CN104872379B - A kind of fermentate and its fermentation process adding the molten slurry of fish in corn-soybean meal diet - Google Patents
A kind of fermentate and its fermentation process adding the molten slurry of fish in corn-soybean meal diet Download PDFInfo
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Abstract
The present invention provides a kind of in corn bean pulp type daily ration adds the fermentate and its fermentation process of the molten slurry of fish, and each component additive amount is respectively by every gram of corn bean pulp type daily ration：Bradley yeast 2 × 109 A/g；Solve starch bud bacillus HFJ 74 × 109A/g；Lactobacillus plantarum 5 × 109A/g；Alpha amylase 20U/g；10U/ grams of cellulase；NSP complex enzymes 10U/g；Protease 100U/g；The molten slurry of fish is added, material-water ratio is adjusted, is sealed by fermentation.Present invention process is simple, it is of low cost, final products low temperature drying, vomitoxin and mycotoxin are not detected after fermented, and antigen protein significantly reduces, while being rich in a large amount of probiotics and its metabolite, the growth and breeding of the harmful bacterias such as enteropathogenic E. Coli, salmonella can effectively be inhibited, animal immunizing power is improved, and feed has strong fishlike smell after adding the molten slurry of fish, has good attractant to animal.
Description
Technical field
The invention belongs to livestock feed technical fields, and in particular to one kind adding the molten slurry of fish in corn-soybean meal diet
Fermentate and its fermentation process.
Background technology
China livestock and poultry are based on corn-soybean meal feed, and wherein corn is the main energy source of livestock and poultry, and dregs of beans is poultry
The main source of protein in fowl daily ration.
Corn can be worth highest in all cereal, be the main energy source of animal, the digestibility of starch is very high, but fine
Dimension element and non-starch polysaccharide are to influence the principal element of corn digestibility.
Dregs of beans is that most Vegetable protein sources are applied in Feed Manufacturing.Not due to soybean varieties and processing conditions
Together, nutrient and anti-nutritional factors content have larger difference.Anti-nutritional factors mainly has protease inhibitors, plant solidifying in dregs of beans
Factors, the wherein Soybean antigen proteins and oligosaccharides such as collection element, tannin, Soybean antigen protein and bad oligosaccharides have good thermostabilization
Property, they can cause the ill symptoms such as diarrhea, flatulence, be major influence factors of the dregs of beans in feed applications.
Fish meal has essential amino acid and aliphatic acid high, and carbohydrate content is low, and palatability is good, anti-nutritional factors it is few with
And can the features such as good absorption, be there is by livestock and poultry animal.But the eutrophication and intensive culture fishery of water body now
Development, fish manually mixed feed application also increasingly extensively, the demand of fish meal is huge；In addition the continuous deterioration of marine environment
With the unordered fishing of the mankind, the yield of fish meal is caused drastically to decline.The molten slurry albumen of fish is that have fresh fishes in the mistake for making fish meal
The liquid of the molten slurry of fish squeezed out in journey, it is concentrated, enzymolysis after drying form.
Invention content
The present invention provides a kind of in corn-soybean meal diet adds the fermentate and its fermentation process of the molten slurry of fish,
It ferments after the molten slurry of fish is added in main livestock and poultry diet corn-soybean meal type daily ration now, utilizes this method fermented maize-
Soybean meal based diets have fishy odor, the palatability of feed can be turned up, and are rich in a large amount of probiotics and its metabolite, inhibit to have
Evil bacterium growth, increases the immunity of animal, and high small peptide content can effectively improve the utilization rate of feed.
For achieving the above object, the present invention is achieved using following technical proposals：
The present invention provides the fermentation process that the molten slurry of fish is added in corn-soybean meal basal diet, it includes following step
Suddenly：
(1) following components is added in corn-soybean meal basal diet：Bradley yeast content is not less than 2 × 109A/g；
Bacillus amyloliquefaciens HFJ-7 contents are not less than 4 × 109A/g；Lactobacillus plantarum content is not less than 5 × 109A/g；Alphalise starch
Enzyme is not less than 20U/g；Cellulose enzyme is not less than 10U/g；NSP complex enzymes are not less than 10U/g；Protease additive amount is not less than
100U/g；It is that 20%-50% adds the molten slurry of fish according to the mass ratio of the basal diet gross mass is accounted for；
(2) plus water adjusts and expects that water quality ratio is 1:0.7-1:1, control temperature is in 30 DEG C of -50 DEG C of sealing and fermentings；
(3) fermentation is completed using 60 DEG C of drying of low temperature.
