CN109287887A - A kind of feed of grass carp and preparation method thereof - Google Patents
A kind of feed of grass carp and preparation method thereof Download PDFInfo
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- CN109287887A CN109287887A CN201811123744.7A CN201811123744A CN109287887A CN 109287887 A CN109287887 A CN 109287887A CN 201811123744 A CN201811123744 A CN 201811123744A CN 109287887 A CN109287887 A CN 109287887A
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- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/02—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from meat
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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
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The present invention provides a kind of feed of grass carp, belongs to Aquatic Feed Processing technical field, includes the earthworm antimicrobial peptide hydrolyzate and active dry yeasr metabolin for accounting for feed granules weight 3.5 ~ 5.5% and 4.5 ~ 6.5% respectively in the feed；The present invention also provides the preparation methods of above-mentioned feed, earthworm antimicrobial peptide, active dry yeasr metabolin, solid carrier, water and additive are carried out secondary granulation by this method, it obtains containing after peptide particles, it is dry, it is coated with again with grease and polyethylene glycol, up to feed granules after drying, above-mentioned earthworm antimicrobial peptide preparation, which is used, contains 4- pyridine carboxaldehyde and rhamnolipid in buffer solution system.A kind of feed of grass carp provided by the present invention has the advantages that antibacterial, resisting stress, anti-mildew, good dispersion, can enhance immunity of organisms, improves fish intestines mucous membrane form, can long-term preservation, and directly reduce or substitute the use of conventional antibiotic；Provided preparation method production cost is low, short preparation period, and resource utilization and yield are high, can protect the high activity of beneficiating ingredient.
Description
Technical field
The invention belongs to Aquatic Feed Processing technical fields, and in particular to a kind of feed of grass carp and preparation method thereof.
Background technique
Grass carp belongs to Cypriniformes Cyprinidae Leuciscinae grass carp category.Grass carp has been commonly called as: grass carp, oily grass carp, careless grass carp, ctenopharyngodon idellus, grass
Fish, grass roots (northeast), charlatan, black black carp etc..Grass carp often inhabits the rivers and lakes of plains region, under general happiness occupy in water
Layer and the more water plant regions of offshore.Property it is active, swimming rapidly, often hunt in packs.For typical herbivorous fishes.In mainstream or lake
Deep water at it is overwintering.Breeding season parent population, which is traced back, swims habit.Grass carp fry and seed stage ingest zooplankter, midge children
Worm, Copepods and nauplius etc. switch to phytophage later to juvenile fish stage, rapidly because of its growth, eat in the natural environment
Object source is wide, is one of four large Chinese carps of CHINESE FRESHWATER cultivation.
With feed industry, aquaculture day to develop, it is also increasing to the demand of fish meal so that fish meal supply increasingly
Anxiety, price rise.And due to marine fishery resources, gradually failure, environmental protection and feed cost etc. are various,
Fish meal yield is reducing year by year, and cultivates demand and constantly rising, in aquaculture industry, fish meal use gradually by
To limitation.Vegetable protein sources are extensive, cheap, stable yield, are ideal substitution protein sources.But plant substitutes
Albumen contain it is some lose flavor substance and anti-nutritional factors, cause that palatability is poor, digests and absorbs the problems such as rate is low.Palatability
Difference directly results in aquatic livestock and ingests not positive or food refusal, and feed overlong time in water had both caused the waste of feed, while
Cause the pollution of breeding water body.DDGS (Distillers Dried Grains with Solubles, corn distiller's dried grain and
Soluble matter) it is the corn for being processed rear acquisition to poor liquid in the process using corn as main raw material(s) preparing ethanol by fermentation
Alcohol grain and raffinate dried object, not only protein rich in, amino acid, vitamin, fat, fermented distilled life in the process
At unknown growth factor, also rich in being conducive to the several mineral materials of growth of animal.Therefore, DDGS is as a kind of protein feed
Raw material can be widely used in Production of Livestock and Poultry.
However be not singly DDGS, biological feedstuff is in long-term storage, if mismanagement, easily by potential mould poison
The pollution such as element, aflatoxin, or adverse effect generated to animal health and production performance after feeding, or discarded mildew feed and
Wasting phenomenon and feed cost is caused to increase, and the fiber of high-content and non-starch polysaccharide will affect phosphorus in DDGS and vegetable protein
The absorption of equal mineral ions, the utilization rate for reducing nutriment and its poor palatability etc. limit the absorption of feed
It utilizes.Animal body is to that can generate adaptation reaction simultaneously when being stimulated by the variation of internal and external environment, and this reaction is by more
The stimulation of kind of factor and generates, including inherent inherent cause, Reproduction factor, the environmental factor in the external world etc., these stress be because
Plain " stressor " stimulation animal can all cause damages to animal body, and it is tired to show as animal spirit, appetite abolish, or extremely
Excitement, manic uneasiness, feeding are reduced；Anabolism weakens, and feed conversion rate reduces, and production performance decline, immunity weakens, mistake
The clinical symptoms such as quick.Therefore, reinforce feeding management while, in feed be added growth promotion, resisting stress, the resistance to bacterium of antibacterial activity
Substance directly can effectively adjust resisting stress, the disease resistance of cultivated animals, reduce the use of antibiotic.
