CN109845899A - A kind of sugarcane end pin white wine residue protein feed and preparation method thereof - Google Patents
A kind of sugarcane end pin white wine residue protein feed and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of sugarcane end pin white wine residue protein feeds and preparation method thereof, which is prepared by sugarcane end pin, white wine slag, molasses, corn flour, dregs of beans, wheat bran, rice bran and composite fermentation microbial inoculum.Domestication bacterium is from the Nature used in the present invention, by enrichment, screening, domestication obtains, the ability of decomposition of cellulose with super strength, obtained composite fermentation microbial inoculum, fermentation is efficient, stability is good, sugarcane end pin can be effectively reduced, the crude fiber content of the raw materials such as white wine slag, by crude fibre, the polymeric carbohydrate that the animals such as lignin are difficult to sufficiently digest and assimilate is converted to the absorbable lower-molecular substance utilized, improve the digestive utilization ratio of animal, obtained fermented fodder nutrient is more comprehensive, feed conversion rate is high, and the content of crude protein in fermented feed is improved simultaneously, improve the trophism and palatability of feed, it can avoid the use of antibiotic in fermentation process, safety and environmental protection.
Description
Technical field
The present invention relates to protein feed processing technique field, specifically a kind of sugarcane end pin white wine residue protein feed and its system
Preparation Method.
Background technique
The current situation of Chinese plant-eating animal is made a general survey of, the raising of China plant-eating animal is concentrated mainly on the north, especially western
Backlands area, but these areas had already appeared overgraze, vegetation degeneration the phenomenon that；Nowadays country encourages south to develop again,
Although the gross area of Grassland in south China is many, meadow in flakes is few, to form scale and herds cultivation and is difficult, this is also so
For many years, one of the factor of southern plant-eating animal development is restricted always.Want to form large-scale cultivation in south, it is necessary to big
The feed of amount.With the fast development of Chinese aquaculture, the demand day of animal feed more increases, both at home and abroad to cultivation albumen
The research of matter feed is increasingly deep, and protein feed has been largely used in the production of all kinds of mixed feeds, and achieves and largely grind
Study carefully achievement.The protein content of protein feed is high, full of nutrition, and content of ashes is high, and calcium phosphorus is abundant, and ratio is good, is conducive to
Raising animal is absorbed and utilized, stimulating growth and breeding to some extent, is that other nutriments institute is irreplaceable.
Sugarcane (Saccharum of ficinarum L.) be a kind of perennial grass family (Gram ineae) chinese sorghum race
High stalk monocotyledon, tropical and subtropical region is planted extensively.Sugarcane belongs to C4 plant, has very high conversion of solar energy,
Yield per unit area is higher, is a kind of industrial crops for efficiently utilizing solar energy.Contain sufficient moisture content, sugar, albumen in sugarcane
Winter-spring season can be effectively relieved as a kind of important non-grain fodder crop in matter, fat, organic acid and various vitamins etc.
Save the problem of feed scarcity.Guangxi is sugarcane main producing region, there is sugar industry by-product abundant such as bagasse, sugarcane toppers leaf, molasses
Deng these by-products have the characteristics that source is wide, quantity is big low with price, small by seasonal effect, as can these sugar industry by-products
Object fodder not only can solve the difficult situation that people and animals strive grain, considerable economic well-being of workers and staff also brought for sugar refinery.
Sugarcane is the important economical crops in Guangxi, and sugar industry is one of the pillar industry in Guangxi, and sugarcane end pin is sugarcane
The general designation that upper 2-3 tender sections and dark green blade are pushed up when cutting is received, weight is about the 10% of sugarcane weight, is the byproduct of Sugarcane Industry, contains
Sugar, protein, the various vitamins such as more than 20 kinds of amino acid, thiamine, riboflavin and vitamin B6, niacin, folic acid, pantothenic acid.
Fiber children is tender, sweet, agreeable to the taste, and cattle and sheep like to eat.Long-term practice proves that sugarcane end pin feeds ox, has growth promoting, laying on meat, lactation amount lasting
Growth and moistening lung to arrest cough, health care and other effects.The Sugarcane Industry in Guangxi provides estimable large for southern Winter-Spring withered grass season
Green forage resource, but sugarcane end pin is often got rid of discarded everywhere, and burn-up of setting on fire after it is dried naturally, utilization rate is very low, answers
Feeding main points are grasped according to its characteristic, are made full use of, to promote the development of plant-eating animal, and can be to solve the problems, such as that grain mentions
For a feasible exploitation route.
Summary of the invention
The object of the present invention is to provide a kind of sugarcane end pin white wine residue protein feed and preparation method thereof, the protein feed by
Sugarcane end pin, white wine slag, molasses, corn flour, dregs of beans, wheat bran, rice bran and composite fermentation microbial inoculum are prepared；It can be effectively reduced
The animals such as crude fibre, lignin, are difficult to the high score sufficiently digested and assimilated by the crude fiber content of the raw materials such as sugarcane end pin, white wine slag
Sub- carbohydrate is converted to the absorbable lower-molecular substance utilized, improves the digestive utilization ratio of animal, and obtained fermentation is raised
Material nutrient is more comprehensive, and feed conversion rate is high, and improves the content of crude protein in fermented feed simultaneously, improves the trophism of feed
And palatability, it can avoid the use of antibiotic, safety and environmental protection in fermentation process.
The invention is realized by the following technical scheme:
A kind of sugarcane end pin white wine residue protein feed, is prepared from the following raw materials in parts by weight: 70~90 parts of sugarcane end pin, white wine
It is 35~45 parts of slag, 20~30 parts of molasses, 25~35 parts of corn flour, 8~13 parts of dregs of beans, 8~13 parts of wheat bran, 8~13 parts of rice bran, multiple
Close 0.2~0.4 part of fermenting agent；
The white wine slag be using cereal or potato grain as raw material brewing white spirit after remaining residue；
The composite fermentation microbial inoculum is prepared from the following raw materials in parts by weight: 15~30 parts of bacterium freeze-dried powder of domestication, Candida utilis
8~10 parts of yeast freeze-dried powder, 6~8 parts of lactobacillus plantarum freeze-dried powder, 8~10 parts of bacillus subtilis freeze-dried powder, aspergillus niger freeze-drying
4~6 parts of powder, 4~6 parts of aspergillus oryzae freeze-dried powder, 7~10 parts of Trichoderma viride freeze-dried powder, 18~20 parts of cellulase freeze-dried powder.
