CN114586890A - Method for preparing composite flora feed by utilizing juncao - Google Patents
Method for preparing composite flora feed by utilizing juncao Download PDFInfo
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Classifications
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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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
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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
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/121—Brevis
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- 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
- Y02P60/87—Re-use of by-products of food processing for fodder production
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Abstract
The invention discloses a method for preparing a compound flora feed by utilizing Juncao. The invention takes mushroom grass as a main material, corn flour and wheat bran as auxiliary materials, and adds white rot fungus enzyme liquid to degrade cellulose, hemicellulose and lignin, and simultaneously adds a composite microbial inoculum to carry out mixed fermentation, thereby obtaining the composite flora feed. The invention greatly shortens the time of grass silage fermentation, reduces the cellulose and lignin content which is difficult to digest and absorb by animals, and improves the nutritive value and palatability of grass feed; improving feed function, enhancing animal immunity and oxidation resistance; the health cultivation is beneficial to ensuring the food safety; the biological fermentation feed replaces part of protein feed, reduces the feed grain, replaces the grain with grass, expands the livestock industry and improves the economic benefit.
Description
Technical Field
The invention belongs to the technical field of feed processing, and particularly relates to a method for preparing a compound flora feed by utilizing Juncao.
Background
With the rapid development of animal husbandry, the shortage of feed resources is increasingly obvious, the protein feed raw materials such as corn, bean pulp, fish meal and the like are in shortage and have higher price, and the cheap miscellaneous meal cannot be fully utilized by animals and is greatly discarded, so that the resource waste is caused; secondly, part of nutrients which cannot be absorbed by organisms are discharged out of the body through the excrement, the treatment of the livestock excrement is improper, and the pollution to soil and atmosphere is caused; the abuse of antibiotics also enhances the drug resistance of pathogenic bacteria in livestock and poultry bodies, and the antibiotic residue of livestock products directly influences the food safety of human beings, thereby seriously influencing the health of the human beings. Healthy breeding, food shortage, environmental pollution, improvement of the flavor of animal products and the like are common problems in the livestock feed industry.
The biological fermentation feed is a new industry and provides a technical solution for transformation and upgrading of animal husbandry development. The Juncao is a perennial herbaceous plant of the Gramineae family, and mainly comprises plants of more than 20 genera, 46 species in total, such as the Gramineae family and the Libaike family. The species with wider planting area mainly comprise Jujun grass, oasis No. 1, oasis No. 2, oasis No. 3, oasis No. 5 and the like. The grass has thick and large leaves, soft and succulent leaves, is rich in nutrients such as saccharides, lignin, crude protein and crude fiber, has the advantages of good palatability, easy digestion and the like, and is a good choice for high-quality green feeds for poultry farming and aquaculture. The growth rate of the Juncao is fast, the yield is high, the stress resistance is strong, the high temperature resistance, drought resistance, salt resistance, cold resistance and the like, the application range of the Juncao is expanded, and the vacancy of low storage of the feed in winter in the north can be made up.
White rot fungus enzyme and mixed flora fermentation fungus grass are introduced to prepare the biological fermentation feed, so that the fermentation time of the fungus grass feed can be shortened, and the nutritive value and the palatability of the fungus grass feed are improved; improving intestinal flora of animals, improving immunity and oxidation resistance, and promoting nutrient absorption; healthy cultivation, food safety guarantee, development of livestock industry and improvement of economic benefits.
Currently, some researches on grass feed are carried out, but the researches are mostly limited to grass silage, so that the feed intake and the economic benefit of the feed are improved to a certain degree. However, the research on microbial fermentation of the grass feed is not many, and a microbial inoculum and a production process for biological fermentation of the grass feed with pertinence are not clear.
Disclosure of Invention
The invention aims to provide a method for preparing a compound flora feed by utilizing Juncao.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for preparing compound flora feed by using Juncao comprises taking Juncao as main material, taking corn flour and wheat bran as auxiliary materials, adding white-rot fungi crude enzyme solution to degrade cellulose, hemicellulose and lignin, and adding special compound flora for Juncao fermentation to perform mixed fermentation to obtain Juncao biological fermentation feed. The feed obtained by the method has obvious sour and wine flavors, soft and fluffy texture, golden color and rich nutrition; the production process is simple, short in time and low in cost, and is favorable for realizing industrial large-scale production of the grass-cultivated fermented feed.