The bacillus amyloliquefaciens HFJ-7 is bacillus amyloliquefaciensBacillus amyloliquefaciens,
Deposit number is CGMCC No. 10011.
The preparation process of bacillus amyloliquefaciens HFJ-7 bacterium powders containing the bacillus amyloliquefaciens HFJ-7 is as follows：
（1）Bacillus amyloliquefaciens HFJ-7 is inoculated into the sterilizing seeding tank equipped with seed culture medium, 37 DEG C of cultures
10h, forms seed culture fluid, and the seed culture medium component includes：Molasses, 24.5 ~ 25.0 g/L；Peptone, 13.5 ~ 13.7
g/L；Potassium dihydrogen phosphate, 1.7 ~ 1.8 g/L；Sodium citrate, 1.5 ~ 1.6 g/L；Ammonium nitrate, 1.5 ~ 1.6 g/L；Magnesium sulfate, 0.5
~0.52 g/L；Adjust starting pH to 7.2；
（2）The seed culture fluid is forwarded to the production tank equipped with fermentation medium and carries out fermented and cultured, 37 DEG C of cultures
For 24 hours, when spore rate is more than 95%, stop tank, obtain zymotic fluid, the fermentation medium composition includes：Bean cake powder, 25.0 ~ 25.5
g/L；Dried Corn Steep Liquor Powder, 18.5 ~ 19.4 g/L；Sodium chloride, 4.0 ~ 4.2 g/L；Sodium dihydrogen phosphate, 2.8 ~ 2.9 g/L；Phosphoric acid hydrogen
Dipotassium, 1.2 ~ 1.3 g/L；Magnesium chloride, 0.55 ~ 0.58 g/L；Manganese sulfate, 0.15 ~ 0.16 g/L；Neutral proteinase, 2.0 ~ 2.1
g/L；Carbohydrase, 1.5 ~ 1.6 g/L；Antifoaming agent, 1.3 ~ 1.4 g/L；Adjust starting pH to 7.4, the solution starch gemma bar
The bacterium amount of bacterium HFJ-7 zymotic fluids reaches 450 × 108 CFU/g~500×108 CFU/g；
（3）It is spray-dried after the zymotic fluid is added carrier, obtains bacteria containing amount 2000 × 108 ~2200×108
The bacillus amyloliquefaciens HFJ-7 bacterium powders of CFU/g, the carrier are precipitated calcium carbonate, and the amount for adding carrier accounts for the matter of zymotic fluid
Amount ratio is：10%-15%.
Above-mentioned technical proposal is further improved：The quality of corn and dregs of beans in the corn-soybean meal basal diet
Than being 4:1-3:2.
Above-mentioned technical proposal is further improved：The step（1）The protease of middle addition is by enzyme additive amount alkalinity
Protease：Neutral proteinase：Acid protease=2:5:The mixture of 3 three kinds of components.
Above-mentioned technical proposal is further improved：The step（2）Middle sealing and fermenting 48h, aerobic hair in sealing and fermenting
Ferment 12h, anaerobic fermentation 36h.
Above-mentioned technical proposal is further improved：The step（3）The inlet air temperature of middle spray drying is 145 DEG C -150
DEG C, leaving air temp is 65 DEG C -70 DEG C.
The present invention also provides fermentates made from the fermentation process.The pH3-5 of the fermentate, lactic acid content
10-20%, small peptide content 20-30%, viable count are up to 9 × 1010cfu/g~3.8×1012cfu/g。
Compared with prior art, the present invention have effects that it is following innovation and it is effective；
1, the low temperature drying at 60 DEG C of the fermentation process in the present invention can retain a large amount of beneficial viable bacterias, while inhibit to have
Evil bacterium growth, increases the health of animal intestinal tract, improves animal immunizing power；
2, heavy albumen can be degraded to the small peptide easily absorbed in fermentation process, and content is up to 20%, be easy to animal suction
It receives, efficiency of feed utilization can be significantly improved.
3, lactic acid content is abundant in fermentate produced by the present invention, and content is up to 10%-15%, improves animal to feed
Feed intake.
4, the molten slurry of fish of present invention addition low cost ferments, and can effectively improve the smell of feed, improves adopting for animal
Appetite, while the dosage of fish meal in feed can also be reduced.