The composite fermentation microbial inoculum the preparation method comprises the following steps:
(1) domestication bacterium is taken to expand the bacterium solution for reaching stationary phase when culture, centrifugation takes bacterium mud to be washed with sterile phosphate buffer,
Centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out vacuum freeze drying to get domestication bacterium freeze dried powder；
(2) candida utili is inoculated in malt extract medium, is cultivated under the conditions of 36~38 DEG C, pH6.3~6.5 to steady
Periodically, it takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, centrifugation is resuspended in sterile again
Embedding medium is added in phosphate buffer, carries out vacuum freeze drying to get candida utili freeze dried powder；
(3) lactobacillus plantarum is inoculated in MRS culture medium, culture takes to stationary phase under the conditions of 36~38 DEG C, pH6.3~6.5
Bacterium solution is centrifuged, and liquid is discarded supernatant, and bacterium mud is taken to be washed with sterile phosphate buffer, and it is slow that centrifugation is resuspended in sterile phosphate again
Embedding medium is added in fliud flushing, carries out vacuum freeze drying to get lactobacillus plantarum freeze dried powder；
(4) each bacterium branch is inoculated in PCA fluid nutrient medium, is cultivated under the conditions of 36~38 DEG C, pH6.8~7.2 to stationary phase,
It takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, centrifugation is resuspended in sterile phosphate again
Buffer, be added embedding medium, carry out vacuum freeze drying, that is, respectively obtain bacillus subtilis freeze-dried powder, aspergillus niger freeze-dried powder,
Aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulase freeze-dried powder.
(5) each freeze-dried powder obtained above is uniformly mixed to get composite fermentation microbial inoculum.
The MRS culture medium constituent are as follows: 8~10 parts of peptone, 8~10 parts of beef extract, 4~6 parts of yeast extract, lemon
1.5~2.2 parts of diammonium of lemon acid hydrogen, 18~22 parts of glucose, 0.8~1.2 part of Tween 80,4~6 parts of sodium acetate, dipotassium hydrogen phosphate
1.5~2.5 parts, 900~1000 parts of distilled water；The PCA fluid nutrient medium, constituent are as follows: 4~6 parts of tryptone, ferment
2~3.2 parts of female cream, 1~2 part of glucose, 950~1000 parts of distilled water.
The embedding medium is prepared from the following raw materials in parts by weight: 8~10 parts of maltodextrin, 6~8 parts of beta-cyclodextrin,
10~12 parts of starch, 8~20 parts of water.
The vacuum freeze drying parameter are as follows: 65 DEG C of condenser temperature ﹣, 30~35 DEG C of heating temperature, vacuum degree≤60pa,
The dry moisture content to microbial inoculum is no more than 7%.
The sugarcane end pin white wine residue protein feed, preparation method includes the following steps:
(1) pretreatment of raw material: sugarcane end pin is crushed to granularity no more than 5mm, drying to moisture content is spare no more than 12%；
(2) inoculation fermentation: corn flour, dregs of beans, wheat bran, rice bran are mixed in proportion, and molasses and water is added, and are adjusted moisture and are contained
Amount is inoculated with composite fermentation microbial inoculum, barrelling is sealed by fermentation 5~10 days to get fermentation fine fodder to 45~60%；
(3) it is sealed by fermentation: sugarcane end pin and molasses being added in fermentation fine fodder, is pressed and sealed and continues fermentation 20~24 days,
Middle fermentation fine fodder, sugarcane end pin, molasses three weight ratio be 1:3:1, fermentation is to there is sour slightly wine flavour to get sweet
Sugarcane end pin white wine residue protein feed.
It is described domestication bacterium acclimation method the following steps are included:
(1) it chooses bacterium mud: choosing the half a year above sugarcane field soil or sugarcane top stack retting mud or sugar refinery bagasse laydown area is stacked
Half a year sludge formed above seals at original bacterium mud scene in sterile glass vials up for safekeeping to get original bacterium mud, takes back laboratory work
It is cultivated for original flora；These original bacterium muds, due to the influence of ecological environment, gradually form in long-term stacking environment
Dominant bacteria, what is survived is the bacterium of decomposable by-product from sugarcane cellulose mostly, and such bacterium is easier to be tamed,
The ability of degraded cellulose can significantly improve, and have relative stability.
(2) primary domestication: taking fresh sugarcane end pin to be crushed to 100~200 mesh, in 121 DEG C of high pressure sterilizations 10~
20min is added to prepared basal medium and is uniformly mixed after cooling, the weight ratio of sugarcane end pin and basal medium is
1.0~1.5:1 is to get primary domestication culture medium；
According to the difference of sugarcane end pin additional amount, 5~6 gradients are set, prepare different primary tame and docile of several sugarcane end pin contents
Change culture medium, the original bacterium mud in step (1) is inoculated into each culture medium, 35~40 DEG C, cultivate under the conditions of pH6.3~6.5
20~30 days, the situation in 2~4 days one subcultures of record was decomposed up to excellent with sugarcane tail, clear with bacterium solution
Clearly, it is excellent, in comprehensive each culture medium situation that bacterium mud is not smelly, selects one group of optimal culture medium, survives in bacterium solution
Bacterium is primary domestication bacterium；
(3) secondary domestication: taking fresh sugarcane end pin to be crushed to 100~200 mesh, in 121 DEG C of 10~20min of high pressure sterilization, obtains A
Material；It takes the bagasse powder after juicing to be broken to 100~200 mesh, in 121 DEG C of 10~20min of high pressure sterilization, obtains B material；It takes in sugar refinery
The sewage that bagasse shower water is formed filters off sundries, obtains C material；
Above-mentioned A, B, C material are subjected to combination of two or three's combination respectively, are added to basal medium, is made multiple secondary tame and docile
Change culture medium, once tames the bacterium solution of bacterium in inoculation step (2) respectively, cultivated 15~30 days under the conditions of 35~42 DEG C, every 2
Situation in~4 days one subcultures of record, decomposes up to excellent, with clarifying contaminated liquids, not smelly with sugarcane tail, bagasse
It is excellent, in comprehensive each culture medium situation, selects one group of optimal culture medium, bacterium solution is secondary domestication bacterium；
The basal medium is prepared from the following raw materials in parts by weight: 8~10 parts of peptone, 8~10 parts of beef extract, and yeast
4~6 parts of cream, 5~10 parts of brewer's wort, 1.5~2.2 parts of diammonium hydrogen citrate, 18~22 parts of glucose, Tween 80 0.8~1.2
Part, 4~6 parts of sodium acetate, 1.5~2.5 parts of dipotassium hydrogen phosphate, 4~6 parts of tryptone, 2~3.2 parts of yeast extract, glucose 1~2
Part, 900~1000 parts of distilled water.
(4) expand culture: the bacterium solution for the secondary domestication bacterium for taking step (3) to obtain expands culture, and culture medium is secondary
Culture medium when culture medium optimal situation is tamed, 40~50h is cultivated；
(5) it dispenses, the sub- liquid of domesticated strain can be carried out continuing to expand culture, be formed a large amount of by preservation to get the sub- liquid of domesticated strain
Dominant bacteria, to adapt to industrialized production.
After domestication, the ability of decomposition of cellulose greatly promotes domestication bacterium of the present invention, especially decomposes
The ability of bagasse and sugarcane end pin, the survival ability in by-product from sugarcane are also improved；Tame bacterium it is main at
Dividing includes: candida utili, lactic acid bacteria, bacillus subtilis, aspergillus niger, aspergillus oryzae and Trichoderma viride.
Sugarcane end pin white wine residue protein feed applicable object of the present invention be the ruminants such as ox, sheep and chicken,
The poultry such as duck, goose.
Protein feed refers to natural moisture content lower than 45%, and crude fibre is lower than 18% feed in dry matter.According to main
Source is different, and protein feed can be divided into Plant protein feed, animal protein feed, single-cell protein feed and non-protein
Four major class of nitrogen feed.Physical property protein feed content is high, full of nutrition, conducive to being absorbed and utilized for raising animal, to some extent
Stimulating growth and breeding are that other nutriments institute is irreplaceable.