The method for preparing the compound flora feed by using the juncao concretely comprises the following steps:
(1) harvesting fresh grass in the growth period of 2 months;
(2) respectively drying the harvested fresh Jujun grass, wheat bran and corn flour at 50 ℃ for 12 h, and crushing into 20-mesh fine powder for later use;
(3) uniformly mixing 80 parts of megatherium powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material;
(4) adding 0.05% (w/w) of white rot fungus crude enzyme liquid and 1.5-4.5% (v/w) of a grass-fungus fermentation composite microbial inoculum into a fermentation raw material, adding water to control the water content of the final fermentation raw material to be kept at 50-70%, removing air, compacting, sealing, and carrying out anaerobic fermentation at 25-30 ℃ in a dark place for 9 days to obtain the composite flora feed.
The growth period of the fresh grass is 2 months, at the time, the grass is about 2.5m high, the nutrient content is high, the content of crude protein is high, dry matter, crude fat and crude fiber reach high levels, and the digestion of animals is facilitated. The growth cycle is too short, and the dry matter and crude fiber content of the fungus grass are low; the growth cycle is too long, and the crude protein content of the juncao is low.
The preparation method of the white-rot fungi crude enzyme solution comprises the following steps: inoculating single colony of white rot fungi into a PDA solid culture medium, performing static culture at 28 ℃ for 5 days, taking 4 hypha blocks with the diameter of 5mm, transferring the hypha blocks into 200 mL of PDB liquid culture medium, performing culture at 28 ℃ and 160rpm/min for 5 days, forming hypha balls with the diameter of 1-3mm in the culture solution by utilizing the self-flocculation characteristic of the white rot fungi, and filtering the culture solution containing the hypha balls by using 4 layers of sterile gauze to obtain filtrate, namely the crude white rot fungi enzyme solution. Wherein, the addition amount of the white rot fungus enzyme production liquid is 0.5 g per kilogram of fermentation substrate.
The preparation method of the fungus grass fermentation composite bacteria comprises the following steps: uniformly mixing 90g of brown sugar and 1kg of sterile water, decocting, cooling to 30 ℃, adding 2.5g of lactobacillus plantarum strain, 2g of lactobacillus brevis strain, 1.2g of lactobacillus sakakii strain, 1g of lactobacillus paracasei strain, 1g of abnormal vekhem yeast strain, 0.8g of lactobacillus buchneri strain, 0.25g of bacillus subtilis strain and 0.25g of coprinus pratense strain, uniformly stirring, putting into a closed fermentation container, keeping the fermentation temperature at 30 ℃, and continuously fermenting for 1-5 days to obtain a fermented bacterial liquid; adding 2.64% (w/w) of zymocyte liquid into 20kg of fermentation substrate culture medium, adding water to control the water content of the final fermentation raw material to be kept at 60%, removing air, compacting and sealing, carrying out anaerobic fermentation at 28 ℃ in a dark place, taking 4 days as a fermentation period, and carrying out continuous passage for four times to obtain the composite bacterial agent for fermenting the bacteria and the grass.
The fermentation substrate culture medium is prepared by mixing 16kg of megaterium powder, 2kg of wheat bran powder and 2kg of corn powder.
The total viable bacteria concentration in the fermented bacterial liquid is 1 × 108cfu/g, wherein the number ratio of the lactobacillus plantarum, the lactobacillus brevis, the lactobacillus japonicas, the lactobacillus paracasei, the han-jiu yeast abnormal, the lactobacillus buchneri, the bacillus subtilis and the coprinus pratense is 43: 35: 14: 6: 5: 4: 1.