Description of the drawings
Fig. 1 is the production technological process of bacillus amyloliquefaciens HFJ-7 bacterium powders；
Fig. 2 is the high molecular weight protein experimental result of fermentation and non-fermented maize-soybean meal based diets；
Fig. 3 is the bad oligosaccharides experimental result of fermentation and non-fermented maize-soybean meal based diets；
Fig. 4 is the Escherichia coli bacteriostatic experiment result of fermentation and non-fermented maize-soybean meal based diets.
Specific implementation mode
Invention is further described in detail for present invention combination following specific examples.
Embodiment 1
One, the separation screening of HFJ-7
In January, 2012 in March, 2012, from 10, the town suitable for reading in Shouguang City of Weifang City of Shandong Province, capsicum green house of vegetables takes 30
Capsicum Rhizosphere sampling sheet.Soil sample is fully ground respectively, in heating 20min in 75 DEG C of water-baths, then uses sterile saline
In 1000 times of dilutions, in dilution spread on nutrient agar NA tablets, the scribing line purifying of picking single bacterium colony filters out 68 plants of pure bacterium altogether,
It is bacillus through microscopy, including bacterial strain HFJ-7.
The method to be stood facing each other using tablet, with Rhizoctonia solani Kuhn, Fusarium oxysporum, Pathogens Causing Root Rot Disease, cucumber fusarium axysporum disease
The soil-borne diseases disease fungus such as opportunistic pathogen, Phytophthora capsici is target disease fungus, and disease fungus is short of money between investigating 68 bacillus
The difference of anti-characteristic.The pathogen fungus block that 6mm diameters are first obtained with card punch is connected to PDA plate center, in plate surrounding, hangs down
Directly go out according to center about 3cm, is inoculated with target bacterium bacterium line, length is about 1cm.In view of the Different Kinds of Pathogens fungi speed of growth is different, stand
Withered silk kernel fungus preculture 8h, Fusarium oxysporum, Pathogens Causing Root Rot Disease, cucumber fusarium axysporum pathogen, Phytophthora capsici pathogen are trained in advance
It supports for 24 hours, then per 4 plants of bacterial strains to be tried of plating, after 28 DEG C of culture 4d, investigates bacillus amyloliquefaciens HFJ-7 and other buds
Spore bacillus is compared, to the antagonistic properties of a variety of soil-borne disease disease fungus.Record antibacterial bandwidth, i.e. two kinds of bacterium Parallel Growths it
Between aseptic area width.According to the width of antagonism band, the bacterial strain for having stronger antagonistic ability to disease fungus is chosen.
The selection result shows, 15 plants of bacterium Rhizoctonia solanis have a stronger antagonism, 13 plants of bacterium to Fusarium oxysporum have compared with
Strong antagonism, 10 plants of bacterium have stronger antagonism, 9 plants of bacterium to have stronger antagonism to make cucumber fusarium axysporum pathogen root-rot bacterium
With 7 plants of bacterium have stronger antagonism to Phytophthora capsici pathogen.Only 4 plants of bacterium simultaneously all have 5 kinds of disease fungus target bacterium
Antagonism, respectively：Bacillus amyloliquefaciens HFJ-7, bacillus subtilis G4 and G11, bacillus licheniformis D2.Wherein,
Bacillus amyloliquefaciens HFJ-7 compared with the bacillus such as withered grass, lichens, side spore, can Rhizoctonia solani, Fusarium oxysporum,
A variety of soil-borne disease disease fungus such as Pathogens Causing Root Rot Disease, cucumber fusarium axysporum pathogen, Phytophthora capsici pathogen generate apparent
Antibacterial band, is all higher than equal to 5mm, wherein most strong to the resistance of cucumber fusarium axysporum pathogen, antibacterial bandwidth is up to 8mm.
Through Bacterial Physiological biochemical method Preliminary Identification, bacterial strain HFJ-7 is bacillus, biological characteristics：Gram
Stained positive, the colony diameter formed on nutrient agar panel about 3-3.5mm, edge is irregular, intermediate fold, faint yellow.
Thalli morphology is in rod-shaped, and gemma ellipse, end is given birth to, 0.84 × 0.52 μm.