The general designation of upper 2-3 tender sections and dark green blade is pushed up when sugarcane end pin is sugarcane harvesting, weight is about sugarcane weight
10%, it is the byproduct of Sugarcane Industry, contains sugar, protein, more than 20 kinds of amino acid, thiamine, riboflavin and vitamin B6, cigarette
The various vitamins such as acid, folic acid, pantothenic acid.Fiber children is tender, sweet, agreeable to the taste, and cattle and sheep like to eat.Long-term practice proves that sugarcane end pin is fed
Ox has growth promoting, laying on meat, lactation amount sustainable growth and moistening lung to arrest cough, health care and other effects.
White wine slag is also known as vinasse, be using cereal or potato grain as raw material brewing white spirit after remaining residue.
Vinasse are the direct leftover bits and pieces during wine brewing, it, which not only contains a certain proportion of grain, can save the fine fodder for feeding ox, it is also
Crude protein rich in can reach 25% or so, about be higher by 2-3 times of corn content, at the same also containing various trace elements,
Vitamin, saccharomycete etc., the content of lysine, methionine and tryptophan is also very high, this is that agricultural crop straw cannot provide,
It is one as the feed of ox or other animals for it to select well.It is formed after all fermented rear thermophilic digestion of vinasse,
So the characteristics of its crude fiber content is lower, therefore vinasse have good palatability and be easy digestion as the feed of ox, and
And it can also effectively prevent Niu Fasheng bloat.White wine slag can not only improve absorptivity and palatability after everfermentation, also have
Improve the effect of animal appetite.
Dregs of beans is a kind of byproduct obtained after soybean extracting bean oil, also known as " Soybean Meal ".Dregs of beans albumen rich in
Matter and a variety of amino acid, gross protein value are up to 30~50%, are the primary protein sources of animal protein feed.It is unprocessed
Dregs of beans in containing antitrypsin, uremic enzyme, saponin, goitre incitant etc., to the digestion benefit of animal and feed
With can generate adverse effect, and dregs of beans of the invention can remove these adverse effects after the fermentation of special microbial inoculum, also
The palatability of feed can be improved, improve digestibility, the activity with unique physiological active functions is generated largely after dregs of beans is fermented
Peptide, easy to digest, absorb that fast, antigenicity is low, the proliferation of beneficial bacterium in effective stimulus enteron aisle adjusts the knot of internal Tiny ecosystem flora
Structure increases entire alimentary canal to the decomposition of feed nutrition substance, synthesis, absorption and utilization.
Wheat bran is also known as wheat bran, is the outermost epidermis of wheat, after wheat is machined by flour milling, becomes flour and two, wheat bran
Point, wheat bran is that wheat processing processes the byproduct obtained after flour.Wheat bran dietary fiber rich in and a variety of nutriments,
The absorption of animal body is more advantageous to after fermented.
Rice bran is the principal by product of paddy processing, mainly by pericarp, kind skin, perisperm, aleurone and embryo processing system
At generally can be divided into feed grade rice bran and two kinds of food-grade rice bran.Various nutrients and physiological activator are rich in rice bran,
Protein content average out to 15-18%, fatty 16-22%, rice bran is economical as ruminant and nutritive value feed abundant is former
Material, is main fine fodder supplement.
Composite fermentation microbial inoculum used in the present invention, active constituent include candida utili, lactobacillus plantarum, withered grass bud
Spore bacillus, aspergillus niger, aspergillus oryzae and Trichoderma viride, by the soil near sugarcane field soil, sugarcane top stack retting or sugar refinery
In each strain be enriched with, screened and expanded culture respectively, then add embedding medium carry out embedding treatment, carry out vacuum refrigeration
It is dry, finally microbial inoculum powder is mixed to form according to specific ratio.Each mutually coordinated cooperation of strain, fermentation is efficient, speed
Fastly, consume energy low, high conversion rate, and stability is good, can make up the deficiency of single culture fermentation, can also solve spontaneous fermentation strain
Excessively complicated, fermentation process controllability difference problem.Fermenting agent of the invention during the fermentation, by microorganism and its
The a variety of physiological metabolism substances generated, can effectively kill various pathogens, carry out to some harmful substances original in feed
Degradation and harmless processing, avoid the use of antibiotic.
Candida utili (Candida utilIs Torula utilis Henneb or edible torula) are called.Its protein and Wei Sheng
The content of plain B is all higher than brewer's yeast, it using urea and nitric acid as nitrogen source can not need that any growth is added in the medium
The factor can be grown.It can utilize pentose and hexose, not produce alcohol under aerobic conditions, can utilize the sulfurous of paper industry
Acid waste liquid, moreover it is possible to produce the edible protein of people and animals using molasses, wood hydrolysis liquid etc..The cell of candida utili is in
Round, ellipse or sausage shape, size are (3.5~4.5) μ m (7~13) μm.Liquid Culture does not produce mould, and tube bottom has thallus heavy
It forms sediment.In wort agar medium, bacterium colony milky, smoothly, and with or without gloss, neat in edge or mycelioid.Candida utilis
Animal intestinal micro-ecology balance is adjusted in yeast, improves feed digestibility, enhances and is rich in vitamin B in its cell of animal body power
And protein, moreover it is possible to part nutriment needed for animal is provided.
Lactobacillus plantarum (Lactobacillus plantarum) be lactic acid bacteria one kind, nose circle straight-bar bacterium, usually
0.9~1.2vtm × 3.0~8.0 μm, anaerobism or amphimicrobian, strain be it is straight or curved rod-shaped, individually, sometimes in pairs or chaining
Shape, optimal pH 6.5 or so, belongs to homofermentative lactic bacteria.Compared with this bacterium is its viable count with the difference of other lactic acid bacterias
Height can largely produce acid, increase the pH value stabilization in water not, and the acidic materials of its output can degrade heavy metal；It can hair
Ferment pentose or gluconate, 85% or more is lactic acid in final product；The energy distinctive lactobacillin of output in reproductive process,
Lactobacillin is a kind of preservative of bion, has certain antisepsis.
Bacillus subtilis (Bacillus subtilis) be bacillus one kind, individual cells 0.7~0.8 × 2
~3 microns, aerobic bacteria.Bacterium colony rough surface is opaque, dirty white or yellowish, when growing in liquid medium, is commonly formed
Wrinkle mould.Available protein, a variety of sugar and starch, decompose tryptophan and form indoles.During bacillus subtilis thalli growth
Subtilin, polymyxins, nystatin, the gramicidins isoreactivity substance of generation, these active materials are to pathogenic bacteria or interior
The conditioned pathogen of source sexuality dye has apparent inhibiting effect.
Aspergillus niger (Aspergillus niger), Deuteromycotina, Hyphomycetes, hyphomycetales, Moniliaceae, aspergillus is true
A Common Species in bacterium.It is distributed widely in grain all over the world, plant product and soil, is agriculturally used as production sugar
Change the strain of feed.Aspergillus niger is important fermentation industry strain, can produce amylase, acid protease, cellulase, pectin
Enzyme, glucose oxidase, citric acid, gluconic acid and gallic acid etc..