The prior art such as Chinese invention patent CN1178693C discloses a kind of preparation method of earthworm antimicrobial peptide, pass through by
The centrifuge separation of earthworm extract that itself hydrolase of earthworm body resolves into, enters boiling water bath, ultrafiltration and etc. obtain earthworm antimicrobial peptide
Crude extract；Then by crude extract by molecular sieve column chromatography, ion-exchange chromatography, desalination and etc. purified to obtain earthworm
Earthworm antibacterial peptide, although to produce product purity higher for gained antibacterial peptide, production technology is complicated, unit titer production cost compared with
Height, and after multiple chromatography, the antibacterial activity of antibacterial peptide and the activity of digestive ferment can be reduced, to be added to feed meeting
Influence its using effect.
Summary of the invention
One of the objects of the present invention is to provide a kind of antibacterial, resisting stress, anti-mildew, good dispersion, enhancing immunity of organism
Power improves intestinal mucosa form, feed of grass carp with long preservation period.
The second object of the present invention be to provide a kind of short preparation period, production cost is low, resource utilization and yield are high,
Protect the preparation method of the feed of grass carp of beneficiating ingredient high activity.
The technical solution that the present invention is taken to achieve the above object are as follows:
Used yeast is saccharomyces cerevisiae in the present invention, is purchased from Changzhou Kai Qiao Biotechnology Co., Ltd.
A kind of preparation method of feed of grass carp, this method comprises: earthworm antimicrobial peptide, active dry yeasr metabolin, solid are carried
Body, water and additive carry out secondary granulation, obtain and contain peptide particles；It is dry above-mentioned containing peptide particles, then applied with grease and polyethylene glycol
Cloth is drying to obtain feed granules；Contain 4- pyridine carboxaldehyde and rhamnolipid in above-mentioned earthworm antimicrobial peptide preparation buffer solution system.
By earthworm antimicrobial peptide and in extreme circumstances, the active resisting stress product of yeast metabolism is added in feed the preparation method, Neng Gouyou
Effect ground improves mould proof toxin ability of the feed in long-term storage, transportational process, reduces feed mold loss, can also improve fish
Disease-resistant, resisting stress, the resistance to bacterium ability of antibacterial, and directly reduce or substitute conventional antibiotic use.
Preferably, application step are as follows: after containing peptide particles with grease spraying, then the polyethylene glycol for being 55~60% with concentration
Aqueous solution spraying.The anti-dust, anti-steam and water repelling property that feed granules can be increased using polyethylene glycol, help to protect
It deposits and transports, it helps rapid dispersion, the feed granules for avoiding grease from wrapping up are quickly dispersed in water because failing to feed in water
In and waste caused by cannot being preyed in time by grass carp.
Further preferably, the average molecular weight of polyethylene glycol is 1800~4400 dalton, and dosage is feed granules weight
The 25~30% of amount.The polyethylene glycol fusing point of the molecular weight is dry using lower temperature at 50~55 DEG C, therefore when drying, can be with
It protects the activity of antibacterial peptide and the active dry yeasr metabolin in particle not to be destroyed, maximum effect can be played.
Preferably, secondary granulation step are as follows: solid carrier is first carried out to 105~110 DEG C of quenched 10~20min of high temperature,
Then it pelletizes, obtains carrier granular；Carrier granular, earthworm antimicrobial peptide, active dry yeasr metabolin, water and additive are mixed again
Afterwards, 51~55 DEG C of quenched 5~10min of low temperature are carried out to get peptide particles are contained.First time high temperature is quenched to be made to contain in solid carrier
Starch sufficiently cure, while destroying the bioenzyme activity in carrier mass, be conducive to the long-term preservation of feed, second of low temperature
The quenched bioactivity that can retain antibacterial peptide, the enzyme preparation etc. that vitamin, metabolism generate.
Preferably, earthworm antimicrobial peptide is prepared by the following method: after taking earthworm to spit sand, clean, being placed in 2~3 times of amounts
In buffer, then it is passed through voltage stimulation 5~10min of earthworm of 5~10V, the coelomic fluid generated after electric shock is collected, then 2~8
DEG C, 25~35min is centrifuged under conditions of 1500~3000r/min, collect supernatant, and 5~100KDa is retained by ultrafiltration and is divided
The liquid of son amount to obtain the final product.A large amount of coelomic fluids can be quickly obtained using physical method stimulation earthworm, while decreasing chemical method
The adverse effect to protein in gained coelomic fluid is stimulated, it is a kind of that the hydrolyzate of gained antibacterial peptide, which has broad-spectrum antibacterial action,
It has no toxic side effect, noresidue, the substance without cause bacterial drug resistance, has growth promotion, health care and the function for treating disease to cultivated animals
Can, antibacterial peptide is added in feed, directly can reduce or substitute the use of conventional antibiotic.
