CN115606681A - Broussonetia papyrifera leaf compound fermentation feed and preparation method thereof - Google Patents
Broussonetia papyrifera leaf compound fermentation feed and preparation method thereof Download PDFInfo
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- CN115606681A CN115606681A CN202211376315.7A CN202211376315A CN115606681A CN 115606681 A CN115606681 A CN 115606681A CN 202211376315 A CN202211376315 A CN 202211376315A CN 115606681 A CN115606681 A CN 115606681A
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- broussonetia papyrifera
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A23K—FODDER
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- A23K20/30—Oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
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Abstract
The invention discloses a paper mulberry leaf compound fermented feed. The invention discloses a preparation method of the broussonetia papyrifera leaf compound fermented feed, which comprises the following steps: (1) Crushing and sieving broussonetia papyrifera leaves, sterilizing by high-pressure steam, cooling to room temperature, adding water, uniformly stirring, inoculating a first microbial inoculum, fermenting for 10-20h in an anaerobic environment at 35-40 ℃, and discharging to obtain a primary fermentation material; (2) Sterilizing the primary fermented material with high pressure steam, cooling to room temperature, inoculating a second microbial inoculum, fermenting for 5-10h at 35-42 ℃ in an aerobic environment, and discharging to obtain a secondary fermented material; (3) Sterilizing the secondary fermentation material by high-pressure steam, cooling to room temperature, adding isobutyric acid, stirring uniformly, inoculating a third microbial inoculum, fermenting for 1-4h at 25-28 ℃ in an aerobic environment, discharging, adding corn, soybean meal, stone powder, vitamin mineral premix and corn oil, and mixing uniformly to obtain the broussonetia papyrifera leaf compound fermentation feed.
Description
Technical Field
The invention relates to the technical field of paper mulberry products, in particular to paper mulberry leaf compound fermentation feed and a preparation method thereof.
Background
Broussonetia papyrifera belongs to Broussonetia of Moraceae, is deciduous arbor, is mainly distributed in yellow river and southern region of China, fresh leaves of Broussonetia papyrifera are used for feeding pigs, fruits are used as medicines, and stems are commonly used for papermaking. Broussonetia papyrifera can benefit from its own high crude protein as an unconventional feed resource. The crude protein content in paper mulberry is slightly higher than that of alfalfa although not being as high as that of soybean meal, cottonseed meal and alfalfa in full-bloom stage.
At present, the broussonetia papyrifera is preliminarily applied to breeding of cattle, sheep, live pigs, poultry, rabbits and the like, and the urgent requirements of the breeding industry of China on green feed and protein feed sources are relieved to a certain extent. At present, the protein source of China still depends on import seriously, and meanwhile, in order to respond to the national comprehensive policy of 'banning resistance', the research of paper mulberry becomes one of the important subjects in the field of cultivation.
The broussonetia papyrifera has great development potential, but due to factors such as high content of self-fiber, the high-dosage addition can influence the digestion, absorption, growth and development of animals, and influences the enrichment of the animal self-flavor protein and the change of other biological functions. Furthermore, animals have different species and body types, and the addition mode of the broussonetia papyrifera is different, so that the feed additive is closely related to the difference of individual digestive systems of the animals and is also related to the tolerance of different animals to the feed of the broussonetia papyrifera.
Numerous studies have shown that poultry can utilize broussonetia papyrifera feed, but due to the structural limitation of poultry digestive tracts, the poultry has limited digestive capacity on the broussonetia papyrifera feed, so that the adding effect is not obvious. At present, the broussonetia papyrifera feed suitable for poultry is researched and established, and the healthy sustainable development of animal husbandry in China is further promoted.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a broussonetia papyrifera leaf compound fermented feed and a preparation method thereof.