Aspergillus oryzae (Aspergillus oryzae (Asp.oryzae)) category Deuteromycotina, Hyphomycetes, hyphomycetales, from
Geng Bao section, a Common Species in aspergillus fungi, 150~300 μm of diameter.Aspergillus oryzae can produce protease, amylase, saccharification
Enzyme, cellulase, phytase etc..Under the action of amylase, straight chain, the amylopectin in raw material are degraded to dextrin and various
Low molecule carbohydrate, such as maltose, glucose；Under the action of protease, stodgy macro-molecular protein is degraded to
Peptone, polypeptide and various amino acid, and the difficult mass degradation absorbed such as can make crude fibre in auxiliary material, phytic acid, improve battalion
Value, health-care efficacy and digestibility are supported, the fermentation industries such as food, feed, production kojic acid, wine brewing are widely used in.
Trichoderma viride (Trichoderma viride) be Trichoderma one kind, it is extensive in distributed in nature, often it is saprophytic in
On timber, seed and plant residue.Trichoderma glues spore mushroom in Deuteromycotina, Hyphomycetes, hyphomycetales.Trichoderma viride energy
Generate one of a variety of biologically active enzyme systems, the highest bacterial strain of institute's cellulase-producing activity, generated cellulase pair
Crop has degradation.
Cellulase (cellulase), also known as β-Isosorbide-5-Nitrae-glucan -4- glucan hydrolysis, is that degraded cellulose generates Portugal
The general name of one group of enzyme of grape sugar, is a kind of complex enzyme, mainly by circumscribed 1,4 beta-glucanase, Endo-β-glucanase and β-glucose
The composition such as glycosides enzyme, there are also the zytases of very high vigor.Biocatalysis from decomposition of cellulose, can be by cellulose point
Solution is mainly used in feed, alcohol, weaving and food etc. at oligosaccharides or the protein of monosaccharide.
Beneficial effects of the present invention:
1, sugarcane end pin white wine residue protein feed of the present invention is the preparation using sugarcane end pin and white wine slag as primary raw material
At the protein feed that the ruminants such as suitable ox, sheep eat, it is inexpensive, without secondary pollution to realize sugarcane end pin, white wine slag
It comprehensively utilizes, is not generated " three wastes " in production process, it is environmentally friendly；Sugarcane end pin contains sugar, protein, amino acid and sulphur
The various vitamins such as amine element, riboflavin and vitamin B6, niacin, folic acid, pantothenic acid, through its knot can be effectively improved after everfermentation
Structure reduces crude fiber content, improves the digestive utilization ratio of ruminant, while microbial cells can also be promoted to degrade and utilize
Efficiency it is high, which to be processed into its nutritive value after feed, and corn flour, dregs of beans, wheat bran, rice bran is added etc. improves feed protein
Content, feed palatability is good, and for ruminants such as rear cattle, sheep, weight gain is fast, and animal is with good meat quality, and fiber finer, color are fresh, have
Toughness.
2, the composite fermentation microbial inoculum used in the present invention, active constituent include domestication bacterium, candida utili, plant cream bar
Bacterium, bacillus subtilis, aspergillus niger, aspergillus oryzae and Trichoderma viride, addition embedding medium carry out embedding treatment, it is dry to carry out vacuum refrigeration
It is dry, finally microbial inoculum powder is mixed to form according to specific ratio.Each mutually coordinated cooperation of strain, efficiently, speed is fast for fermentation,
It consumes energy low, high conversion rate, stability is good, can make up the deficiency of single culture fermentation, can also solve spontaneous fermentation strain mistake
In complicated, fermentation process controllability difference problem；The fermentation time of feed, obtained protein feed nutrition can also be shortened
More comprehensively, feed conversion rate is high.
3, during livestock and poultry feeding large-scale development, the long-term drug resistance that antibiotic is used continuously and causes animal body, thus
So that mankind pathogen is also generated drug resistance, endangers the health of the mankind.Fermenting agent of the invention during the fermentation, passes through micro- life
Object and its a variety of physiological metabolism substances of generation, can effectively kill various pathogens, to some nuisances original in raw material
Matter has carried out degradation and harmless processing, avoids the use of antibiotic, safety and environmental protection.
4, sugarcane end pin and white wine slag are the by-products of cane sugar factory, are not all fully utilized, make all the time
At a large amount of wastes of resource.The present invention carries out recycling using sugarcane end pin and white wine slag as primary raw material, by by-product from sugarcane
It utilizes, turns waste into wealth, the situation that people and animals strive grain can be alleviated, considerable economic well-being of workers and staff can be brought for sugar refinery, moreover it is possible to promote cattle and sheep
The development of aquaculture reduces aquaculture cost, has good economic benefit.
5, the domestication bacterium used in the present invention obtains from the Nature by enrichment, screening, domestication, with super strength
Breeding and adaptability, can pointedly decompose the cellulose in the by-product from sugarcane such as bagasse, sugarcane end pin, can be by thick fibre
The polymeric carbohydrate that the animals such as dimension, lignin are difficult to sufficiently digest and assimilate is converted to the absorbable low-molecular material utilized
Matter, and the content of crude protein in fermented feed is improved simultaneously, improve the trophism and palatability of feed.
Specific embodiment
In order to keep technical solution of the present invention and advantage clearer, below with reference to the embodiment of the present invention, to the present invention
Technical solution be clearly and completely described.
Embodiment 1
A kind of sugarcane end pin white wine residue protein feed, is prepared from the following raw materials in parts by weight: 70 parts of sugarcane end pin, white wine slag 45
Part, 20 parts of molasses, 25 parts of corn flour, 8 parts of dregs of beans, 13 parts of wheat bran, 13 parts of rice bran, 0.2 part of composite fermentation microbial inoculum；
The white wine slag be using sorghum as raw material brewing white spirit after remaining residue；
The composite fermentation microbial inoculum is prepared from the following raw materials in parts by weight: 15 parts of bacterium freeze-dried powder of domestication, candida utili
10 parts of freeze-dried powder, 8 parts of lactobacillus plantarum freeze-dried powder, 10 parts of bacillus subtilis freeze-dried powder, 6 parts of aspergillus niger freeze-dried powder, aspergillus oryzae
6 parts of freeze-dried powder, 10 parts of Trichoderma viride freeze-dried powder, 18 parts of cellulase freeze-dried powder.
The composite fermentation microbial inoculum the preparation method comprises the following steps:
(1) domestication bacterium is taken to expand the bacterium solution for reaching stationary phase when culture, centrifugation takes bacterium mud to be washed with sterile phosphate buffer,
Centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out vacuum freeze drying to get domestication bacterium freeze dried powder；
(2) candida utili is inoculated in malt extract medium, culture takes bacterium to stationary phase under the conditions of 37 DEG C, pH6.4
Liquid is centrifuged, and liquid is discarded supernatant, and bacterium mud is taken to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again
Embedding medium is added in liquid, carries out vacuum freeze drying to get candida utili freeze dried powder；
(3) lactobacillus plantarum is inoculated in MRS culture medium, culture takes bacterium solution to carry out to stationary phase under the conditions of 37 DEG C, pH6.4
Centrifugation, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again, adds
Enter embedding medium, carries out vacuum freeze drying to get lactobacillus plantarum freeze dried powder；
(4) each bacterium branch is inoculated in PCA fluid nutrient medium, under the conditions of 37 DEG C, pH6.9 culture to stationary phase, take bacterium solution into
Row centrifugation, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again,
Embedding medium is added, carries out vacuum freeze drying, that is, respectively obtains bacillus subtilis freeze-dried powder, aspergillus niger freeze-dried powder, aspergillus oryzae
Freeze-dried powder, Trichoderma viride freeze-dried powder, cellulase freeze-dried powder.
