CN104872379A - Fermentation substance prepared by adding fish soluble in corn-soybean meal diet and fermentation method of fermentation substance - Google Patents
Fermentation substance prepared by adding fish soluble in corn-soybean meal diet and fermentation method of fermentation substance Download PDFInfo
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Abstract
The invention provides a fermentation substance prepared by adding fish soluble in corn-soybean meal diet and a fermentation method of the fermentation substance; according to per gram of the corn-soybean meal diet, the additive amounts of all components are as follows: 2*10<9>/g of saccharomyces boulardii, 4*10<9>/g of bacillus amyloliquefaciens HFJ-7, 5*10<9>/g of lactobacillus plantarum, 20U/g of alpha-amylase, 10U/g of cellulose, 10U/g of NSP (Non Starch Polysaccharides) composite enzyme, and 100U/g of protease; fish soluble is added, the material-to-water ratio is adjusted, and the fermentation is carried out in a sealed environment. The process is simple, the cost is low, a finished product is dried at low temperature, vomitoxin and mycotoxin cannot be detected after fermentation, the content of antigen protein is reduced remarkably, the diet is rich in probiotics and metabolic products, the growth and the reproduction of pathogenic escherichia coli, paratyphoid and other harmful germs can be effectively inhibited, the animal immunity is improved, a feed has the rich fishlike smell after addition of the fish soluble, and has the good food calling property on animals.
Description
Technical field
The invention belongs to livestock feed technical field, be specifically related to a kind of in corn-soybean meal diet, add the molten slurry of fish fermentate and fermentation process thereof.
Background technology
China livestock and poultry are based on corn-soybean meal feed, and wherein corn is the main energy sources of livestock and poultry, and dregs of beans is the main source of protein in livestock and poultry diet.
Corn can be worth the highest in all cereal, and be the main energy source of animal, the digestibility of starch is very high, but cellulose and SNSP are the principal elements affecting corn digestibility.
Dregs of beans applies maximum Vegetable protein sources in Feed Manufacturing.Due to the difference of soybean varieties and processing conditions, its nutrient and ANFs content have larger difference.In dregs of beans, ANFs mainly contains the factors such as protease inhibitors, phytolectin, tannin, Soybean antigen protein and bad oligosaccharides, wherein Soybean antigen protein and oligosaccharides have good heat endurance, they can cause the ill symptomses such as diarrhoea, flatulence, are the major influence factors of dregs of beans in feed applications.
Fish meal has essential amino acid and aliphatic acid is high, and carbohydrate content is low, good palatability, and ANFs is few and can be had the features such as well absorption by livestock and poultry animal.But the eutrophication of water body and the development of intensive aquaculture industry now, the application of fish artifical compound feed is also day by day extensive, and the demand of fish meal is huge; The continuous deterioration of marine environment and the unordered of the mankind are fished in addition, cause the output of fish meal sharply to decline.Fish molten slurry albumen has fresh fishes making the liquid of the molten slurry of fish squeezed out in the process of fish meal, and through concentrating, after enzymolysis, drying forms.
Summary of the invention
The invention provides a kind of in corn-soybean meal diet, add the molten slurry of fish fermentate and fermentation process thereof, ferment add the molten slurry of fish in livestock and poultry diet corn-soybean meal type daily ration main now after, utilize the method fermented maize-soybean meal based diets, there is fishy smell taste, the palatability of feed can be heightened, be rich in a large amount of probio and metabolite thereof, suppress harmful bacteria growth, increase the immunity of animal, higher primary school's peptide content, effectively can improve the utilization rate of feed.