Further preferably, in above-mentioned buffer solution system containing weight accounting be respectively 1~2% EDTA, 1~1.5%
2 mercapto ethanol, 0.15~0.23% 4- pyridine carboxaldehyde and 0.075~0.086% rhamnolipid, in buffer solution system
4- pyridine carboxaldehyde and rhamnolipid is added, can cooperate electric shock that earthworm is promoted to discharge more coelomic fluids, and reduce different liquid phases
Between surface tension, spread coelomic fluid quickly into buffer, thus reduce body cavity liquid enrichment generation inhibiting effect, obtain
More antibacterial proteins are taken, while the two can also destroy hydrogen bond balance relevant to N-H inside antibacterial protein together with electric current, subtract
The intermolecular and intramolecular of antibacterial protein conformation is maintained to conclude hydrogen bond, and then the structure of loose antibacterial protein less, in lock out operation
Middle antibacterial peptide is fast released and effectively catching, obtains higher resource utilization and yield.
Preferably, active dry yeasr metabolin is prepared by the following method: first by saccharomycete at 25~35 DEG C aerobic hair
9~10h of ferment, is subsequently placed in extreme environment 2~4h of anaerobic fermentation, acquisition stress metabolite, add 2.5~3.5wt%
Protease will be centrifuged from solution after promoting yeast cells self-dissolving 12~for 24 hours at 40~48 DEG C, take supernatant by 30000
The filter membrane of molecular weight, it is finally that filtrate is concentrated freeze-dried to obtain the final product.It, can rapid synthesis when stimulation of the yeast cells by external environment
Many have self-regeneration protective substances, to resist these bad conditions, as superoxide dismutase, glutathione,
These substances are added in feed in the protective factors such as vitamin C, vitamin E, can improve animal intestinal tract mucous membrane form, increase and raise
The utilization rate of material improves the anti-stress ability of animal, has the characteristics that feed safety, noresidue.
Further preferably, extreme environment includes extreme temperature, extreme oxidation or extreme osmotic pressure environment, specifically includes 4 ± 2
℃、Cl-Concentration 1 ± 0.5%, H2O20.2 ± 0.2mmol/L of concentration or 40 ± 2 DEG C, Cl-Concentration 1 ± 0.5%, H2O2Concentration 0.2
±0.2mmol/L.The metabolite that yeast generates in extreme circumstances, can increase cultivated animals low temperature, high temperature, it is hypertonic,
Adaptability under the external environments such as peroxidating keeps the metabolism and production performance of body, increases anti-stress ability.
The present invention also provides a kind of feed of grass carp, are made by the above method, and earthworm antimicrobial peptide and water are with earthworm in the feed
What the form of earthworm antibacterial peptide hydrolysate provided, above-mentioned earthworm antimicrobial peptide hydrolyzate accounts for the 3.5~5.5% of feed granules weight.It should
Feed has growth promotion, health care and the function for the treatment of disease to cultivated animals, can improve animal intestinal tract mucous membrane form, increases feed
Utilization rate, improve the immunity and anti-stress ability of animal, directly can reduce or substitute the use of conventional antibiotic.
Preferably, including the active dry yeasr metabolin and 2~3% that weight accounting is respectively 4.5~6.5% in feed
Additive, includes the nano-scale rare earth compound that weight accounting is 5~10% in above-mentioned additive, and above-mentioned rare earth compound is egg
At least one of propylhomoserin lanthanum, lanthanum oxalate, ceria.Rare earth can stimulate fish appetite, enhance digestive function, improve feed benefit
It is that a kind of stability is good, additive amount is small, the significantly excellent additive of gaining effect with rate.
It further preferably, further include multi-vitamins, composite mineral matter, choline chloride and glycine betaine in additive.
Preferably, the solid carrier in feed accounts for the 72~79% of feed granules weight, wherein with weight percent
Meter, including 2~15% fish meal, 35~45% insect powder, 13~18% fermentation rapeseed meal, 8~12% rice bran, 10~
15% starch and 5~20% corn protein powder.
Preferably, the grease in feed includes at least one of phosphatide oil, fish oil, soya-bean oil, lard, sheep oil.
The invention has the benefit that
1) antibacterial peptide is generated by using physical method stimulation earthworm in the present invention, short preparation period is at low cost, resource benefit
High with rate and yield, not vulnerable to the influence of external factor, made antibacterial peptide can enhance the immunity of organisms of aquatic livestock, have wide
Spectrum antibiotic property, has no drug resistance at noresidue, directly can reduce or substitute the use of conventional antibiotic；
2) in the present invention by carrying out extreme environment stimulation to yeast cells, yield is obtained more and the higher yeast of activity
Stress metabolite, by these metabolites be added feed in, animal intestinal tract mucous membrane form can be improved, increase the utilization of feed
Rate improves anti-stress ability of the animal under the external environments such as low temperature, high temperature, hypertonic, peroxidating, keeps the metabolism of body
And production performance；
3) height of the beneficiating ingredients such as active peptide in feed, enzyme preparation, vitamin is remained by secondary pelletize in the present invention
Activity, and be conducive to the anti-mildew and long-term preservation of feed, while comparing for extruding granulating process, it is greatly saved and is produced into
This；
4) feed granules are coated with using polyethylene glycol in the present invention, its anti-dust, anti-steam and water repelling property can be increased,
Facilitate storage and transport, the activity of antibacterial peptide and active dry yeasr metabolin can also be protected not to be destroyed in production, also
Help feed rapid dispersion in water, reduces waste and aquaculture cost.