(5) each freeze-dried powder obtained above is uniformly mixed to get composite fermentation microbial inoculum.
The MRS culture medium constituent are as follows: 10 parts of peptone, 10 parts of beef extract, 5 parts of yeast extract, hydrogen citrate two
2.2 parts of ammonium, 18 parts of glucose, 1.2 parts of Tween 80,4 parts of sodium acetate, 2.5 parts of dipotassium hydrogen phosphate, 1000 parts of distilled water；Described
PCA fluid nutrient medium, constituent are as follows: 6 parts of tryptone, 2.6 parts of yeast extract, 1.8 parts of glucose, 1000 parts of distilled water.
The embedding medium is prepared from the following raw materials in parts by weight: 8 parts of maltodextrin, 8 parts of beta-cyclodextrin, and starch 12
Part, 20 parts of water.
The vacuum freeze drying parameter are as follows: 65 DEG C of condenser temperature ﹣, 30 DEG C of heating temperature, vacuum degree 60pa, drying is extremely
The moisture content of microbial inoculum is no more than 7%.
The sugarcane end pin white wine residue protein feed, preparation method includes the following steps:
(1) pretreatment of raw material: sugarcane end pin is crushed to granularity no more than 5mm, drying to moisture content is spare no more than 12%；
(2) inoculation fermentation: corn flour, dregs of beans, wheat bran, rice bran are mixed in proportion, and molasses and water is added, and are adjusted moisture and are contained
Amount is inoculated with composite fermentation microbial inoculum, barrelling is sealed by fermentation 10 days to get fermentation fine fodder to 45~60%；
(3) it is sealed by fermentation: sugarcane end pin and molasses being added in fermentation fine fodder, is pressed and sealed and continues fermentation 20 days, wherein sending out
Ferment fine fodder, sugarcane end pin, molasses three weight ratio be 1:3:1, fermentation is to there is sour slightly wine flavour to get sheath of sugarcane
Tip white wine residue protein feed.
It is described domestication bacterium acclimation method the following steps are included:
(1) it chooses bacterium mud: choosing sugarcane top stack retting mud more than half a year, it is formed above that sugar refinery bagasse laydown area stacks half a year
Sludge mixes to get original bacterium mud according to weight ratio 1:1, original bacterium mud scene is sealed up for safekeeping in sterile glass vials, experiment is taken back
It is cultivated as original flora room；
(2) primary domestication: taking fresh sugarcane end pin to be crushed to 200 mesh, in 121 DEG C of high pressure sterilization 15min, is added to after cooling
Prepared basal medium is uniformly mixed, and obtains once taming culture medium, the weight ratio of sugarcane end pin and basal medium is
1.0~1.5:1；
According to the differences of sugarcane end pin additional amount, 6 gradients are set and are respectively as follows: 1.0:1,1.1:1,1.2:1,1.3:1,1.4:1,
The different primary domestication culture medium of 6 kinds of sugarcane end pin contents is made in 1.5:1, and one group of culture medium is arranged in each gradient, and every group has
3 parallel test culture mediums；Original bacterium mud in step (1) is inoculated into each culture medium, 38 DEG C, cultivate under the conditions of pH6.4
30 days, the situation in 2~4 days one subcultures of record, culture first 12 days primary every 4 days records, and the 18th~21 day every
Primary every 3 days records, the 21st~30 day primary every 2 days records, is scored according to the case where each record.
Up to excellent with the percentage that sugarcane tail is decomposed, full marks are 10 points；It is not smelly to be excellent with bacterium solution clarification, bacterium mud,
Full marks are 10 points, and the relevant professional person of Feed Manufacturing research is artificially engaged in scoring, and 50 people of number makes even and respectively (retains two
Number) it is final score；According to above-mentioned judgment criteria, in conjunction with the situation in each group culture medium, choosing optimal one group is the
Four groups, i.e., the primary domestication bacterium tamed when the weight ratio of sugarcane end pin and basal medium is 1.3:1, qualified for domestication
Optimal primary domestication bacterium.
(3) secondary domestication: taking fresh sugarcane end pin to be crushed to 100 mesh, in 121 DEG C of high pressure sterilization 15min, obtains A material；It takes
Bagasse powder after juicing is broken to 200 mesh, in 121 DEG C of high pressure sterilization 15min, obtains B material；Take bagasse shower water shape in sugar refinery
At sewage, filter off sundries, obtain C material；
Above-mentioned A, B, C material are subjected to combination of two and three's combination respectively, each material combine be etc. weight combination, and total weight
It is identical, tetra- groups of AB, AC, BC, ABC are formed, the basal medium of material total weight a quarter is added separately to, is made multiple two
Secondary domestication culture medium, every group of culture medium for having 3 parallel tests；Sugarcane end pin and basis are cultivated in inoculation step (2) respectively
The primary domestication bacterium bacterium solution that the weight ratio of base is tamed when being 1.3:1, is cultivated 20 days under the conditions of 40 DEG C, is remembered every 2~3 days
The situation in a subculture is recorded, culture first 12 days are primary every 3 days records, and the 12nd~20 day is primary every 2 days records, according to
The case where record, scores every time.
Up to excellent with the percentage that sugarcane tail is decomposed, full marks are 10 points；It is not smelly to be excellent with bacterium solution clarification, bacterium mud,
Full marks are 10 points；The relevant professional person of Feed Manufacturing research is artificially engaged in scoring, and 50 people of number makes even and respectively (retains two
Number) it is final score；An optimal tissue culture is selected in conjunction with the situation in each group culture medium according to above-mentioned judgment criteria
Base is supported, is ABC group, bacterium solution is secondary domestication bacterium.
The basal medium is prepared from the following raw materials in parts by weight: 10 parts of peptone, 10 parts of beef extract, and yeast extract
6 parts, 8 parts of brewer's wort, 2.2 parts of diammonium hydrogen citrate, 22 parts of glucose, 0.8 part of Tween 80,5 parts of sodium acetate, dipotassium hydrogen phosphate
2.0 parts, 6 parts of tryptone, 2.8 parts of yeast extract, 2 parts of glucose, 1000 parts of distilled water.
(4) expand culture: the bacterium solution for the secondary domestication bacterium for taking step (3) to obtain expands culture, and culture medium is secondary
Culture medium when culture medium optimal situation is tamed, 45h is cultivated；
(5) it dispenses, the sub- liquid of domesticated strain can be carried out continuing to expand culture, be formed a large amount of by preservation to get the sub- liquid of domesticated strain
Dominant bacteria, to adapt to industrialized production.