For achieving the above object, the present invention adopts following technical proposals to be achieved:
The invention provides the fermentation process adding the molten slurry of fish in corn-soybean meal basal diet, it comprises the following steps:
(1) in corn-soybean meal basal diet, following component is added: Bradley amount of yeast is not less than 2 × 10
9individual/g; Bacillus amyloliquefaciens HFJ-7 content is not less than 4 × 10
9individual/g; Lactobacillus plantarum content is not less than 5 × 10
9individual/g; AMS is not less than 20U/g; Cellulose enzyme is not less than 10U/g; NSP complex enzyme is not less than 10U/g; Protease addition is not less than 100U/g; Be that 20%-50% adds fish molten slurry according to the mass ratio accounting for described basal diet gross mass;
(2) add water adjustment material water quality than being 1:0.7-1:1, control temperature is 30 DEG C of-50 DEG C of sealing and fermenting;
(3) fermented and adopted low temperature 60 DEG C oven dry.
Described bacillus amyloliquefaciens HFJ-7 is bacillus amyloliquefaciens
bacillus amyloliquefaciens, its deposit number is CGMCC No. 10011.
The preparation process of the bacillus amyloliquefaciens HFJ-7 bacterium powder containing described bacillus amyloliquefaciens HFJ-7 is as follows:
(1) be inoculated into by bacillus amyloliquefaciens HFJ-7 and be equipped with in the sterilizing seeding tank of seed culture medium, cultivate 10h for 37 DEG C, form seed culture fluid, described seed culture medium component comprises: molasses, 24.5 ~ 25.0 g/L; Peptone, 13.5 ~ 13.7 g/L; Potassium dihydrogen phosphate, 1.7 ~ 1.8 g/L; Natrium citricum, 1.5 ~ 1.6 g/L; Ammonium nitrate, 1.5 ~ 1.6 g/L; Magnesium sulfate, 0.5 ~ 0.52 g/L; Regulate initial pH to 7.2;
(2) described seed culture fluid is forwarded to the production tank that fermentation medium is housed and carries out fermented and cultured, cultivate 24h, when spore rate is more than 95%, stop tank for 37 DEG C, obtain zymotic fluid, described fermentation medium composition comprises: 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; Dipotassium hydrogen phosphate, 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; Defoamer, 1.3 ~ 1.4 g/L; Regulate initial pH to 7.4, the bacterium amount of described bacillus amyloliquefaciens HFJ-7 zymotic fluid reaches 450 × 10
8cFU/g ~ 500 × 10
8cFU/g;
(3) carry out spraying dry after described zymotic fluid being added carrier, obtain bacteria containing amount 2000 × 10
8~ 2200 × 10
8the bacillus amyloliquefaciens HFJ-7 bacterium powder of CFU/g, described carrier is precipitated calcium carbonate, adds the mass ratio that the amount of carrier accounts for zymotic fluid to be: 10%-15%.
Further improvement to technique scheme: in described corn-soybean meal basal diet, the mass ratio of corn and dregs of beans is 4:1-3:2.
Further improvement to technique scheme: the protease added in described step (1) is by enzyme addition alkali protease: neutral proteinase: the mixture of three kinds of components of acid protease=2:5:3.
Further improvement to technique scheme: sealing and fermenting 48h in described step (2), aerobic fermentation 12h in sealing and fermenting, anaerobic fermentation 36h.
Further improvement to technique scheme: in described step (3), spray-dired EAT is 145 DEG C-150 DEG C, leaving air temp is 65 DEG C-70 DEG C.
Present invention also offers the fermentate that described fermentation process is obtained.The pH3-5 of described fermentate, lactic acid content 10-20%, little peptide content 20-30%, viable count is up to 9 × 10
10cfu/g ~ 3.8 × 10
12cfu/g.
The present invention compared with prior art, has following innovation and effective effect;
1, the fermentation process in the present invention is low temperature drying at 60 DEG C, can retain a large amount of useful viable bacteria, suppresses harmful bacteria growth simultaneously, increases the health of animal intestinal, improve animal immunizing power;
2, in sweat, heavy albumen can be degraded to the little peptide easily absorbed, and content, up to 20%, is easy to animal and absorbs, can significantly improve efficiency of feed utilization.
3, in the fermentate that obtains of the present invention, lactic acid content enriches, and content, up to 10%-15%, improves the feed intake of animal to feed.