Two, the molecular genetics taxonomic identification of HFJ-7
The DNA of extraction bacterial strain HFJ-7 first, using the DNA as template, 16S rRNA universal primers are primer, to 16S
RRNA segments are expanded, and amplified fragments carry out sequencing.Sequencing result Blast softwares category kind related to GenBank
16S rRNA sequences be compared, as a result show bacillus in bacterial strain 16S rRNA sequences and GenBank gene pools
In Bacillus pumilus 16s rRNA sequence homology degree highests, homology reaches 99%.Pass through DNAMAN6.0 pairs
The 16S rRNA sequences of existing bacillus carry out phylogenetic analysis in Genbank, the results show that bacterial strain S3 16S
RRNA withBacillus amyloliquefaciensHomology highest judges the bacterial strain for the solution starch gemma of bacillus
Bacillus.
The bacillus amyloliquefaciens HFJ-7 screened is subjected to culture presevation, depositary institution：Chinese microorganism strain preservation
Administration committee's common micro-organisms center（CGMCC）；Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences
Institute of microbiology；Preservation date：On November 19th, 2014；Bacillus amyloliquefaciensBacillus amyloliquefaciensDeposit number be CGMCC No. 10011.
Three, the preparation of bacillus amyloliquefaciens bacterium powder
Bacillus amyloliquefaciens bacterium powder of the present invention is by the bacterial strain of autonomous separation screening, bacillus amyloliquefaciens HFJ-
It sprays after 7 fermentations, preparation process is as shown in Figure 1, be as follows：
（1）Seed tank culture
The bacillus amyloliquefaciens HFJ-7 bacterial strains carry out scribing line culture with NA culture mediums, and 37 DEG C of cultures for 24 hours, ensure
Spore rate>99%, as first order seed.By first order seed, it is inoculated into the sterilizing seeding tank equipped with seed culture medium, 37 DEG C
10h is cultivated, a large amount of trophosomes are formed, as secondary seed culture solution；After activation is the seed of 100% spore rate, it is inoculated into and is equipped with
In the sterilizing seeding tank of seed culture medium, 37 DEG C of culture 10h form a large amount of trophosomes, as seed culture fluid.Seed culture medium
Composition（g/L）：Molasses, 24.5；Peptone, 13.5；Potassium dihydrogen phosphate, 1.7；Sodium citrate, 1.5；Ammonium nitrate, 1.5；Sulfuric acid
Magnesium, 0.5；Added with 50% sodium hydroxide stream, adjusts starting pH to 7.2；Seed tank volume is 500L, and culture medium is settled to 300L,
Disappear condition in fact：121 DEG C of holding 30min of constant temperature；Condition of culture：37 DEG C of constant temperature+1 DEG C, tank presses 0.05Mpa, rotating speed 0-6h
130rpm、6-10h 160rpm；5.1 m of ventilation quantity 0-6h3/h、6-10h 8.8 m3/h。
（2）Fermentation tank culture
Seed culture fluid in seeding tank is forwarded to 30 tons equipped with fermentation medium production tanks and carries out fermented and cultureds, 37 DEG C
Culture for 24 hours, when spore rate is more than 95%, stops tank, obtains zymotic fluid.Fermentation medium forms（g/L）：Bean cake powder, 25.0；It is beautiful
Rice ＆ peanut milk dry powder, 18.5；Sodium chloride, 4.0；Sodium dihydrogen phosphate, 2.8；Dipotassium hydrogen phosphate, 1.2；Magnesium chloride, 0.55；Manganese sulfate,
0.15；Neutral proteinase, 2.0；Carbohydrase, 1.5；Antifoaming agent, 1.3；Added with 50% sodium hydroxide stream, adjust starting pH to
7.4；Seed tank volume is 30000L, and culture medium is settled to 21000L, and disappear condition in fact：116 DEG C of holding 30min of constant temperature；Cultivate item
Part：37 DEG C of constant temperature+1 DEG C, tank presses 0.05Mpa, rotating speed 0-10h 140rpm, 10-18h 180rpm, 18-24h 160rpm；Ventilation
Measure 0-10h 165m3/h、10-18h 215m3/h、18-24h 180m3/h.The bacillus amyloliquefaciens HFJ-7 zymotic fluids exist
Bacterium amount when putting tank commonly reaches 450 × 108 CFU/g~500×108 CFU/g。
（3）Zymotic fluid is mixed with carrier and is post-processed, bacillus amyloliquefaciens bacterium bacterium powder is obtained.The solution starch bud