Embodiment 2
A kind of sugarcane end pin white wine residue protein feed, is prepared from the following raw materials in parts by weight: 80 parts of sugarcane end pin, white wine slag 40
Part, 25 parts of molasses, 30 parts of corn flour, 10 parts of dregs of beans, 10 parts of wheat bran, 10 parts of rice bran, 0.23 part of composite fermentation microbial inoculum；
The white wine slag be using sweet potato as raw material brewing white spirit after remaining residue；
The composite fermentation microbial inoculum is prepared from the following raw materials in parts by weight: 20 parts of bacterium freeze-dried powder of domestication, candida utili
9 parts of freeze-dried powder, 7 parts of lactobacillus plantarum freeze-dried powder, 9 parts of bacillus subtilis freeze-dried powder, 5 parts of aspergillus niger freeze-dried powder, aspergillus oryzae freezes
5 parts of dry powder, 8 parts of Trichoderma viride freeze-dried powder, 19 parts of cellulase freeze-dried powder.
The composite fermentation microbial inoculum the preparation method comprises the following steps:
(1) domestication bacterium is taken to expand the bacterium solution for reaching stationary phase when culture, centrifugation takes bacterium mud to be washed with sterile phosphate buffer,
Centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out vacuum freeze drying to get domestication bacterium freeze dried powder；
(2) candida utili is inoculated in malt extract medium, culture takes bacterium to stationary phase under the conditions of 37 DEG C, pH6.4
Liquid is centrifuged, and liquid is discarded supernatant, and bacterium mud is taken to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again
Embedding medium is added in liquid, carries out vacuum freeze drying to get candida utili freeze dried powder；
(3) lactobacillus plantarum is inoculated in MRS culture medium, culture takes bacterium solution to carry out to stationary phase under the conditions of 37 DEG C, pH6.4
Centrifugation, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again, adds
Enter embedding medium, carries out vacuum freeze drying to get lactobacillus plantarum freeze dried powder；
(4) each bacterium branch is inoculated in PCA fluid nutrient medium, under the conditions of 37 DEG C, pH7.0 culture to stationary phase, take bacterium solution into
Row centrifugation, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again,
Embedding medium is added, carries out vacuum freeze drying, that is, respectively obtains bacillus subtilis freeze-dried powder, aspergillus niger freeze-dried powder, aspergillus oryzae
Freeze-dried powder, Trichoderma viride freeze-dried powder, cellulase freeze-dried powder.
(5) each freeze-dried powder obtained above is uniformly mixed to get composite fermentation microbial inoculum.
The MRS culture medium constituent are as follows: 10 parts of peptone, 9 parts of beef extract, 6 parts of yeast extract, diammonium hydrogen citrate
1.8 parts, 20 parts of glucose, 1.0 parts of Tween 80,4 parts of sodium acetate, 2.0 parts of dipotassium hydrogen phosphate, 1000 parts of distilled water；Described
PCA fluid nutrient medium, constituent are as follows: 6 parts of tryptone, 3.2 parts of yeast extract, 2 parts of glucose, 1000 parts of distilled water.
The embedding medium is prepared from the following raw materials in parts by weight: 8 parts of maltodextrin, 8 parts of beta-cyclodextrin, and starch 10
Part, 16 parts of water.
The vacuum freeze drying parameter are as follows: 65 DEG C of condenser temperature ﹣, 33 DEG C of heating temperature, vacuum degree 60pa, drying is extremely
The moisture content of microbial inoculum is no more than 7%.
The sugarcane end pin white wine residue protein feed, preparation method includes the following steps:
(1) pretreatment of raw material: sugarcane end pin is crushed to granularity no more than 5mm, drying to moisture content is spare no more than 12%；
(2) inoculation fermentation: corn flour, dregs of beans, wheat bran, rice bran are mixed in proportion, and molasses and water is added, and are adjusted moisture and are contained
Amount is inoculated with composite fermentation microbial inoculum, barrelling is sealed by fermentation 7 days to get fermentation fine fodder to 45~60%；
(3) it is sealed by fermentation: sugarcane end pin and molasses being added in fermentation fine fodder, is pressed and sealed and continues fermentation 22 days, wherein sending out
Ferment fine fodder, sugarcane end pin, molasses three weight ratio be 1:3:1, fermentation is to there is sour slightly wine flavour to get sheath of sugarcane
Tip white wine residue protein feed.
The acclimation method of the domestication bacterium is identical as embodiment 1.
Embodiment 3
A kind of sugarcane end pin white wine residue protein feed, is prepared from the following raw materials in parts by weight: 90 parts of sugarcane end pin, white wine slag 35
Part, 30 parts of molasses, 35 parts of corn flour, 13 parts of dregs of beans, 8 parts of wheat bran, 8 parts of rice bran, 0.4 part of composite fermentation microbial inoculum；
The white wine slag be using sorghum as raw material brewing white spirit after remaining residue；
The composite fermentation microbial inoculum is prepared from the following raw materials in parts by weight: 30 parts of bacterium freeze-dried powder of domestication, candida utili
8 parts of freeze-dried powder, 6 parts of lactobacillus plantarum freeze-dried powder, 8 parts of bacillus subtilis freeze-dried powder, 4 parts of aspergillus niger freeze-dried powder, aspergillus oryzae freezes
4 parts of dry powder, 7 parts of Trichoderma viride freeze-dried powder, 20 parts of cellulase freeze-dried powder.
The composite fermentation microbial inoculum the preparation method comprises the following steps:
(1) domestication bacterium is taken to expand the bacterium solution for reaching stationary phase when culture, centrifugation takes bacterium mud to be washed with sterile phosphate buffer,
Centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out vacuum freeze drying to get domestication bacterium freeze dried powder；
(2) candida utili is inoculated in malt extract medium, culture takes bacterium to stationary phase under the conditions of 37 DEG C, pH6.4
Liquid is centrifuged, and liquid is discarded supernatant, and bacterium mud is taken to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again
Embedding medium is added in liquid, carries out vacuum freeze drying to get candida utili freeze dried powder；
(3) lactobacillus plantarum is inoculated in MRS culture medium, culture takes bacterium solution to carry out to stationary phase under the conditions of 37 DEG C, pH6.4
Centrifugation, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again, adds
Enter embedding medium, carries out vacuum freeze drying to get lactobacillus plantarum freeze dried powder；
(4) each bacterium branch is inoculated in PCA fluid nutrient medium, under the conditions of 38 DEG C, pH6.8 culture to stationary phase, take bacterium solution into
Row centrifugation, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again,
Embedding medium is added, carries out vacuum freeze drying, that is, respectively obtains bacillus subtilis freeze-dried powder, aspergillus niger freeze-dried powder, aspergillus oryzae
Freeze-dried powder, Trichoderma viride freeze-dried powder, cellulase freeze-dried powder.
(5) each freeze-dried powder obtained above is uniformly mixed to get composite fermentation microbial inoculum.
The MRS culture medium constituent are as follows: 8 parts of peptone, 10 parts of beef extract, 6 parts of yeast extract, diammonium hydrogen citrate
1.5 parts, 22 parts of glucose, 0.8 part of Tween 80,5 parts of sodium acetate, 1.5 parts of dipotassium hydrogen phosphate, 950 parts of distilled water；The PCA
Fluid nutrient medium, constituent are as follows: 4 parts of tryptone, 2 parts of yeast extract, 2 parts of glucose, 950 parts of distilled water.