4, the molten slurry of fish that the present invention adds low cost ferments, and effectively can improve the smell of feed, improves the feed intake of animal, can also reduce the consumption of fish meal in feed simultaneously.
Accompanying drawing explanation
Fig. 1 is the production technological process of bacillus amyloliquefaciens HFJ-7 bacterium powder;
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.
Detailed description of the invention
The present invention is described in further detail the present invention in conjunction with following specific embodiment.
Embodiment 1
One, the separation screening of HFJ-7
In year March in January, 2012 to 2012, from Shouguang City of Weifang City of Shandong Province, 10, town suitable for reading capsicum green house of vegetables gets 30 capsicum Rhizosphere samplings originally.Soil sample is fully grinding respectively, in 75 DEG C of water-baths, heat 20min, then with 1000 times of dilutions in SPSS, dilution spread on nutrient agar NA flat board, picking list bacterium colony line purifying, filters out the pure bacterium of 68 strain altogether, bacillus is, comprising bacterial strain HFJ-7 through microscopy.
Adopt the method for dull and stereotyped face-off, with soil-borne disease disease funguses such as Rhizoctonia solani Kuhn, Fusarium oxysporum, Pathogens Causing Root Rot Disease, cucumber fusarium axysporum pathogen, Phytophthora capsicis for target disease fungus, investigate the difference of disease fungus antagonistic properties between 68 bacillus.First obtain the pathogen bacterium block of 6mm diameter with card punch, be connected to the dull and stereotyped central authorities of PDA, in plate surrounding, be vertically about 3cm according to center and go out, inoculation target bacterium bacterium line, length is about 1cm.Consider that Different Kinds of Pathogens conk speed is different, Rhizoctonia solani Kuhn preculture 8h, Fusarium oxysporum, Pathogens Causing Root Rot Disease, cucumber fusarium axysporum pathogen, Phytophthora capsici pathogen preculture 24h, then every plating 4 strain is waited to try bacterial strain, after 28 DEG C of cultivation 4d, investigate bacillus amyloliquefaciens HFJ-7 compared with other bacillus, to the antagonistic properties of multiple soil-borne disease disease fungus.Record antibacterial bandwidth, the aseptic area width namely between two kinds of bacterium Parallel Growths.According to the width of antagonism band, choose bacterial strain disease fungus being had to stronger antagonistic ability.
The selection result shows, 15 strain bacterium Rhizoctonia solani have stronger antagonism, 13 strain bacterium have stronger antagonism to Fusarium oxysporum, 10 strain bacterium have stronger antagonism to root-rot bacterium, 9 strain bacterium have stronger antagonism to cucumber fusarium axysporum pathogen, and 7 strain bacterium have stronger antagonism to Phytophthora capsici pathogen.Only there are 4 strain bacterium all to have antagonism to 5 kinds of disease fungus target bacterium simultaneously, are respectively: bacillus amyloliquefaciens HFJ-7, bacillus subtilis G4 and G11, bacillus licheniformis D2.Wherein, bacillus amyloliquefaciens HFJ-7 is compared with the bacillus such as withered grass, lichens, side spore, can the multiple soil-borne disease disease fungus such as Rhizoctonia solani, Fusarium oxysporum, Pathogens Causing Root Rot Disease, cucumber fusarium axysporum pathogen, Phytophthora capsici pathogen, produce obvious antibacterial band, all be more than or equal to 5mm, wherein the strongest 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, its biological characteristics: Gram-positive, and the colony diameter that nutrient agar panel is formed is about 3-3.5mm, and edge is irregular, and middle fold is faint yellow.Thalli morphology is shaft-like, and gemma is oval, and end is raw, 0.84 × 0.52 μm.