Present invention employs above-mentioned technical proposals to provide a kind of feed of grass carp and preparation method thereof, compensates for the prior art
Deficiency, reasonable design, easy operation.
Specific embodiment
Technical solution of the present invention is described in further detail below in conjunction with specific embodiment:
Embodiment 1:
A kind of preparation method of feed of grass carp, this method comprises: earthworm antimicrobial peptide, active dry yeasr metabolin, solid are carried
Body, water and additive carry out secondary granulation, obtain and contain peptide particles；It is dry above-mentioned containing peptide particles, then applied with grease and polyethylene glycol
Cloth is drying to obtain feed granules；Contain 4- pyridine carboxaldehyde and rhamnolipid in above-mentioned earthworm antimicrobial peptide preparation buffer solution system.
By earthworm antimicrobial peptide and in extreme circumstances, the active resisting stress product of yeast metabolism is added in feed the preparation method, Neng Gouyou
Effect ground improves mould proof toxin ability of the feed in long-term storage, transportational process, reduces feed mold loss, can also improve fish
Disease-resistant, resisting stress, the resistance to bacterium ability of antibacterial, and directly reduce or substitute conventional antibiotic use.
Application step are as follows: after containing peptide particles with grease spraying, then the Aqueous Solutions of Polyethylene Glycol spraying for being 55% with concentration is
It can.The anti-dust, anti-steam and water repelling property that feed granules can be increased using polyethylene glycol, facilitate storage and transport,
Facilitate feed rapid dispersion in water, the feed granules for avoiding grease from wrapping up cannot be timely due to failing rapid dispersion in water
Waste caused by being preyed on by grass carp.
The average molecular weight of polyethylene glycol is 2200 dalton, and dosage is the 25% of feed granules weight.The molecular weight
Polyethylene glycol fusing point it is dry using lower temperature at 50~55 DEG C, therefore when drying, can protect antibacterial peptide in particle and
The activity of active dry yeasr metabolin is not destroyed, and can play maximum effect.
Secondary granulation step are as follows: solid carrier is first carried out to 105 DEG C of quenched 20min of high temperature, then pelletizes, obtains carrier
Grain；After again mixing carrier granular, earthworm antimicrobial peptide, active dry yeasr metabolin, water and additive, it is quenched to carry out 51 DEG C of low temperature
10min is to get containing peptide particles.First time high temperature is quenched to make the starch contained in solid carrier sufficiently cure, while destroying load
Bioenzyme activity in body substance is conducive to the long-term preservation of feed, and second of low temperature is quenched can to retain antibacterial peptide, dimension life
The bioactivity of the enzyme preparation that element, metabolism generate etc..
Earthworm antimicrobial peptide is prepared by the following method: after taking earthworm to spit sand, clean, being placed in the buffer of 2 times of amounts, then lead to
The voltage stimulation earthworm 10min for entering 5V, collects the coelomic fluid that generates after electric shock, then under conditions of 2 DEG C, 1500r/min from
Heart 35min collects supernatant, and retains the liquid of 5~100KDa molecular weight by ultrafiltration to obtain the final product.Earthworm is stimulated using physical method
Earthworm can be quickly obtained a large amount of coelomic fluids, while decrease chemical method stimulation to the bad shadow of protein in gained coelomic fluid
Ring, the hydrolyzate of gained antibacterial peptide has broad-spectrum antibacterial action, be one kind have no toxic side effect, noresidue, without causing bacterial drug resistance
Substance, to cultivated animals have growth promotion, health care and treat disease function, antibacterial peptide is added in feed, can directly be subtracted
Less or substitution conventional antibiotic use.
It containing weight accounting is respectively 1% EDTA, 1% 2 mercapto ethanol, 0.16% in above-mentioned buffer solution system
4- pyridine carboxaldehyde and 0.075% rhamnolipid, 4- pyridine carboxaldehyde and rhamnolipid are added in buffer solution system, can cooperate
Electric shock promotes earthworm to discharge more coelomic fluids, and reduces the surface tension between different liquid phases, makes coelomic fluid into buffer
Quickly diffusion obtains more antibacterial proteins, while the two can also be together with electricity to reduce the inhibiting effect of body cavity liquid enrichment generation
Stream destroys hydrogen bond balance relevant to N-H inside antibacterial protein together, reduces the intermolecular and molecule for maintaining antibacterial protein conformation
Interior conclusion hydrogen bond, and then the structure of loose antibacterial protein, antibacterial peptide is fast released simultaneously effectively catching in lock out operation, obtains
Higher resource utilization and yield.
Active dry yeasr metabolin is prepared by the following method: first by saccharomycete at 28 DEG C aerobic fermentation 10h, be subsequently placed in
Extreme environment anaerobic fermentation 2h, acquisition stress metabolite, add the protease of 2.5wt%, promote yeast at 42 DEG C
It after aqtocytolysis 20h, will be centrifuged from solution, supernatant is taken to pass through the filter membrane of 30000 molecular weight, it is finally that filtrate is concentrated freeze-dried i.e.