Spore bacillus HFJ-7 last handling processes：Spray drying tower, LPG-6000 types enter the wind 145 DEG C -150 DEG C, and 65 DEG C -70 DEG C of outlet air carries
Body is precipitated calcium carbonate, and the mass ratio that addition carrier accounts for zymotic fluid is：10%-15%.The bacillus amyloliquefaciens HFJ-7 sieves
Point and quality check process：80 mesh sieve is crossed more than 90% bacterium powder, and with reference to withered in national standard GBT 26428-2010 Application of Direct-fed Microbials
The detection of careless bacillus measures bacillus amyloliquefaciens bacterium amount, and quality inspection is qualified, is prepared as bacillus amyloliquefaciens bacterium powder.Institute
State bacteria containing amount range 2000 × 10 in bacillus amyloliquefaciens HJF-7 bacterium powders8 ~2200×108 CFU/g。
Embodiment 2
It is uniformly mixed after taking corn 40kg and dregs of beans 10kg, the two to crush.It is not low according to material quality addition Bradley yeast
In 2 × 109A/g, solution starch bud bacillus HFJ-7 are not less than 4 × 109A/g, lactobacillus plantarum are not less than 5 × 109A/g, α-
Amylase is not less than 20U/g, and cellulase is not less than 10U/g, and NSP complex enzymes are not less than 10U/g, and protease is not less than 100U/
The molten slurry of 10kg fishes is added in 35kg water and obtains mixed liquor, mixed liquor is added directly into the feed mixed and stirred by g
It mixes uniformly, 40 DEG C are sealed by fermentation 2 days.
60 DEG C of low temperature dryings, pH4.5, lactic acid content 10.5%, small peptide content 20.1%, viable count are high after fermentation for product
Up to 9 × 1010cfu/g。
The molten slurry leading indicator of fish used in the present invention（It buys in Shandong Rongcheng）：pH 5.71；Moisture 45.67%；Thick egg
White 36.78%；The molten protein 65 .99% of acid；Ash content 12.58%；Volatile Base Nitrogen（VBN） 332.64mg/100g；Amino
Sour summation：32.5%.
It can be seen that the molten slurry protein content of fish is very high by the above index, and be largely small molecular protein.Since fish is molten
Slurry is the by-product of fish meal processing, therefore has strong fishlike smell, can improve the attractant of livestock and poultry, therefore develop the molten slurry of fish
It recycles and using with objective economic benefit and social benefit.
Embodiment 3
It is uniformly mixed after taking corn 30kg and dregs of beans 20kg, the two to crush.According to material quality addition Bradley yeast 2 ×
109A/gram, solution starch bud bacillus (HFJ-7) 4 × 109A/gram, lactobacillus plantarum 5 × 109A/gram, 20U/ grams of alpha-amylase,
The molten slurry of 25kg fishes is added in 45kg water and is mixed by 10U/ grams of cellulase, NSP complex enzyme 10U/g, protease 100U/g
The mixed liquor mixed is added directly into the feed mixed and stirs evenly by liquid, and 40 DEG C are sealed by fermentation 2 days.
60 DEG C of low temperature dryings, pH3.8, lactic acid content 18.5%, small peptide content 25.3%, viable count are high after fermentation for product
Up to 3.8 × 1012cfu/g。
Embodiment 4 is fermented compare related to non-fermented maize-soybean meal based diets
1）High molecular weight protein
1. learn from else's experience fermentation and unfermentable feed 0.2g respectively, add 30ml 0.6%KOH, magnetic agitation 1h under room temperature,
4000rpm centrifuges 10min, takes supernatant；
2. 100ul supernatant+100ul loading buffer boil 8min after mixing；
3. taking 10ul, the point sample on 12% SDS-pAGE glue keeps constant current 15mA, about 40-50min in 5% concentration glue；
Into separation gel, electric current is adjusted to 20mA, until forward position is close at the 1cm of ground；
4. shelling glue poststaining 2h, decoloration is overnight；
Experimental result is shown in Fig. 2, as can be seen from Figure 2：Antigen protein is the principal element for causing grice diarrhoea in dregs of beans,
Fermentation group antigen protein is substantially reduced than group of not fermenting.