Two, the molecular genetics taxonomic identification of HFJ-7
First extract the DNA of bacterial strain HFJ-7, with this DNA for template, 16S rRNA universal primer is primer, and increase to 16S rRNA fragment, amplified fragments carries out sequencing.Sequencing result Blast software the belong to 16S rRNA sequence of planting relevant to GenBank is compared, the 16s rRNA sequence homology degree that result shows the Bacillus pumilus in this bacterial strain 16S rRNA sequence and GenBank gene pool in bacillus is the highest, and homology reaches 99%.Carry out phylogenetic analysis by the 16S rRNA sequence of DNAMAN6.0 to bacillus existing in Genbank, result show, bacterial strain S3 16S rRNA with
bacillus amyloliquefacienshomology is the highest, judges the bacillus amyloliquefaciens of this bacterial strain as bacillus.
The bacillus amyloliquefaciens HFJ-7 screened is carried out culture presevation, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC); Address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica; Preservation date: on November 19th, 2014; Bacillus amyloliquefaciens
bacillus 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, and after bacillus amyloliquefaciens HFJ-7 ferments, spraying forms, and as shown in Figure 1, concrete steps are as follows for its preparation process:
(1) seed tank culture
Described bacillus amyloliquefaciens HFJ-7 bacterial strain, carries out line with NA culture medium and cultivates, and cultivates 24h for 37 DEG C, and guarantee spore rate >99%'s, as first order seed.By first order seed, be inoculated into and be equipped with in the sterilizing seeding tank of seed culture medium, cultivate 10h, form a large amount of trophosome, as secondary seed nutrient solution for 37 DEG C; Activation is after the seed of 100% spore rate, is inoculated into and is equipped with in the sterilizing seeding tank of seed culture medium, cultivates 10h, forms a large amount of trophosome, as seed culture fluid for 37 DEG C.Seed culture medium composition (g/L): molasses, 24.5; Peptone, 13.5; Potassium dihydrogen phosphate, 1.7; Natrium citricum, 1.5; Ammonium nitrate, 1.5; Magnesium sulfate, 0.5; Add with 50% NaOH stream, regulate initial pH to 7.2; Seeding tank volume is 500L, and culture medium is settled to 300L, and disappear condition in fact: constant temperature 121 DEG C keeps 30min; Condition of culture: constant temperature 37 DEG C
+1 DEG C, tank pressure 0.05Mpa, rotating speed 0-6h 130rpm, 6-10h 160rpm; Ventilation 0-6h 5.1 m
3/ h, 6-10h 8.8 m
3/ h.
(2) fermentation tank culture
Seed culture fluid in seeding tank is forwarded to 30 tons of production tanks that fermentation medium is housed and carries out fermented and cultured, cultivate 24h, when spore rate is more than 95%, stop tank for 37 DEG C, obtain zymotic fluid.Fermentation medium composition (g/L): bean cake powder, 25.0; Dried Corn Steep Liquor 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; Defoamer, 1.3; Add with 50% NaOH stream, regulate initial pH to 7.4; Seeding tank volume is 30000L, and culture medium is settled to 21000L, and disappear condition in fact: constant temperature 116 DEG C keeps 30min; Condition of culture: constant temperature 37 DEG C
+1 DEG C, tank pressure 0.05Mpa, rotating speed 0-10h 140rpm, 10-18h 180rpm, 18-24h 160rpm; Ventilation 0-10h 165m
3/ h, 10-18h 215m
3/ h, 18-24h 180m
3/ h.The bacterium amount of described bacillus amyloliquefaciens HFJ-7 zymotic fluid when putting tank generally reaches 450 × 10
8cFU/g ~ 500 × 10
8cFU/g.
(3) zymotic fluid is mixed with carrier carry out post processing, obtain bacillus amyloliquefaciens bacterium bacterium powder.Described bacillus amyloliquefaciens HFJ-7 last handling process: spray drying tower, LPG-6000 type, air intake 145 DEG C-150 DEG C, air-out 65 DEG C-70 DEG C, carrier is precipitated calcium carbonate, and the mass ratio that interpolation carrier accounts for zymotic fluid is: 10%-15%.Described bacillus amyloliquefaciens HFJ-7 sieves and quality check process: all cross 80 mesh sieves more than 90% bacterium powder, and with reference to the detection of bacillus subtilis in GB GBT 26428-2010 Application of Direct-fed Microbials, measure bacillus amyloliquefaciens bacterium amount, quality inspection is qualified, is prepared as bacillus amyloliquefaciens bacterium powder.Bacteria containing amount scope 2000 × 10 in described bacillus amyloliquefaciens HJF-7 bacterium powder
8~ 2200 × 10
8cFU/g.