The embedding medium is prepared from the following raw materials in parts by weight: 9 parts of maltodextrin, 8 parts of beta-cyclodextrin, and starch 11
Part, 12 parts of water.
The vacuum freeze drying parameter are as follows: 65 DEG C of condenser temperature ﹣, 35 DEG C of heating temperature, vacuum degree 55pa, drying is extremely
The moisture content of microbial inoculum is no more than 7%.
The sugarcane end pin white wine residue protein feed, preparation method includes the following steps:
(1) pretreatment of raw material: sugarcane end pin is crushed to granularity no more than 5mm, drying to moisture content is spare no more than 12%；
(2) inoculation fermentation: corn flour, dregs of beans, wheat bran, rice bran are mixed in proportion, and molasses and water is added, and are adjusted moisture and are contained
Amount is inoculated with composite fermentation microbial inoculum, barrelling is sealed by fermentation 10 days to get fermentation fine fodder to 45~60%；
(3) it is sealed by fermentation: sugarcane end pin and molasses being added in fermentation fine fodder, is pressed and sealed and continues fermentation 20 days, wherein sending out
Ferment fine fodder, sugarcane end pin, molasses three weight ratio be 1:3:1, fermentation is to there is sour slightly wine flavour to get sheath of sugarcane
Tip white wine residue protein feed.
The acclimation method of the domestication bacterium is identical as embodiment 1.
Nutritional ingredient detection is carried out to protein feed obtained in Examples 1 to 3, result is as follows:
| Embodiment 1 | Embodiment 2 | Embodiment 3 | |
| Protein (%) | 20.08 | 20.07 | 20.15 |
| Viable count (CFU/g × 1010) | 0.377 | 0.385 | 0.398 |
| Moisture (%) | 65.36 | 65.37 | 65.42 |
| pH | 3.72 | 3.70 | 3.61 |
| Neutral detergent fiber (%) | 34.69 | 34.75 | 34.46 |
| Acidic cleaning lignin (%) | 3.45 | 3.46 | 3.38 |
| Crude fat (%) | 10.08 | 10.05 | 10.03 |
| Coarse ash (%) | 10.56 | 10.48 | 10.41 |
Application Example 1
Sugarcane end pin white wine residue protein feed obtained in the embodiment of the present invention is subjected to feeding trial to beef cattle, as follows:
It selects with a collection of beef cattle 200 similar in contemporaneity cultivation, physical condition, the kind of beef cattle is Xia Luolainiu, examination
Testing the age at initial stage is 4 months.Xia Luolainiu is equally divided into four groups, every group 50, first three groups feed Examples 1 to 3 respectively
Protein feed, control group feed the common ensilage containing bagasse, and every group is to be freely eaten, and experimental period 100 days.Test
As a result as shown in the table:
| Initial counterpoise (kg) | End of term counterpoise (kg) | Average daily gain (kg) | Feed consumption weight gain ratio | |
| Embodiment 1 | 212 | 347 | 1.35 | 6.6:1 |
| Embodiment 2 | 210 | 344 | 1.34 | 6.6:1 |
| Embodiment 3 | 216 | 353 | 1.37 | 6.7:1 |
| Control group | 215 | 290 | 0.75 | 10.1:1 |
The above results show that the beef cattle of sugarcane end pin white wine residue protein forage feed produced by the present invention has compared with ensilage
There is good gaining effect, can guarantee the healthy growth of beef cattle, the disease incidence of beef cattle is substantially reduced, and feed consumption weight gain is than reducing.
Application Example 2
Sugarcane end pin white wine residue protein feed obtained in the embodiment of the present invention is subjected to feeding trial to mutton sheep, as follows:
It selects with a collection of mutton sheep 200 similar in contemporaneity cultivation, physical condition, the kind of mutton sheep is black goat, test
Age at initial stage is 6 months.Black goat is equally divided into four groups, every group 50, first three groups feed the sugarcane of Examples 1 to 3 respectively
End pin white wine residue protein feed, control group feed the common ensilage containing sugarcane end pin, and every group is to be freely eaten, test
Phase 100 days.Test result is as shown in the table:
| Initial counterpoise (kg) | End of term counterpoise (kg) | Average daily gain (kg) | Feed consumption weight gain ratio | |
| Embodiment 1 | 25.8 | 40.5 | 0.147 | 6.7:1 |
| Embodiment 2 | 25.7 | 40.1 | 0.144 | 6.6:1 |
| Embodiment 3 | 25.7 | 40.0 | 0.143 | 6.6:1 |
| Control group | 25.7 | 32.4 | 0.067 | 8.3:1 |
The above results show that the mutton sheep of sugarcane end pin white wine residue protein forage feed produced by the present invention has compared with ensilage
There is good gaining effect, can guarantee the healthy growth of mutton sheep, than reducing, mutton is high-quality for feed consumption weight gain.
Claims (8)
1. a kind of sugarcane end pin white wine residue protein feed, it is characterised in that: be prepared from the following raw materials in parts by weight: sugarcane end pin
70~90 parts, 35~45 parts of white wine slag, 20~30 parts of molasses, 25~35 parts of corn flour, 8~13 parts of dregs of beans, 8~13 parts of wheat bran,
8~13 parts of rice bran, 0.2~0.4 part of composite fermentation microbial inoculum；
The white wine slag be using cereal or potato grain as raw material brewing white spirit after remaining residue；
The composite fermentation microbial inoculum is prepared from the following raw materials in parts by weight: 15~30 parts of bacterium freeze-dried powder of domestication, Candida utilis
8~10 parts of yeast freeze-dried powder, 6~8 parts of lactobacillus plantarum freeze-dried powder, 8~10 parts of bacillus subtilis freeze-dried powder, aspergillus niger freeze-drying
4~6 parts of powder, 4~6 parts of aspergillus oryzae freeze-dried powder, 7~10 parts of Trichoderma viride freeze-dried powder, 18~20 parts of cellulase freeze-dried powder.
2. sugarcane end pin white wine residue protein feed according to claim 1, it is characterised in that: the composite fermentation microbial inoculum
The preparation method comprises the following steps:
(1) domestication bacterium is taken to expand the bacterium solution for reaching stationary phase when culture, centrifugation takes bacterium mud to be washed with sterile phosphate buffer,
Centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out vacuum freeze drying to get domestication bacterium freeze dried powder；
(2) candida utili is inoculated in malt extract medium, is cultivated under the conditions of 36~38 DEG C, pH6.3~6.5 to steady
Periodically, it takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, centrifugation is resuspended in sterile again
Embedding medium is added in phosphate buffer, carries out vacuum freeze drying to get candida utili freeze dried powder；
(3) lactobacillus plantarum is inoculated in MRS culture medium, culture takes to stationary phase under the conditions of 36~38 DEG C, pH6.3~6.5
Bacterium solution is centrifuged, and liquid is discarded supernatant, and bacterium mud is taken to be washed with sterile phosphate buffer, and it is slow that centrifugation is resuspended in sterile phosphate again
Embedding medium is added in fliud flushing, carries out vacuum freeze drying to get lactobacillus plantarum freeze dried powder；
(4) each bacterium branch is inoculated in PCA fluid nutrient medium, is cultivated under the conditions of 36~38 DEG C, pH6.8~7.2 to stationary phase,
It takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, centrifugation is resuspended in sterile phosphate again
Buffer, be added embedding medium, carry out vacuum freeze drying, that is, respectively obtain bacillus subtilis freeze-dried powder, aspergillus niger freeze-dried powder,
Aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulase freeze-dried powder；
(5) each freeze-dried powder obtained above is uniformly mixed to get composite fermentation microbial inoculum.