2）Bad oligosaccharides
1. taking fermentation group respectively and being fermentation group 10g samples, add 80% ethyl alcohol of 50ml, 70 DEG C of extraction 1h are collected after filtering
Clear liquid；
2. being concentrated by evaporation in 90 DEG C of water-baths, 150ml is settled to 80% ethyl alcohol；
3. 10000rpm centrifuges 10min, takes supernatant, measured for oligosaccharides；
4. TLC tablets activate 30min in 105 DEG C, 5ul point samples are taken after cooling, use solvent（Acetonitrile：Glacial acetic acid：Water=6:
3:1）Expansion, after to be deployed dose drowned, takes out TLC tablets, and unfolded surface agent is dried up with hair-dryer；
5. in 105 DEG C of drying in oven 30min, with the organic solvent that disperses, developing solution（+ 0.5% 3,5-dihydroxy of 20% sulfuric acid
Base toluene）Middle immersion 15s, hair-dryer cold wind dry up surface, and then develop the color in 105 DEG C 3-5min, until showing clear sugared point.
Test result as shown in Figure 3, as can be seen from Figure 3：Bad oligosaccharides is the principal element for causing piglet flatulence, hair
Ferment group raffinose and stachyose significantly reduce.
3）Escherichia coli bacteriostatic test
1. 100ul Escherichia coli bacteria liquids is taken uniformly to be coated on above nutrient agar；
2. taking fermentation group and group sample 20g that do not ferment, 20ml sterile waters are added and stir evenly, stand 15min；
3. taking the supernatant 100ul stood respectively, it is added in the Oxford cup for being placed on E. coli plate；
4. 37 DEG C of cultures are for 24 hours.
Test result as shown in Figure 4, as can be seen from Figure 4：Fermentation group is by Bradley yeast, bacillus amyloliquefaciens
A large amount of beneficial products can be generated after fermenting with lactobacillus plantarum, apparent inhibition zone is generated to Escherichia coli, can effectively inhibit big
The growth of enterobacteria has certain curative effect to treatment grice diarrhoea.
Embodiment 5, culture experiment
1）Diarrhea rate is tested
Piglet is divided into two groups, every group 30 according to weight immediately.Experimental group is fermented maize-soybean meal based diets, control
Group is non-fermented maize-soybean meal based diets, and two groups of piglets are fed 30 days, records related data
Experimental result is as shown in Table 1 and Table 2：
1 diarrhea rate experimental result of table
| Group | Experimental group | Control group |
| Piglet sum（Head） | 30 | 30 |
| Diarrhea rate | 1.17% | 3.17% |
| The death rate | 0 | 0 |
Note：Diarrhea rate=and diarrhea pig's head number of days/(pig's head sum * feeds number of days）* 100%
2 feeding trial related data of table
| Group | Experimental group | Control group |
| Piglet sum（Head） | 30 | 30 |
| Average daily gain（G/ days/head） | 821.1 | 779.2 |
| Average daily gain（G/ days/head） | 456.17 | 397.55 |
| Feedstuff-meat ratio | 1.8 | 1.96 |
Interpretation of result：As shown in Table 1 and Table 2, piglet death, but experimental group diarrhea do not occur for experimental group and control group
Rate is significantly lower than control group.After the molten slurry fermentation of fish is added, feed attractant is preferable, and experimental group average daily gain is higher than control
Group, for the feedstuff-meat ratio of experimental group also above control group, the corn-soybean meal type daily ration after fermentation is conducive to digesting and assimilating for piglet.
The above embodiments are merely illustrative of the technical solutions of the present invention, rather than is limited；Although with reference to aforementioned reality
Applying example, invention is explained in detail, for those of ordinary skill in the art, still can be to aforementioned implementation
Technical solution recorded in example is modified or equivalent replacement of some of the technical features；And these are changed or replace
It changes, the spirit and scope for claimed technical solution of the invention that it does not separate the essence of the corresponding technical solution.