Embodiment 2
Get corn 40kg and dregs of beans 10kg, both mix after pulverizing.Add Bradley yeast according to material quality and be not less than 2 × 10
9individual/g, separates starch bud bacillus HFJ-7 and is not less than 4 × 10
9individual/g, Lactobacillus plantarum is not less than 5 × 10
9individual/g, AMS is not less than 20U/g, cellulase is not less than 10U/g, NSP complex enzyme is not less than 10U/g, protease is not less than 100U/g, molten for 10kg fish slurry is added in 35kg water and obtains mixed liquor, mixed liquor is directly joined in the described feed mixed and stir, 40 DEG C of sealing and fermenting 2 days.
60 DEG C of low temperature dryings after product fermentation ends, pH4.5, lactic acid content 10.5%, little peptide content 20.1%, viable count is up to 9 × 10
10cfu/g.
The molten slurry leading indicator of fish (buying in Rongcheng, Shandong) that the present invention is used: pH 5.71; Moisture 45.67%; Crude protein 36.78%; Acid-soluble protein 65 .99%; Ash content 12.58%; VBN (VBN) 332.64mg/100g; Amino acid summation: 32.5%.
Can see that fish molten slurry protein content is very high by above index, and major part is small molecular protein.Due to the accessory substance that the molten slurry of fish is fish meal processing, therefore have strong fishlike smell, can improve the attractant of livestock and poultry, recovery and the utilization of therefore developing the molten slurry of fish have objective economic benefit and social benefit.
Embodiment 3
Get corn 30kg and dregs of beans 20kg, both mix after pulverizing.Bradley yeast 2 × 10 is added according to material quality
9individual/gram, separate starch bud bacillus (HFJ-7) 4 × 10
9individual/gram, Lactobacillus plantarum 5 × 10
9individual/gram, AMS 20U/ gram, cellulase 10U/ gram, NSP complex enzyme 10U/g, protease 100U/g, add to molten for 25kg fish slurry in 45kg water and obtain mixed liquor, the mixed liquor mixed directly is joined in the described feed mixed and stirs, 40 DEG C of sealing and fermenting 2 days.
60 DEG C of low temperature dryings after product fermentation ends, pH3.8, lactic acid content 18.5%, little peptide content 25.3%, viable count is up to 3.8 × 10
12cfu/g.
Embodiment 4, fermentation and non-fermented maize-soybean meal based diets correlation ratio are comparatively
1) high molecular weight protein
1. learn from else's experience fermentation and unfermentable feed 0.2g respectively, adds 30ml 0.6%KOH, and normal temperature lower magnetic force stirs 1h, 4000rpm, and centrifugal 10min, gets supernatant;
2. 100ul supernatant+100ul loading buffer, boils 8min after mixing;
3. get 10ul, point sample on 12% SDS-pAGE glue, in 5% concentrated glue, keep constant current 15mA, about 40-50min; Enter separation gel, regulate electric current to 20mA, until forward position is close to 1cm place, ground;
4. shell glue poststaining 2h, decolouring is spent the night;
Experimental result is shown in Fig. 2, and as can be seen from Figure 2: antigen protein is the principal element causing grice diarrhoea in dregs of beans, fermentation group antigen protein has obvious reduction than group of not fermenting.