A preparation method of a broussonetia papyrifera leaf composite fermented feed comprises the following steps:
(1) Crushing and sieving paper mulberry leaves, sterilizing by high-pressure steam, cooling to room temperature, adding water, stirring uniformly, inoculating a first microbial inoculum, fermenting for 10-20h in an anaerobic environment at 35-40 ℃, and discharging to obtain a primary fermentation material;
(2) Sterilizing the primary fermented material with high pressure steam, cooling to room temperature, inoculating a second microbial inoculum, fermenting for 5-10h at 35-42 ℃ in an aerobic environment, and discharging to obtain a secondary fermented material;
(3) Sterilizing the secondary fermentation material by high-pressure steam, cooling to room temperature, adding isobutyric acid, stirring uniformly, inoculating a third microbial inoculum, fermenting for 1-4h at 25-28 ℃ in an aerobic environment, discharging, adding corn, soybean meal, stone powder, vitamin mineral premix and corn oil, and mixing uniformly to obtain the broussonetia papyrifera leaf compound fermentation feed.
Preferably, in the step (1), the first microbial agent is a bacillus subtilis microbial agent, and the concentration of viable bacteria is more than or equal to 3.0 multiplied by 10 8 cfu/mL。
Preferably, the bacillus subtilis preparation comprises: the N-8 strain and the J-4 strain.
Preferably, in the step (2), the mass ratio of the broussonetia papyrifera leaves to the first microbial inoculum is 10-20:1-2.
Preferably, in step (2), the second microbial inoculum comprises geotrichum candidum, lactic acid bacteria and aspergillus niger, and the concentration of total viable bacteria is more than or equal to 1.0 multiplied by 10 8 cfu/mL。
Preferably, in the step (2), the mass ratio of the primary fermentation material to the second microbial inoculum is 15-50:0.1-1.
Preferably, in the step (3), the third microbial inoculum is a Eurotium cristatum microbial inoculum, and the concentration of viable bacteria is more than or equal to 1.0 multiplied by 10 8 cfu/mL。
Preferably, in the step (3), the mass ratio of the secondary fermentation material, the isobutyric acid and the third microbial inoculum is 20-30:1-2:0.1-0.5.
Preferably, in the step (3), the mass ratio of the secondary fermentation material, the corn, the soybean meal, the stone powder, the vitamin and mineral premix and the corn oil is 20-30:60-65:22-26:6-10:4-6:0.3-0.7.
A broussonetia papyrifera leaf compound fermented feed is prepared by adopting the preparation method of the broussonetia papyrifera leaf compound fermented feed.
The broussonetia papyrifera leaf compound fermented feed is applied to feeding of laying hens.
The technical effects of the invention are as follows:
in the step (1), through the biotransformation function of the first microbial inoculum, the secreted protease can effectively degrade and convert crude protein in the broussonetia papyrifera leaves into small molecular peptides and free amino acids; in the step (2), the influence of the first microbial inoculum on the subsequent fermentation process can be effectively eliminated through high-temperature inactivation, experiments show that lysine, histidine, phenylalanine, leucine, alanine, glycine and proline in the microbial inoculum are obviously increased in different degrees, if the first microbial inoculum continuously participates in the second-stage fermentation, the ratio of arginine/lysine in the system is increased from 1.1-1.5 to 3.01-3.25 due to protease generated by the first microbial inoculum, and the ideal ratio of arginine/lysine cannot be achieved, so that the production performance of poultry is further caused.
In the step (2), geotrichum candidum, lactic acid bacteria and aspergillus niger are compounded, so that the geotrichum candidum can quickly grow on a primary fermentation material, a large amount of protein, fat, vitamins, nucleic acid and the like are produced, the three strains are synergistic under the same fermentation condition, the strains are complementary, and secondary full fermentation of the broussonetia papyrifera leaves is effectively completed.
The eurotium cristatum preparation is used as a strain in the Fuzhuan tea flowering process, related reports are not seen when the eurotium cristatum preparation is used for feed fermentation at present, and as the special intestinal tract of laying hens is short in digestive tract and short in chyme retention time, isobutyric acid is added and is matched with a third microbial inoculum for fermentation in the three-time fermentation process, the product can effectively enhance the digestion and absorption capacity of the intestinal tract of the laying hens on nutrient substances, the intestinal mucosa immune function can be improved, the gastrointestinal tract function of the laying hens can be effectively improved, and the palatability and the storage resistance of the feed can be effectively improved.