Claims (6)
1. a kind of fermentation process adding the molten slurry of fish in corn-soybean meal diet, it is characterised in that it includes the following steps：
(1) following components is added in corn-soybean meal basal diet：Bradley yeast content is not less than 2 × 109A/g；Solve starch
Bacillus HFJ-7 contents are not less than 4 × 109A/g；Lactobacillus plantarum content is not less than 5 × 109A/g；Alpha-amylase is not low
In 20U/g；Cellulase is not less than 10U/g；NSP complex enzymes are not less than 10U/g；Protease additive amount is not less than 100U/g；It presses
It is that 20%-50% adds the molten slurry of fish according to the mass ratio of the basal diet gross mass is accounted for；
(2) plus water adjusts and expects that water quality ratio is 1:0.7-1:1, control temperature is in 30 DEG C of -50 DEG C of sealing and fermentings；
(3) fermentation is completed using 60 DEG C of drying of low temperature；
The bacillus amyloliquefaciens HFJ-7 is bacillus amyloliquefaciensBacillus amyloliquefaciens, preservation
Number is CGMCC No. 10011；
The preparation process of bacillus amyloliquefaciens HFJ-7 bacterium powders containing the bacillus amyloliquefaciens HFJ-7 is as follows：
（1）Bacillus amyloliquefaciens HFJ-7 is inoculated into the sterilizing seeding tank equipped with seed culture medium, 37 DEG C of culture 10h, shape
At seed culture fluid, the seed culture medium component includes：Molasses, 24.5 ~ 25.0 g/L；Peptone, 13.5 ~ 13.7 g/L；
Potassium dihydrogen phosphate, 1.7 ~ 1.8 g/L；Sodium citrate, 1.5 ~ 1.6 g/L；Ammonium nitrate, 1.5 ~ 1.6 g/L；Magnesium sulfate, 0.5 ~
0.52 g/L；Adjust starting pH to 7.2；
（2）The seed culture fluid is forwarded to the production tank equipped with fermentation medium and carries out fermented and cultured, 37 DEG C are cultivated for 24 hours,
When spore rate is more than 95%, stop tank, obtain zymotic fluid, the fermentation medium composition includes：Bean cake powder, 25.0 ~ 25.5 g/
L；Dried Corn Steep Liquor Powder, 18.5 ~ 19.4 g/L；Sodium chloride, 4.0 ~ 4.2 g/L；Sodium dihydrogen phosphate, 2.8 ~ 2.9 g/L；Phosphoric acid hydrogen two
Potassium, 1.2 ~ 1.3 g/L；Magnesium chloride, 0.55 ~ 0.58 g/L；Manganese sulfate, 0.15 ~ 0.16 g/L；Neutral proteinase, 2.0 ~ 2.1
g/L；Carbohydrase, 1.5 ~ 1.6 g/L；Antifoaming agent, 1.3 ~ 1.4 g/L；Adjust starting pH to 7.4, the solution starch gemma bar
The bacterium amount of bacterium HFJ-7 zymotic fluids reaches 450 × 108 CFU/g~500×108CFU/g；
（3）It is spray-dried after the zymotic fluid is added carrier, obtains bacteria containing amount 2000 × 108 ~2200×108 CFU/g
Bacillus amyloliquefaciens HFJ-7 bacterium powders, the carrier is precipitated calcium carbonate, and the amount for adding carrier accounts for the mass ratio of zymotic fluid
For：10%-15%；
The step（1）The protease of middle addition is by enzyme additive amount alkali protease：Neutral proteinase：Acid protease=2:
5:The mixture of 3 three kinds of components.
2. the biofermentation method according to claim 1 for adding the molten slurry of fish in corn-soybean meal diet, feature exist
In：The mass ratio of corn and dregs of beans is 4 in the corn-soybean meal basal diet:1-3:2.
3. the biofermentation method according to claim 1 for adding the molten slurry of fish in corn-soybean meal diet, feature exist
In：The step（2）Middle sealing and fermenting 48h, aerobic fermentation 12h, anaerobic fermentation 36h in sealing and fermenting.
4. the biofermentation method according to claim 3 for adding the molten slurry of fish in corn-soybean meal diet, feature exist
In：The step（3）The inlet air temperature of middle spray drying is 145 DEG C -150 DEG C, and leaving air temp is 65 DEG C -70 DEG C.
5. fermentate made from claim 1-4 any one of them fermentation process.
6. fermentate according to claim 5, it is characterised in that：The pH3-5 of the fermentate, lactic acid content 10-20%,
Small peptide content 20-30%, viable count are up to 9 × 1010cfu/g~3.8×1012cfu/g。
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| CN105779423A (en) * | 2015-11-17 | 2016-07-20 | 济南诺能生物工程有限公司 | Compound protease as well as production method and applications thereof |
| CN110651898A (en) * | 2019-11-11 | 2020-01-07 | 白城市隆盛实业科技有限公司 | Fermentation method of soybean meal |
| CN113632878A (en) * | 2021-08-09 | 2021-11-12 | 云南师范大学 | Enzyme-bacterium composite biological agent for degrading mycotoxin in fermented unconventional feed and application |
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