2) bad oligosaccharides
1. get fermentation group respectively and for fermentation group 10g sample, add 50ml 80% ethanol, 70 DEG C are extracted 1h, collect clear liquid after filtration;
2. evaporation and concentration in 90 DEG C of water-baths, is settled to 150ml with 80% ethanol;
3. 10000rpm, centrifugal 10min, gets supernatant, measures for oligosaccharides;
4. TLC is dull and stereotyped gets 5ul point sample in 105 DEG C of activation 30min, launches with solvent (acetonitrile: glacial acetic acid: water=6:3:1) after cooling, to be deployed dose drowned after, take out TLC dull and stereotyped, dry up unfolded surface agent with hair-dryer;
5. in 105 DEG C of drying in oven 30min, with the organic solvent that disperses, soak 15s in nitrite ion (20% sulfuric acid+0.5% 3,5-orcin), hair-dryer cold wind dries up surface, and then develop the color 3-5min in 105 DEG C, until show clear sugared point.
Result of the test as shown in Figure 3, as can be seen from Figure 3: bad oligosaccharides is the principal element causing piglet flatulence, and fermentation group raffinose and stachyose obviously reduce.
3) Escherichia coli bacteriostatic test
1. get 100ul Escherichia coli bacteria liquid to be coated on uniformly above nutrient agar;
2. get fermentation group and the group sample 20g that do not ferment, add 20ml sterilized water and stir, leave standstill 15min;
3. get the supernatant 100ul left standstill respectively, be added to and be placed in the Oxford cup of E. coli plate;
4. 24h is cultivated for 37 DEG C.
Result of the test as shown in Figure 4, as can be seen from Figure 4: fermentation group can produce a large amount of beneficial products after the fermentation of Bradley yeast, bacillus amyloliquefaciens and Lactobacillus plantarum, obvious inhibition zone is produced to Escherichia coli, can effectively suppress colibacillary growth, have certain curative effect to treatment grice diarrhoea.
Embodiment 5, culture experiment
1) diarrhea rate experiment
Piglet is divided into two groups immediately according to body weight, often organizes 30.Experimental group is fermented maize-soybean meal based diets, and control group is non-fermented maize-soybean meal based diets, is fed 30 days by two groups of piglets, record related data
Experimental result is as shown in Table 1 and Table 2:
Table 1 diarrhea rate experimental result
| Group | Experimental group | Control group |
| Piglet sum (head) | 30 | 30 |
| Diarrhea rate | 1.17% | 3.17% |
| The death rate | 0 | 0 |
Note: diarrhea rate={ diarrhoea pig's head number of days/(pig's head sum * feed number of days) } * 100%
Table 2 feeding trial related data
| Group | Experimental group | Control group |
| Piglet sum (head) | 30 | 30 |
| Average daily ingestion amount (g/ day/head) | 821.1 | 779.2 |
| Average daily gain (g/ day/head) | 456.17 | 397.55 |
| Feedstuff-meat ratio | 1.8 | 1.96 |
Interpretation of result: as shown in Table 1 and Table 2, all there is not piglet death in experimental group and control group, but experimental group diarrhea rate is starkly lower than control group.After adding the molten slurry fermentation of fish, feed attractant is better, and the average daily ingestion amount of experimental group is higher than control group, and the feedstuff-meat ratio of experimental group is also higher than control group, and the corn-soybean meal type daily ration after fermentation is conducive to digesting and assimilating of piglet.
Above embodiment only in order to technical scheme of the present invention to be described, but not is limited; Although with reference to previous embodiment to invention has been detailed description, for the person of ordinary skill of the art, still can modify to the technical scheme described in previous embodiment, or equivalent replacement is carried out to wherein portion of techniques feature; And these amendments or replacement, do not make the essence of appropriate technical solution depart from the spirit and scope of the present invention's technical scheme required for protection.