, above-mentioned culture medium is potato dextrose agar, pol 5%.When stimulation of the yeast cells by external environment
When, energy rapid synthesis many has the protective substance of self-regeneration, to resist these bad conditions, such as superoxide dismutase
The protective factors such as enzyme, glutathione, vitamin C, vitamin E are added these substances in feed, can improve animal intestinal tract mucous membrane
Form increases the utilization rate of feed, improves the anti-stress ability of animal, has the characteristics that feed safety, noresidue.
Extreme environment includes extreme temperature, extreme oxidation or extreme osmotic pressure environment, and specific packet is in 4 DEG C, Cl-Concentration 1%,
H2O2Ferment 1h under conditions of concentration 0.2mmol/L, is subsequently placed in 40 DEG C, Cl-Concentration 1%, H2O2The condition of concentration 0.2mmol/L
Lower fermentation 1h.The metabolite that yeast generates in extreme circumstances can increase cultivated animals in low temperature, high temperature, hypertonic, peroxide
Adaptability under the external environments such as change keeps the metabolism and production performance of body, increases anti-stress ability.
A kind of feed of grass carp prepared by the above method, earthworm antimicrobial peptide and water are hydrolyzed with earthworm antimicrobial peptide in the feed
What the form of liquid provided, earthworm antimicrobial peptide hydrolyzate accounts for the 3.5% of feed granules weight.The feed has growth-promoting to cultivated animals
Long, health care and the function for the treatment of disease, can improve animal intestinal tract mucous membrane form, increase the utilization rate of feed, improve animal
Immunity and anti-stress ability directly can reduce or substitute the use of conventional antibiotic.
In feed include weight accounting be respectively 4.5% active dry yeasr metabolin and 2% additive, above-mentioned additive
In include weight accounting be 5% nano-scale rare earth compound, above-mentioned rare earth compound be methionine lanthanum, lanthanum oxalate, titanium dioxide
The arbitrary proportion mixture of cerium.Rare earth can stimulate fish appetite, enhance digestive function, improve efficiency of feed utilization, be a kind of stabilization
Property is good, additive amount is small, the significantly excellent additive of gaining effect.
It in additive by weight percentage, further include 40% multi-vitamins, 40% composite mineral matter, 8%
Choline chloride and 10% glycine betaine.
Solid carrier in feed accounts for the 72% of feed granules weight, wherein by weight percentage, including 12% fish
Powder, 38% insect powder, 17% fermentation rapeseed meal, 11% rice bran, 15% starch and 7% corn protein powder.
The grease that weight accounting is 18% in feed is the arbitrary proportion mixture of phosphatide oil, soya-bean oil.
Embodiment 2:
A kind of feed of grass carp, by weight percentage, including 5.3% earthworm antimicrobial peptide hydrolyzate, 5.4% active ferment
Female metabolin, 2.5% additive, 78.5% solid carrier and 8.3% grease；Including 7.5% in above-mentioned additive
Methionine lanthanum, 38% multi-vitamins, 38% composite mineral matter, 10.5% choline chloride and 6% glycine betaine；It is above-mentioned
In solid carrier include 9.5% fish meal, 40% insect powder, 15% fermentation rapeseed meal, 8% rice bran, 12% starch and
15.5% corn protein powder；Above-mentioned grease is phosphatide oil.
A kind of preparation method of feed of grass carp, specifically includes the following steps:
1) it after cleaning earthworm, is placed in the buffer of 3 times of amounts, then is passed through the voltage stimulation earthworm 5min of 8V, collect electricity
Then the coelomic fluid generated after hitting is centrifuged 25min under conditions of 2 DEG C, 3000r/min, collect supernatant, and cut by ultrafiltration
It stays the liquid of 5~100KDa molecular weight to get earthworm antimicrobial peptide hydrolyzate, distinguishes in above-mentioned buffer solution system containing weight accounting
For 1.5% EDTA, 1% 2 mercapto ethanol, 0.21% 4- pyridine carboxaldehyde and 0.085% rhamnolipid；
2) first by saccharomycete at 30 DEG C aerobic fermentation 9.5h, be subsequently placed in extreme environment anaerobic fermentation 4h, acquisition is answered
Swash metabolite, adds the protease of 3wt%, after promoting yeast cells self-dissolving 18h at 48 DEG C, will be centrifuged, and take from solution
Supernatant passes through the filter membrane of 30000 molecular weight, finally that filtrate is concentrated freeze-dried up to active dry yeasr metabolin, above-mentioned extreme environment
It is specifically included in 2 DEG C, Cl-Concentration 1.5%, H2O2Ferment 2h under conditions of concentration 0.3mmol/L, then in 42 DEG C, Cl-Concentration
1.5%, H2O2Ferment 2h under conditions of concentration 0.3mmol/L, and above-mentioned culture medium is potato dextrose agar, pol
It is 10%；
3) solid carrier is first carried out to 110 DEG C of quenched 15min of high temperature, then pelletizes, obtains carrier granular；Again by carrier
After grain, earthworm antimicrobial peptide, active dry yeasr metabolin, water and additive mixing, 53 DEG C of quenched 8min of low temperature are carried out to get containing peptide
Grain；
4) after containing peptide particles with grease spraying, then the Aqueous Solutions of Polyethylene Glycol spraying for being 57% with concentration can obtain feed
Grain, the average molecular weight of above-mentioned polyethylene glycol are 4400 dalton, and dosage is the 29% of feed granules weight.