The applicant finds that the specific gravity of the eggshell of the laying hen is large due to the addition of the unfermented broussonetia papyrifera leaves in the daily ration, and the adoption of the broussonetia papyrifera leaf composite fermented feed not only increases the weight of the eggshell, but also effectively deepens the color of the yolk. Meanwhile, the three-stage fermentation of the method has irreplaceability, scientific research personnel try to change the sequence of the step-by-step fermentation, or two strains are used for fermentation together, and the final fermentation depth is greatly influenced. By adopting the fermentation process, the final fermented feed can promote the growth of the laying hens, and particularly has obvious promotion effect on increasing the growth and production performance of the laying hens in the egg producing period.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples.
The vitamin mineral premix used comprises: calcium hydrogen phosphate, sodium chloride, stone powder, feed grade lysine, feed grade methionine, and feed grade vitamins (A, D) 3 、E、K 3 、B 1 、B 2 、B 6 、B 12 Biotin, folic acid, nicotinic acid, pantothenic acid, and choline chloride), feed-grade trace elements, phytase; VA:120000-200000IU/kg, VD 3 ：30000-70000IU/kg，VE：≥320mg/kg，K 3 ：≥20mg/kg，VB 2 : not less than 90mg/kg, niacin: 420mg/kg or more, fe:600-4400mg/kg, cu:80-550mg/kg, zn:900-2400mg/kg, mn:1000-3000mg/kg.
Example 1
A preparation method of a broussonetia papyrifera leaf compound fermentation feed comprises the following steps:
(1) Pulverizing 10kg folium Broussonetiae, sieving with 18 mesh sieve, sterilizing at 105 deg.C under high pressure steam for 1min, cooling to room temperature, adding 5kg water, stirring, inoculating 1kg viable bacteria with concentration of 3.0 × 10 8 cfu/mL of Bacillus subtilisFermenting the bacillus agent for 10h in an anaerobic environment at the temperature of 35 ℃, and discharging to obtain a primary fermentation material.
The bacillus subtilis microbial inoculum comprises an N-8 strain and a J-4 strain, wherein the viable bacteria concentration ratio of the two strains is 3:2.
(2) Sterilizing 15kg of primary fermented material at 110 deg.C for 10s with high pressure steam, cooling to room temperature, inoculating 0.1kg of viable bacteria with concentration not less than 1.0 × 10 8 And (3) fermenting the cfu/mL second microbial inoculum for 5h in an aerobic environment at the temperature of 35 ℃, and discharging to obtain a secondary fermentation material.
The second microbial inoculum is prepared from geotrichum candidum, lactic acid bacteria and aspergillus niger according to the viable bacteria concentration ratio of 1:3: 1.
(3) Sterilizing 20kg of secondary fermentation material with high pressure steam at 110 deg.C for 5s, cooling to room temperature, adding 1kg of isobutyric acid, stirring, inoculating 0.1kg of viable bacteria with concentration not less than 1.0 × 10 8 cfu/mL of the Eurotium cristatum is fermented for 1h in an aerobic environment at the temperature of 25 ℃, and then the mixture is discharged, and 60kg of corn, 22kg of bean pulp, 6kg of stone powder, 4kg of vitamin mineral premix and 0.3kg of corn oil are added and mixed uniformly to obtain the paper mulberry leaf compound fermented feed.
Example 2
A preparation method of a broussonetia papyrifera leaf composite fermented feed comprises the following steps:
(1) Pulverizing 20kg folium Broussonetiae, sieving with 18 mesh sieve, sterilizing at 115 deg.C under high pressure steam for 3min, cooling to room temperature, adding 30kg water, stirring, inoculating 2kg viable bacteria with concentration of 3.0 × 10 8 cfu/mL of a bacillus subtilis microbial inoculum, fermenting for 20h in an anaerobic environment at the temperature of 40 ℃, and discharging to obtain a primary fermentation material.
The bacillus subtilis microbial inoculum comprises an N-8 strain and a J-4 strain, wherein the viable bacteria concentration ratio of the two strains is 4:3.