3. sugarcane end pin white wine residue protein feed according to claim 1, it is characterised in that: preparation method includes following
Step:
(1) pretreatment of raw material: sugarcane end pin is crushed to granularity no more than 5mm, drying to moisture content is spare no more than 12%；
(2) inoculation fermentation: corn flour, dregs of beans, wheat bran, rice bran are mixed in proportion, and molasses and water is added, and are adjusted moisture and are contained
Amount is inoculated with composite fermentation microbial inoculum, barrelling is sealed by fermentation 5~10 days to get fermentation fine fodder to 45~60%；
(3) it is sealed by fermentation: sugarcane end pin and molasses being added in fermentation fine fodder, is pressed and sealed and continues fermentation 20~24 days,
Middle fermentation fine fodder, sugarcane end pin, molasses three weight ratio be 1:3:1, fermentation is to there is sour slightly wine flavour to get sweet
Sugarcane end pin white wine residue protein feed.
4. sugarcane end pin white wine residue protein feed according to claim 1, it is characterised in that: the domestication side of the domestication bacterium
Method the following steps are included:
(1) it chooses bacterium mud: choosing the half a year above sugarcane field soil or sugarcane top stack retting mud or sugar refinery bagasse laydown area is stacked
Half a year sludge formed above seals at original bacterium mud scene in sterile glass vials up for safekeeping to get original bacterium mud, takes back laboratory work
It is cultivated for original flora；
(2) primary domestication: taking fresh sugarcane end pin to be crushed to 100~200 mesh, cold in 121 DEG C of high pressure sterilization 10~20min
But it is added to prepared basal medium afterwards to be uniformly mixed, the weight ratio of sugarcane end pin and basal medium is 1.0~1.5:
1 to get primary domestication culture medium；
According to the difference of sugarcane end pin additional amount, 5~6 gradients are set, prepare different primary tame and docile of several sugarcane end pin contents
Change culture medium, the original bacterium mud in step (1) is inoculated into each culture medium, 35~40 DEG C, cultivate under the conditions of pH6.3~6.5
20~30 days, the situation in 2~4 days one subcultures of record was decomposed up to excellent with sugarcane tail, clear with bacterium solution
Clearly, it is excellent, in comprehensive each culture medium situation that bacterium mud is not smelly, selects one group of optimal culture medium, survives in bacterium solution
Bacterium is primary domestication bacterium；
(3) secondary domestication: taking fresh sugarcane end pin to be crushed to 100~200 mesh, in 121 DEG C of 10~20min of high pressure sterilization, obtains A
Material；It takes the bagasse powder after juicing to be broken to 100~200 mesh, in 121 DEG C of 10~20min of high pressure sterilization, obtains B material；It takes in sugar refinery
The sewage that bagasse shower water is formed filters off sundries, obtains C material；
Above-mentioned A, B, C material are subjected to combination of two or three's combination respectively, are added to basal medium, is made multiple secondary tame and docile
Change culture medium, once tames the bacterium solution of bacterium in inoculation step (2) respectively, cultivated 15~30 days under the conditions of 35~42 DEG C, every 2
Situation in~4 days one subcultures of record, decomposes up to excellent, with clarifying contaminated liquids, not smelly with sugarcane tail, bagasse
It is excellent, in comprehensive each culture medium situation, selects one group of optimal culture medium, bacterium solution is secondary domestication bacterium；
(4) expand culture: the bacterium solution for the secondary domestication bacterium for taking step (3) to obtain expands culture, and culture medium is secondary domestication
Culture medium when culture medium optimal situation cultivates 40~50h；
(5) it dispenses, the sub- liquid of domesticated strain can be carried out continuing to expand culture, be formed a large amount of by preservation to get the sub- liquid of domesticated strain
Dominant bacteria, to adapt to industrialized production.
5. sugarcane end pin white wine residue protein feed according to claim 2, it is characterised in that: the embedding medium is by following
The raw material of parts by weight is made: 8~10 parts of maltodextrin, 6~8 parts of beta-cyclodextrin, 10~12 parts of starch, 8~20 parts of water.
6. sugarcane end pin white wine residue protein feed according to claim 2, it is characterised in that: the vacuum freeze drying
Parameter are as follows: 65 DEG C of condenser temperature ﹣, 30~35 DEG C of heating temperature, vacuum degree≤60pa, the dry moisture content to microbial inoculum is no more than
7%。
7. sugarcane end pin white wine residue protein feed according to claim 2, it is characterised in that: the MRS culture medium group
At ingredient are as follows: 8~10 parts of peptone, 8~10 parts of beef extract, 4~6 parts of yeast extract, 1.5~2.2 parts of diammonium hydrogen citrate, grape
18~22 parts of sugar, 0.8~1.2 part of Tween 80,4~6 parts of sodium acetate, 1.5~2.5 parts of dipotassium hydrogen phosphate, distilled water 900~
1000 parts；The PCA fluid nutrient medium, constituent are as follows: 4~6 parts of tryptone, 2~3.2 parts of yeast extract, glucose 1~
2 parts, 950~1000 parts of distilled water.
8. sugarcane end pin white wine residue protein feed according to claim 4, it is characterised in that: the basal medium by
The raw material of following parts by weight is made: 8~10 parts of peptone, 8~10 parts of beef extract, and 4~6 parts of yeast extract, brewer's wort 5~10
Part, 1.5~2.2 parts of diammonium hydrogen citrate, 18~22 parts of glucose, 0.8~1.2 part of Tween 80,4~6 parts of sodium acetate, phosphoric acid
1.5~2.5 parts of hydrogen dipotassium, 4~6 parts of tryptone, 2~3.2 parts of yeast extract, 1~2 part of glucose, distilled water 900~1000
Part.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111186917A (en) * | 2020-01-15 | 2020-05-22 | 泸州品创科技有限公司 | Method for treating residues of brewing wastewater by using microbial agent |
| CN113273641A (en) * | 2021-05-25 | 2021-08-20 | 云南柒捌玖农业发展有限公司 | Method for producing feed by fermenting sugarcane juice |
| CN114751795A (en) * | 2022-05-11 | 2022-07-15 | 广西大学 | Method for preparing sugarcane solid fertilizer by filtering residues from culture tail water |
| CN115161227A (en) * | 2022-06-07 | 2022-10-11 | 武汉轻工大学 | Preparation method and application of probiotic culture medium |
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| CN115161227A (en) * | 2022-06-07 | 2022-10-11 | 武汉轻工大学 | Preparation method and application of probiotic culture medium |
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