Embodiment 3:
A kind of feed of grass carp, by weight percentage, including 4.8% earthworm antimicrobial peptide hydrolyzate, 6.3% active ferment
Female metabolin, 3% additive, 75.5% solid carrier and 10.4% grease；It include 3.5% egg in above-mentioned additive
Propylhomoserin lanthanum, 4.5% lanthanum oxalate, 37% multi-vitamins, 37% composite mineral matter, 9.5% choline chloride and 8.5%
Glycine betaine；In above-mentioned solid carrier include 3% fish meal, 43% insect powder, 16% fermentation rapeseed meal, 9% rice bran,
11% starch and 18% corn protein powder；Above-mentioned grease is the arbitrary proportion mixture of phosphatide oil, soya-bean oil.
A kind of preparation method of feed of grass carp, specifically includes the following steps:
1) it after cleaning earthworm, is placed in the buffer of 3 times of amounts, then is passed through the voltage stimulation earthworm 8min of 6V, collect electricity
Then the coelomic fluid generated after hitting is centrifuged 30min under conditions of 4 DEG C, 3000r/min, collect supernatant, and cut by ultrafiltration
It stays the liquid of 5~100KDa molecular weight to get earthworm antimicrobial peptide hydrolyzate, distinguishes in above-mentioned buffer solution system containing weight accounting
For 1.8% EDTA, 1.5% 2 mercapto ethanol, 0.18% 4- pyridine carboxaldehyde and 0.079% rhamnolipid；
2) first by saccharomycete at 33 DEG C aerobic fermentation 9h, be subsequently placed in extreme environment anaerobic fermentation 3h, acquisition stress
Metabolite adds the protease of 3.5wt%, after promoting yeast cells self-dissolving 14h at 44 DEG C, will be centrifuged, and take from solution
Supernatant passes through the filter membrane of 30000 molecular weight, finally that filtrate is concentrated freeze-dried up to active dry yeasr metabolin, above-mentioned extreme environment
It is specifically included in 5 DEG C, Cl-Concentration 1.5%, H2O2Ferment 1.5h under conditions of concentration 0.2mmol/L, then in 41 DEG C, Cl-It is dense
Spend 1.5%, H2O2Ferment 1.5h under conditions of concentration 0.2mmol/L, and above-mentioned culture medium is potato dextrose agar,
Pol is 8%；
3) solid carrier is first carried out to 110 DEG C of quenched 10min of high temperature, then pelletizes, obtains carrier granular；Again by carrier
After grain, earthworm antimicrobial peptide, active dry yeasr metabolin, water and additive mixing, 52 DEG C of quenched 5min of low temperature are carried out to get containing peptide
Grain；
4) after containing peptide particles with grease spraying, then the Aqueous Solutions of Polyethylene Glycol spraying for being 60% with concentration can obtain feed
Grain, the average molecular weight of above-mentioned polyethylene glycol are 3600 dalton, and dosage is the 28% of feed granules weight.
Embodiment 4:
A kind of preparation method of feed of grass carp, wherein being optimized to the preparation of active dry yeasr metabolin, specific optimization step
It is rapid as follows: first by saccharomycete at 33 DEG C aerobic fermentation 9h, be subsequently placed in extreme environment anaerobic fermentation 3h, acquisition stress be metabolized
Product adds the protease of 3.5wt%, after promoting yeast cells self-dissolving 14h at 44 DEG C, will be centrifuged from solution, and take supernatant
Liquid passes through the filter membrane of 30000 molecular weight, and finally that filtrate is concentrated freeze-dried up to active dry yeasr metabolin, above-mentioned extreme environment is specific
It is included in 5 DEG C, Cl-Concentration 1.5%, H2O2Ferment 1.5h under conditions of concentration 0.2mmol/L, then in 41 DEG C, Cl-Concentration
1.5%, H2O2Ferment 1.5h under conditions of concentration 0.2mmol/L, and above-mentioned culture medium is potato dextrose agar, sugar
Degree is 8%, and the dihydroxyacetone (DHA) of 0.043wt% and the alpha-terpineol of 0.084wt%, the addition of the two are added when prepared by culture medium
The extension that can promote Yeast Growth logarithmic phase obtains more saccharomycete and its metabolite, and prior effect is to carry out
When extreme environment stimulates the stress reaction of yeast, especially when osmotic pressure changes, ion gradient on cell membrane is quickly adjusted
Variation, accelerate cell membrane on turgescence balance, prevent cell in moisture it is excessive loss or entrance, increased with this and extended carefully
The survival ability of born of the same parents, enables yeast sufficiently to discharge stress factor and biological enzyme, into protection mechanism, and then be metabolized yeast stress
Products collection efficiency gets a promotion, and keeps greater activity.
The present embodiment is optimized on the basis of embodiment 3, made feed of grass carp raw material and other preparation steps
It is consistent with embodiment 3, feed of grass carp is made.