(2) Sterilizing 50kg of primary fermented material at 120 deg.C with high pressure steam for 30s, cooling to room temperature, inoculating 1kg of viable bacteria with concentration of 1.0 × 10 or more 8 cfu/mL of second microbial inoculum is fermented for 10 hours in an aerobic environment at the temperature of 42 ℃, and the material is discharged to obtain a secondary fermentation material.
The second microbial inoculum is prepared from geotrichum candidum, lactic acid bacteria and aspergillus niger according to the viable bacteria concentration ratio of 2:5:3, and (3).
(3) Sterilizing 30kg of secondary fermentation material at 120 deg.C with high pressure steam for 30s, cooling to room temperature, adding 2kg of isobutyric acid, stirring, inoculating 0.5kg of viable bacteria with concentration not less than 1.0 × 10 8 cfu/mL of the Eurotium cristatum is fermented for 4 hours at 28 ℃ in an aerobic environment, and then discharged, 65kg of corn, 26kg of soybean meal, 10kg of stone powder, 6kg of vitamin mineral premix and 0.7kg of corn oil are added and mixed uniformly to obtain the broussonetia papyrifera leaf compound fermented feed.
Example 3
A preparation method of a broussonetia papyrifera leaf compound fermentation feed comprises the following steps:
(1) Pulverizing 13kg folium Broussonetiae, sieving with 18 mesh sieve, sterilizing with high pressure steam at 112 deg.C for 1.5min, cooling to room temperature, adding 20kg water, stirring, inoculating 1.3kg viable bacteria with concentration of 3.0 × 10 or more 8 cfu/mL of a bacillus subtilis microbial inoculum is fermented for 13h at 38 ℃ in an anaerobic environment, and the material is discharged to obtain a primary fermentation material.
The bacillus subtilis microbial inoculum comprises an N-8 strain and a J-4 strain, wherein the viable bacteria concentration ratio of the N-8 strain to the J-4 strain is 3.7:2.2.
(2) Sterilizing 40kg of primary fermented material with high pressure steam at 112 deg.C for 25s, cooling to room temperature, inoculating 0.4kg of viable bacteria with concentration not less than 1.0 × 10 8 And (3) fermenting the cfu/mL second microbial inoculum for 6h in an aerobic environment at the temperature of 40 ℃, and discharging to obtain a secondary fermentation material.
The second microbial inoculum is prepared from geotrichum candidum, lactic acid bacteria and aspergillus niger according to the viable bacteria concentration ratio of 1.7:3.5: 2.5.
(3) Sterilizing 22kg of secondary fermentation material at 117 deg.C with high pressure steam for 15s, cooling to room temperature, adding 1.7kg of isobutyric acid, stirring, inoculating 0.2kg of viable bacteria with concentration not less than 1.0 × 10 8 cfu/mL of the Eurotium cristatum is fermented for 2h in an aerobic environment at the temperature of 27 ℃, discharged, and added with 64kg of corn, 24kg of soybean meal, 9kg of stone powder, 4.5kg of vitamin mineral premix and 0.6kg of corn oil to be uniformly mixed to obtain the broussonetia papyrifera leaf compound fermented feed.
Example 4
A preparation method of a broussonetia papyrifera leaf compound fermentation feed comprises the following steps:
(1) Pulverizing 1kg folium Broussonetiae, and sieving with 18 mesh sieveSieving, sterilizing with high pressure steam at 109 deg.C for 2min, cooling to room temperature, adding 18kg water, stirring, inoculating 1.5kg viable bacteria with concentration of 3.0 × 10 or more 8 cfu/mL of a bacillus subtilis microbial inoculum, fermenting for 15h in an anaerobic environment at the temperature of 38 ℃, and discharging to obtain a primary fermentation material.
The bacillus subtilis microbial inoculum comprises an N-8 strain and a J-4 strain, wherein the viable bacteria concentration ratio of the two strains is 3.5:2.6.
(2) Sterilizing 35kg of primary fermented material at 116 deg.C with high pressure steam for 20s, cooling to room temperature, inoculating 0.6kg of viable bacteria with concentration not less than 1.0 × 10 8 And (3) fermenting the cfu/mL second microbial inoculum for 7h in an aerobic environment at the temperature of 38 ℃, and discharging to obtain a secondary fermentation material.