Claims (9)
1. in corn-soybean meal diet, add a fermentation process for the molten slurry of fish, it is characterized in that it comprises the following steps:
(1) in corn-soybean meal basal diet, following component is added: Bradley amount of yeast is not less than 2 × 10
9individual/g; Bacillus amyloliquefaciens HFJ-7 content is not less than 4 × 10
9individual/g; Lactobacillus plantarum content is not less than 5 × 10
9individual/g; AMS is not less than 20U/g; Cellulose enzyme is not less than 10U/g; NSP complex enzyme is not less than 10U/g; Protease addition is not less than 100U/g; Be that 20%-50% adds fish molten slurry according to the mass ratio accounting for described basal diet gross mass;
(2) add water adjustment material water quality than being 1:0.7-1:1, control temperature is 30 DEG C of-50 DEG C of sealing and fermenting;
(3) fermented and adopted low temperature 60 DEG C oven dry.
2. the fermentation process adding the molten slurry of fish in corn-soybean meal diet according to claim 1, is characterized in that: described bacillus amyloliquefaciens HFJ-7 is bacillus amyloliquefaciens
bacillus amyloliquefaciens, its deposit number is CGMCC No. 10011.
3. the fermentation process adding the molten slurry of fish in corn-soybean meal diet according to claim 1, is characterized in that: the preparation process of the bacillus amyloliquefaciens HFJ-7 bacterium powder containing described bacillus amyloliquefaciens HFJ-7 is as follows:
(1) be inoculated into by bacillus amyloliquefaciens HFJ-7 and be equipped with in the sterilizing seeding tank of seed culture medium, cultivate 10h for 37 DEG C, form seed culture fluid, described seed culture medium component comprises: molasses, 24.5 ~ 25.0 g/L; Peptone, 13.5 ~ 13.7 g/L; Potassium dihydrogen phosphate, 1.7 ~ 1.8 g/L; Natrium citricum, 1.5 ~ 1.6 g/L; Ammonium nitrate, 1.5 ~ 1.6 g/L; Magnesium sulfate, 0.5 ~ 0.52 g/L; Regulate initial pH to 7.2;
(2) described seed culture fluid is forwarded to the production tank that fermentation medium is housed and carries out fermented and cultured, cultivate 24h, when spore rate is more than 95%, stop tank for 37 DEG C, obtain zymotic fluid, described fermentation medium composition comprises: 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; Dipotassium hydrogen phosphate, 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; Defoamer, 1.3 ~ 1.4 g/L; Regulate initial pH to 7.4, the bacterium amount of described bacillus amyloliquefaciens HFJ-7 zymotic fluid reaches 450 × 10
8cFU/g ~ 500 × 10
8cFU/g;
(3) carry out spraying dry after described zymotic fluid being added carrier, obtain bacteria containing amount 2000 × 10
8~ 2200 × 10
8the bacillus amyloliquefaciens HFJ-7 bacterium powder of CFU/g, described carrier is precipitated calcium carbonate, adds the mass ratio that the amount of carrier accounts for zymotic fluid to be: 10%-15%.
4. the biofermentation method adding the molten slurry of fish in corn-soybean meal diet according to claim 1, is characterized in that: in described corn-soybean meal basal diet, the mass ratio of corn and dregs of beans is 4:1-3:2.
5. the biofermentation method adding the molten slurry of fish in corn-soybean meal diet according to claim 1, is characterized in that: the protease added in described step (1) is by enzyme addition alkali protease: neutral proteinase: the mixture of three kinds of components of acid protease=2:5:3.
6. the biofermentation method adding the molten slurry of fish in corn-soybean meal diet according to claim 1, is characterized in that: sealing and fermenting 48h in described step (2), aerobic fermentation 12h in sealing and fermenting, anaerobic fermentation 36h.
7. the biofermentation method adding the molten slurry of fish in corn-soybean meal diet according to claim 3, is characterized in that: in described step (3), spray-dired EAT is 145 DEG C-150 DEG C, and leaving air temp is 65 DEG C-70 DEG C.
8. the fermentate that the fermentation process described in any one of claim 1-7 is obtained.
9. fermentate according to claim 8, is characterized in that: the pH3-5 of described fermentate, lactic acid content 10-20%, and little peptide content 20-30%, viable count is up to 9 × 10
10cfu/g ~ 3.8 × 10
12cfu/g.
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| CN104872379B (en) | 2018-07-24 |
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