Comparative example:
A kind of preparation method of feed of grass carp, wherein earthworm antimicrobial peptide is prepared by the following method: after earthworm is cleaned, being set
In the buffer that 3 times are measured, then it is passed through the voltage stimulation earthworm 8min of 6V, the coelomic fluid generated after electric shock is collected, then 4
DEG C, 30min is centrifuged under conditions of 3000r/min, collect supernatant, and by the liquid of ultrafiltration retention 5~100KDa molecular weight,
Up to earthworm antimicrobial peptide hydrolyzate, containing 1.8% EDTA, 1.5% 2 mercapto ethanol in above-mentioned buffer solution system, i.e., this is slow
When prepared by fliud flushing system, it is not added with 4- pyridine carboxaldehyde and rhamnolipid.
This comparative example is that test is compared on the basis of embodiment 3, made feed of grass carp raw material and other preparations
Step and embodiment 3 are consistent, and feed of grass carp is made.
Embodiment 5:
Feed of grass carp performance measurement
Using the made feed of Examples 1 to 4 as test group 1~4, the made feed of comparative example as a control group, takes equal amount
Feed granules be measured, measurement result such as the following table 1.
The test of 1 feed of grass carp basic performance of table
| Test group 1 | Test group 2 | Test group 3 | Test group 4 | Control group | |
| Antibacterial peptide content/% | 1.25 | 1.28 | 1.33 | 1.32 | 1.03 |
| Polypeptide total content/% | 8.45 | 8.63 | 8.79 | 10.22 | 7.95 |
| Organic acid content/% | 12.54 | 12.75 | 12.42 | 13.77 | 10.45 |
| Enzyme preparation content/% | 3.75 | 3.84 | 3.87 | 4.04 | 3.54 |
It can be obtained by upper table 1, antibacterial peptide content, polypeptide total content are substantially less than each test group in feed obtained by control group, say
Bright 4- pyridine carboxaldehyde and rhamnolipid are obviously improved effect to the yield of antibacterial peptide, also improve polypeptide in feed and always contain
Amount, while the polypeptide total content of test group 4, organic acid content and enzyme preparation content are apparently higher than other test groups, are due to examination
Testing group 4 joined dihydroxyacetone (DHA) and alpha-terpineol, and the two has the yield and activity of active dry yeasr metabolite to be increased significantly
Beneficial effect.
Embodiment 6:
Feed of grass carp feeding experiment
In certain fish culture production base, using the made feed of Examples 1 to 4 as test group 1~4, the made feeding of comparative example
Material as a control group, using certain commercially available careless fish feed as blank group, randomly chooses 12 pools mouthful, be divided into 2 groups it is parallel, put in a suitable place to breed identical
After quantity, original body mass are without the Grass Carp Juveniles after significant difference, domestication, feed day 3~4 times, daily ration of feeding be with the 3 of weight~
5%, 1 weight gain amount is estimated every 2 weeks, and adjusts feeding volume, during which not administration of antibiotics.
During cultivation, under the conditions of controlling the heat stress that whole temperature is 35~37 DEG C, after cultivation 72 days, acquisition sample point
Analysis, as a result such as the following table 2,3.
Influence of the feed of grass carp to Growth of Grass Carps Ctenopharyngodon Idellus performance under 2 Heat-temperature stress of table
| Test group 1 | Test group 2 | Test group 3 | Test group 4 | Control group | Blank group | |
| First counterpoise/g | 79.95 | 80.35 | 79.35 | 79.90 | 79.85 | 79.46 |
| Last counterpoise/g | 253.15 | 252.64 | 257.36 | 265.04 | 234.52 | 203.96 |
| Disease incidence/% | 6.7 | 6.8 | 6.7 | 5.9 | 7.5 | 11.5 |
| Survival rate/% | 98.5 | 98.2 | 98.1 | 99.0 | 97.1 | 92.8 |
| Coefficient of condition/% | 1.76 | 1.74 | 1.79 | 1.76 | 1.77 | 1.68 |
The Intestinal Tract Morphology index of 3 grass carp of table
| Fold quantity | Fold length | Fold width | Muscle layer thickness | |
| Test group 1 | 44.38 | 293.35 | 115.22 | 77.59 |
| Test group 2 | 44.63 | 287.56 | 116.72 | 78.89 |
| Test group 3 | 44.48 | 318.60 | 118.24 | 78.39 |
| Test group 4 | 46.23 | 341.10 | 122.57 | 82.48 |
| Control group | 42.33 | 273.20 | 104.57 | 76.54 |
| Blank group | 42.00 | 275.63 | 105.23 | 72.51 |
By upper table 2 it is found that under high temperature stimulation, blank group grass carp most terminal counterpoise be lower than test group and control group, be by
Resisting stress factor content is too low in blank group feed, causes grass carp under high temperature environment, is all not so good as to intake, the digestion of feed
Test group, so blank group end counterpoise is far below test group；Simultaneously hot environment and do not use antibiotic under conditions of, blank
The disease incidence of group is much higher than test group, and blank group survival rate is far below test group, is anti-due to what is added in test group feed
Bacterium peptide and active dry yeasr metabolin all have the function of antibacterial, resisting stress, therefore test group feeding effect is substantially better than blank group.