The second microbial inoculum is prepared from geotrichum candidum, lactic acid bacteria and aspergillus niger according to the viable bacteria concentration ratio of 1.6:38: 1.8.
(3) Sterilizing 26kg of secondary fermentation material at 115 deg.C with high pressure steam for 18s, cooling to room temperature, adding 1.6kg of isobutyric acid, stirring, inoculating 0.3kg of viable bacteria with concentration not less than 1.0 × 10 8 cfu/mL of the Eurotium cristatum is fermented for 2h at 26 ℃ in an aerobic environment, and 63kg of corn, 24kg of soybean meal, 8kg of stone powder, 4.8kg of vitamin mineral premix and 0.5kg of corn oil are added and mixed uniformly to obtain the broussonetia papyrifera leaf compound fermented feed.
Example 5
A preparation method of a broussonetia papyrifera leaf compound fermentation feed comprises the following steps:
(1) Pulverizing folium Broussonetiae 15.5kg, sieving with 18 mesh sieve, sterilizing with high pressure steam at 110 deg.C for 2min, cooling to room temperature, adding 18kg water, stirring, inoculating 1.5kg viable bacteria with concentration of 3.0 × 10 or more 8 cfu/mL of a bacillus subtilis microbial inoculum, fermenting for 15h in an anaerobic environment at the temperature of 37 ℃, and discharging to obtain a primary fermentation material.
The bacillus subtilis microbial inoculum comprises an N-8 strain and a J-4 strain, wherein the viable bacteria concentration ratio of the two strains is 3.5:2.5.
(2) Sterilizing 33kg of primary fermented material at 115 deg.C with high pressure steam for 20s, cooling to room temperature, inoculating 0.6kg of viable bacteria with concentration of 1.0 × 10 or more 8 cfu/mL of second microbial inoculum is fermented for 7 hours in an aerobic environment at the temperature of 39 ℃,discharging to obtain the secondary fermentation material.
The second microbial inoculum is prepared from geotrichum candidum, lactic acid bacteria and aspergillus niger according to the viable bacteria concentration ratio of 1.5:4: 2.
(3) Sterilizing 25kg of secondary fermentation material at 115 deg.C under high pressure steam for 18s, cooling to room temperature, adding 1.5kg of isobutyric acid, stirring, inoculating 0.3kg of viable bacteria with concentration not less than 1.0 × 10 8 cfu/mL of the Eurotium cristatum, fermenting for 2.5h at 26.5 ℃ in an aerobic environment, discharging, adding 62.3kg of corn, 24.3kg of soybean meal, 7.97kg of stone powder, 4.93kg of vitamin mineral premix and 0.5kg of corn oil, and uniformly mixing to obtain the broussonetia papyrifera leaf compound fermented feed.
The nutrition level of the obtained broussonetia papyrifera leaf compound fermentation feed is tested and calculated, and the method specifically comprises the following steps: the metabolizable energy is 13.57MJ/kg, the crude protein content is 20.03 percent, the lysine content is 1.46 percent, the methionine content is 0.75 percent, the calcium element content is 2.56 percent, and the total phosphorus content is 0.41 percent.
Comparative example 1
A preparation method of a broussonetia papyrifera leaf composite fermented feed comprises the following steps:
(1) Pulverizing folium Broussonetiae 15.5kg, sieving with 18 mesh sieve, sterilizing with high pressure steam at 110 deg.C for 2min, cooling to room temperature, adding 18kg water, stirring, inoculating 1.5kg viable bacteria with concentration of 3.0 × 10 8 cfu/mL of a bacillus subtilis microbial inoculum, fermenting for 15h in an anaerobic environment at the temperature of 37 ℃, and discharging to obtain a primary fermentation material.
The bacillus subtilis microbial inoculum comprises an N-8 strain and a J-4 strain, wherein the concentration ratio of viable bacteria of the N-8 strain to the J-4 strain is 3.5:2.5.
