophilus, Lactobacillus plantarum, bacillus subtilis, bacillus licheniformis are mixed into Composite bacteria agent, then by after the composite bacteria agent of gained, synergist, activator, carrier mixing, be made and improve fishes and shrimps intestinal health degree Biologic ferment.
- 10. a kind of biologic ferment of the raising fishes and shrimps intestinal health degree described in any one of claim 1~8 is preparing feed addition Application in agent.
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| CN110637948A (en) * | 2019-10-31 | 2020-01-03 | 江苏桂龙生物技术有限公司 | Livestock and poultry freshwater fish compound feed and preparation method thereof |
| CN112753849A (en) * | 2021-01-20 | 2021-05-07 | 南京众成正源生物技术有限公司 | Feed additive for improving immunity of fishes and shrimps as well as preparation method and application thereof |
| CN109231494B (en) * | 2018-10-29 | 2021-08-10 | 清远海贝生物技术有限公司 | Water quality improver for prawn culture and preparation method thereof |
| CN114223808A (en) * | 2021-12-09 | 2022-03-25 | 南京宝辉生物饲料有限公司 | Penaeus vannamei feed prepared from multiple strains and preparation method and application thereof |
| CN116998593A (en) * | 2023-09-15 | 2023-11-07 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Application of lactobacillus plantarum in preparation for improving fish sugar tolerance |
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