By upper table 3 it is found that control group and blank group are not much different in Intestinal Morphology indices, illustrate feeds utilized to reach
The nutritional need needed to grass carp normal growth, but the indices of test group are significantly superior in control group and blank group, work as pleat
For wrinkle when quantity increases, fold length and width becomes larger, animal is also stronger to the abilities of digestive and absorption of nutriment, and muscle layer is thicker,
Instestinal motility is more sufficient, also promotes abilities of digestive and absorption, so the grass carp of test group is substantially better than blank group on last counterpoise.
The prior art of routine techniques dawn known to those skilled in the art in above-described embodiment, therefore herein no longer in detail
It repeats.
The above embodiments are only used to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this field
Member can also make a variety of changes and modification without departing from the spirit and scope of the present invention.Therefore, all equivalent
Technical solution also belong to scope of the invention, scope of patent protection of the invention should be defined by the claims.
Claims (10)
1. a kind of preparation method of feed of grass carp, it is characterised in that: including,
Earthworm antimicrobial peptide, active dry yeasr metabolin, solid carrier, water and additive are subjected to secondary granulation, obtains and contains peptide particles；
It is dry described containing peptide particles, then be coated with grease and polyethylene glycol, it is drying to obtain feed granules；
Contain 4- pyridine carboxaldehyde and rhamnolipid in the earthworm antimicrobial peptide preparation buffer solution system.
2. a kind of preparation method of feed of grass carp according to claim 1, it is characterised in that: the application step are as follows: use
After grease spraying contains peptide particles, then the Aqueous Solutions of Polyethylene Glycol for being 55 ~ 60% with concentration sprays.
3. a kind of preparation method of feed of grass carp according to claim 1 or 2, it is characterised in that: the polyethylene glycol
Average molecular weight is 1800 ~ 4400 dalton, and dosage is the 25 ~ 30% of feed granules weight.
4. a kind of preparation method of feed of grass carp according to claim 1, it is characterised in that: the secondary granulation step
Are as follows: solid carrier is first carried out to 105 ~ 110 DEG C of quenched 10 ~ 20min of high temperature, then pelletizes, obtains carrier granular；Again by carrier
Grain, earthworm antimicrobial peptide, active dry yeasr metabolin, water and additive mixing after, carry out 51 ~ 55 DEG C of quenched 5 ~ 10min of low temperature to get
Containing peptide particles.
5. a kind of preparation method of feed of grass carp according to claim 1, it is characterised in that: the earthworm antimicrobial peptide passes through
Following methods preparation: earthworm being cleaned and is placed in buffer, then is passed through voltage stimulation 5 ~ 10min of earthworm of 5 ~ 10V, collects electricity
Then the coelomic fluid generated after hitting is centrifuged 25 ~ 35min under conditions of 2 ~ 8 DEG C, 1500 ~ 3000r/min, collect supernatant, and
Retain the liquid of 5 ~ 100KDa molecular weight by ultrafiltration to obtain the final product.
6. a kind of preparation method of feed of grass carp according to claim 1 or 5, it is characterised in that: the buffer solution system
In containing weight accounting be respectively 0.15 ~ 0.23% 4- pyridine carboxaldehyde and 0.075 ~ 0.086% rhamnolipid.
7. a kind of preparation method of feed of grass carp according to claim 1, it is characterised in that: the active dry yeasr metabolin
It is prepared by the following method: first by saccharomycete 9 ~ 10h of aerobic fermentation, being subsequently placed in extreme environment 2 ~ 4h of anaerobic fermentation, then plus
Enter after protease promotes yeast cells self-dissolving 12 ~ for 24 hours, will be centrifuged from solution, supernatant is taken to pass through the filter membrane of 30000 molecular weight,
It is finally that filtrate is concentrated freeze-dried to obtain the final product.
8. a kind of preparation method of feed of grass carp according to claim 7, it is characterised in that: the extreme environment includes pole
Temperature, extreme oxidation or extreme osmotic pressure environment are held, 4 ± 2 DEG C, Cl are specifically included-Concentration 1 ± 0.5%, H2O2Concentration 0.2 ±
0.2mmol/L or 40 ± 2 DEG C, Cl-Concentration 1 ± 0.5%, H2O20.2 ± 0.2mmol/L of concentration.
9. a kind of feed of grass carp as made from the preparation method of any one of claim 1 ~ 8, by it is characterized by: the feed
Middle earthworm antimicrobial peptide and water are provided in the form of earthworm antimicrobial peptide hydrolyzate；The earthworm antimicrobial peptide hydrolyzate accounts for feed
The 3.5 ~ 5.5% of grain weight.
10. a kind of feed of grass carp according to claim 9, it is characterised in that: include that weight accounting is distinguished in the feed
For 4.5 ~ 6.5% active dry yeasr metabolin and 2 ~ 3% additive；It include weight accounting in the additive is 5 ~ 10% to receive
Meter level rare earth compound；The rare earth compound is at least one of methionine lanthanum, lanthanum oxalate, ceria.
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| CN114790428A (en) * | 2022-05-20 | 2022-07-26 | 中国农业科学院饲料研究所 | Predigested feed for improving health and growth of carnivorous aquatic animals |
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