(2) Sterilizing 33kg of primary fermented material at 115 deg.C with high pressure steam for 20s, cooling to room temperature, inoculating 0.6kg of viable bacteria with concentration of 1.0 × 10 or more 8 And (3) fermenting the cfu/mL second microbial inoculum for 7h in an aerobic environment at the temperature of 39 ℃, and discharging to obtain a secondary fermentation material.
The second microbial inoculum is prepared from geotrichum candidum, lactic acid bacteria and aspergillus niger according to the viable bacteria concentration ratio of 1.5:4: 2.
(3) Sterilizing 25kg of secondary fermentation material at 115 deg.C under high pressure steam for 18s, cooling to room temperature, adding 62.3kg of corn, 24.3kg of soybean meal, 7.97kg of stone powder, 4.93kg of vitamin mineral premix and 0.5kg of corn oil, and mixing to obtain the final product.
Comparative example 2
A preparation method of a broussonetia papyrifera leaf compound fermentation feed comprises the following steps:
(1) Pulverizing folium Broussonetiae 15.5kg, sieving with 18 mesh sieve, sterilizing with high pressure steam at 110 deg.C for 2min, cooling to room temperature, adding 18kg water, stirring, inoculating 1.5kg viable bacteria with concentration of 3.0 × 10 8 cfu/mL of the bacillus subtilis microbial inoculum is fermented for 15 hours at 37 ℃ in an anaerobic environment, and the material is discharged to obtain a primary fermentation material.
The bacillus subtilis microbial inoculum comprises an N-8 strain and a J-4 strain, wherein the viable bacteria concentration ratio of the two strains is 3.5:2.5.
(2) Sterilizing 33kg of primary fermentation material at 115 deg.C with high pressure steam for 20s, cooling to room temperature, adding 1.5kg of isobutyric acid, stirring, inoculating 0.6kg of viable bacteria with concentration not less than 1.0 × 10 8 cfu/mL second microbial inoculum and 0.3kg viable bacteria concentration more than or equal to 1.0 multiplied by 10 8 cfu/mL of the Eurotium cristatum, fermenting for 9.5h at 26.5 ℃ in an aerobic environment, discharging, adding 62.3kg of corn, 24.3kg of soybean meal, 7.97kg of stone powder, 4.93kg of vitamin mineral premix and 0.5kg of corn oil, and uniformly mixing to obtain the broussonetia papyrifera leaf compound fermented feed.
The second microbial inoculum is prepared from geotrichum candidum, lactic acid bacteria and aspergillus niger according to the viable bacteria concentration ratio of 1.5:4: 2.
Comparative example 3
A preparation method of a broussonetia papyrifera leaf composite fermented feed comprises the following steps:
(1) Pulverizing folium Broussonetiae 15.5kg, sieving with 18 mesh sieve, sterilizing with high pressure steam at 110 deg.C for 2min, cooling to room temperature, adding 18kg water, stirring, inoculating 1.5kg viable bacteria with concentration of 3.0 × 10 or more 8 cfu/mL of a bacillus subtilis microbial inoculum, fermenting for 15h in an anaerobic environment at the temperature of 37 ℃, and discharging to obtain a primary fermentation material.
The bacillus subtilis microbial inoculum comprises an N-8 strain and a J-4 strain, wherein the viable bacteria concentration ratio of the two strains is 3.5:2.5.
(2) Sterilizing 33kg of the primary fermented material with high pressure steam at 115 deg.CCooling to room temperature for 20s, inoculating 0.6kg viable bacteria with concentration not less than 1.0 × 10 8 cfu/mL of second microbial inoculum is fermented for 7 hours in an aerobic environment at the temperature of 39 ℃, and the secondary fermentation material is obtained after discharging.
The second microbial inoculum is prepared from geotrichum candidum, lactic acid bacteria and aspergillus niger according to the viable bacteria concentration ratio of 1.5:4: 2.
(3) Sterilizing 25kg of secondary fermentation material at 115 deg.C under high pressure steam for 18s, cooling to room temperature, inoculating 0.3kg of viable bacteria with concentration not less than 1.0 × 10 8 cfu/mL of the Eurotium cristatum, fermenting for 2.5h at 26.5 ℃ in an aerobic environment, discharging, adding 62.3kg of corn, 24.3kg of soybean meal, 7.97kg of stone powder, 4.93kg of vitamin mineral premix and 0.5kg of corn oil, and uniformly mixing to obtain the broussonetia papyrifera leaf compound fermented feed.
The fermented feed obtained in example 5 and comparative examples 1-3 was used for the group feeding experiment of laying hens as follows: the experiment is carried out in a certain large-scale chicken farm with Anhui fattener, the experiment selects the Hailan brown laying hens in the healthy laying period of 23 weeks, the temperature of the henhouse in the experiment period is 28 +/-2 ℃, the relative humidity is 55-62%, the experiment is divided into 4 groups, each group is 7, and each group is 12 chickens; pre-run period 7d, positive run period 54d. Observing the health condition of the laying hens every day, and timely making test records, wherein the test data are processed by EXCEL2007, the test results are represented by the average value +/-standard error, and P <0.05 is used as a judgment standard with obvious difference.
1. Measurement of production index
During the test period, the total egg production, the total egg weight, the feed intake and the qualified egg number are repeatedly recorded every day, and the average daily feed intake, the average egg weight, the egg production rate and the qualified egg rate of each group are counted by taking the group as a unit, wherein: average egg weight = total egg weight (g)/egg production number; laying rate = number of eggs/number of chickens × 100%; the qualified egg rate = the number of qualified eggs/the total number of eggs × 100%.
The test results are shown in table 1 below:
TABLE 1 layer performance
As can be seen from Table 1, compared with comparative examples 1-3, the average daily food intake and the average egg weight of the invention are both significantly increased (P is less than 0.05); in particular, example 5 significantly increased the average egg weight (P < 0.05) compared to comparative example 1. The final fermented feed can promote the growth of the laying hens, and particularly has obvious effect of promoting the growth and the production performance of the laying hens in an egg producing period.
2. Egg quality determination
Collecting egg samples one day before the test, randomly selecting 4 eggs with perfect appearance and approximate weight every time repeating, and weighing the weight of each egg, the length and the short diameter of each egg, the eggshell strength, the egg white height and the eggshell thickness (average value of blunt end, tip end and middle part);
the yolk color is scored by using a poultry yolk colorimetric fan (Luchuan creature), the yolk and the egg white are separated, the weight of the yolk, the weight of the egg white and the weight of the eggshell are respectively weighed, and the specific gravity of the yolk, the egg white and the eggshell is respectively calculated. The test results are shown in table 2 below:
TABLE 2 egg quality
As can be seen from Table 2, the present invention significantly darkens the color of the yolk (P < 0.05) compared to the comparative examples 1-3; obviously reduces the specific gravity and thickness of the eggshell (P < 0.05).
3. Intestinal index of laying hen
Alpha-amylase: measured by a starch-iodine colorimetric method, purchased from Nanjing to build a Biotech Co., ltd., a product number C016-1-1; trypsin: measured by an ultraviolet colorimetric method, purchased from Nanjing to build a Biotech Co., ltd, with the product number of A080-2-2; lipase: measured by a colorimetric method, purchased from Nanjing to build a Biotech Co., ltd, a product number A054-1-1; total protein: measured by Coomassie brilliant blue method, purchased from Nanjing to build Biotech Co., ltd, article number A045-2-2; chicken secreted immunoglobulin: the kit is used for determination by an ELISA detection kit, and is purchased from Shanghai enzyme-linked biotechnology, inc., with the commodity number ml002778.
The test results are shown in table 3 below:
TABLE 3 intestinal tract index of laying hens
As can be seen from Table 3, at week 8 of the experiment, the activity of the hen jejunum tissue lipase of the present invention was significantly higher than that of the comparative example (P < 0.01), while the activity of the hen jejunum tissue amylase was significantly higher than that of the comparative example (P < 0.01) compared to that of comparative example 1.
4. Egg chicken jejunum villus form
Performing histomorphology observation on the laying hen intestinal tract sample through an optical microscope; the villus height and crypt depth of each group of sample slices were measured, 3 replicates per slice were measured, and the villus height/crypt depth was calculated.
Wherein the height of the villus is the length from the top of the villus to the entrance of the recess, and the depth of the recess is the length from the base of the villus to the submucosa. The test results are shown in table 4 below:
TABLE 4 intestinal tract morphology of layer chicken
As can be seen from Table 4, at week 8 of the experiment, the ratio of the length of jejunal villus to the depth of crypt of the laying hens of the invention was significantly higher than that of the comparative example (P < 0.05). The invention can effectively enhance the digestive absorption capacity of the intestinal tract of the laying hens to nutrient substances, improve the intestinal mucosa immunity function and effectively improve the gastrointestinal tract function of the laying hens.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered as the technical solutions and the inventive concepts of the present invention within the technical scope of the present invention.
Claims (10)
1. A preparation method of a broussonetia papyrifera leaf compound fermentation feed is characterized by comprising the following steps:
(1) Crushing and sieving broussonetia papyrifera leaves, sterilizing by high-pressure steam, cooling to room temperature, adding water, uniformly stirring, inoculating a first microbial inoculum, fermenting for 10-20h in an anaerobic environment at 35-40 ℃, and discharging to obtain a primary fermentation material;
(2) Sterilizing the primary fermented material with high pressure steam, cooling to room temperature, inoculating second microbial inoculum, fermenting at 35-42 deg.C under aerobic environment for 5-10h, and discharging to obtain secondary fermented material;
(3) Sterilizing the secondary fermentation material by high-pressure steam, cooling to room temperature, adding isobutyric acid, stirring uniformly, inoculating a third microbial inoculum, fermenting for 1-4h at 25-28 ℃ in an aerobic environment, discharging, adding corn, soybean meal, stone powder, vitamin mineral premix and corn oil, and mixing uniformly to obtain the broussonetia papyrifera leaf compound fermentation feed.
2. The preparation method of the broussonetia papyrifera leaf compound fermented feed according to claim 1, wherein in the step (1), the first microbial agent is a bacillus subtilis microbial agent, and the concentration of viable bacteria is more than or equal to 3.0 x 10 8 cfu/mL。
3. The preparation method of the broussonetia papyrifera leaf compound fermented feed as claimed in claim 2, wherein the bacillus subtilis microbial inoculum comprises: the N-8 strain and the J-4 strain.
4. The preparation method of the broussonetia papyrifera leaf compound fermented feed according to claim 2, wherein in the step (2), the mass ratio of the broussonetia papyrifera leaves to the first microbial inoculum is 10-20:1-2.
5. The preparation method of the broussonetia papyrifera leaf composite fermented feed according to claim 1, wherein in the step (2), the second microbial agent comprises geotrichum candidum, lactic acid bacteria and aspergillus niger, and the concentration of total viable bacteria is more than or equal to 1.0 x 10 8 cfu/mL。
6. The preparation method of the broussonetia papyrifera leaf compound fermented feed according to claim 5, wherein in the step (2), the mass ratio of the primary fermented material to the second microbial inoculum is 15-50:0.1-1.
7. The preparation method of the broussonetia papyrifera leaf compound fermented feed according to claim 1, wherein in the step (3), the third microbial agent is a eurotium cristatum microbial agent, and the concentration of viable bacteria is more than or equal to 1.0 x 10 8 cfu/mL。
8. The preparation method of the broussonetia papyrifera leaf compound fermented feed according to claim 7, wherein in the step (3), the mass ratio of the secondary fermented material to the isobutyric acid to the third microbial inoculum to the corn to the soybean meal to the stone powder to the vitamin and mineral premix to the corn oil is 20-30:1-2:0.1-0.5:60-65:22-26:6-10:4-6:0.3-0.7.
9. A broussonetia papyrifera leaf composite fermented feed is characterized by being prepared by the preparation method of the broussonetia papyrifera leaf composite fermented feed according to any one of claims 1 to 8.
10. The use of the broussonetia papyrifera leaf compound fermented feed as claimed in claim 9 in feeding laying hens.
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