The total viable bacteria concentration in the composite bacterial agent for fermenting the grass is 2 multiplied by 109cfu/g, which mainly contains beneficial bacteria groups with the following relative abundance: pantoea genus (Pantoea51.57%), Lactobacillus (L.) genusLactobacillus21.76%), Pediococcus (Pediococcus0.01%), enterococcus (E), and B), (E), (Enterococcus0.02%), Derlichia zeylanica (R) ((R)Naganishia1.68%), Pichia pastoris (Meyerozyma，5.31%）。
The invention has the following remarkable advantages:
first, white rot fungi are the general term for filamentous fungi which are saprophytic on trees or wood and cause white rot of wood, and as the most effective and main lignin-degrading microorganisms, white rot fungi can not only completely degrade lignin into CO2And H2O, can also degrade hemicellulose and cellulose in a large amount and plays a key role in the carbon circulation of the nature; besides the capability of degrading complex organic matters, part of the white-rot fungi can also generate sporocarp which is rich in nutrition and even has medicinal and health-care values, so the white-rot fungi have extremely high application value in important fields closely related to human life, such as industry, agriculture and animal husbandry, food, medicine, environmental remediation and the like. The Jujun grass and oasis I contain a large amount of lignocellulose, and the invention uses white-rot fungi crude enzyme liquid which can be fermentedThe lignocellulose in the fermentation raw material is effectively degraded in the process.
The composite bacterial agent for fermenting the bacteria and the grass, which is adopted by the invention, contains various beneficial bacteria of lactobacillus plantarum, lactobacillus brevis, lactobacillus japonicas, lactobacillus paracasei, saccharomycetes, lactobacillus buchneri, bacillus subtilis and coprinus pusillii, and all the bacteria are fermented in a synergistic manner, so that macromolecular protein or polysaccharide in raw materials can be effectively degraded, the protein content is improved, the palatability is good, meanwhile, the storage time of the feed can be prolonged by the composite bacterial group for fermenting, the bacterial group can rapidly grow and reproduce and play the function of the bacterial group after entering the digestive intestinal tracts of animals, the composite bacterial agent has the function of resisting pathogenic microorganisms, and can produce antibacterial substances, inhibit the reproduction of harmful bacteria, promote the growth and development of the animals, maintain the balance of the intestinal flora of the animals, and enhance the immunity of the animals so as to prevent intestinal diseases.
The fermented feed of the fungi and grass has obvious sour and fragrant flavor and wine aroma, is soft and fluffy in texture, golden yellow in color, rich in nutrition, good in palatability, favorable for digestion and absorption, mellow in sour and sweet and unique in flavor; moreover, the process production of the grass-cultivated fermented feed is simple, the fermentation time is shortened, the cost is low, and the industrial large-scale production of the novel grass-cultivated fermented feed is favorably realized.
And the research on the utilization of the grass-fungus fermented feed at home and abroad is less, the research is mostly limited to grass-fungus silage, the fermentation time is long, and the grass-fungus fermented feed is not easy to store.
Detailed Description
In order to make the present invention more comprehensible, the technical solutions of the present invention are further described below with reference to specific embodiments, but the present invention is not limited thereto.
The white rot fungi, lactobacillus plantarum, lactobacillus brevis, lactobacillus jaborandi, lactobacillus paracasei, morehm vulgare, lactobacillus buchneri, bacillus subtilis and coprinus pusillii used in the following examples are all commercially available conventional existing strains or strains obtained by conventional screening, do not relate to the development of new strains, and only relate to the application of the strains. For example, the lactobacillus plantarum used in the following examples may specifically be a lactobacillus plantarum deposited in the china common microbiological collection center with the collection number of CGMCC NO: 1.12974; the lactobacillus brevis can be specifically a lactobacillus brevis strain with the preservation number of China general microbiological preservation management center of CGMCC NO: 1.2028; the American tiger lactobacillus can be specifically a strain with the preservation number of CGMCC NO: 1.2826; the Lactobacillus paracasei can be strain with commercial product number JCM1597 of Ningbo Ming boat Biotech Co., Ltd; the abnormal Weikehan yeast can be specifically the one with the preservation number of China general microbiological preservation management center of CGMCC NO: 2.1031; the lactobacillus buchneri can be specifically a lactobacillus buchneri strain with the preservation number of China general microbiological preservation management center of CGMCC NO: 1.15607; the bacillus subtilis can be specifically a bacillus subtilis with the preservation number of China general microbiological culture Collection center of CGMCC NO: 1.821 of a strain; specifically, the C.putida may be a strain having a commercial product number of Bio-103890, manufactured by Beijing Baiohbowei Biotechnology Co.
Example 1
Inoculating single colony of white rot fungi into a PDA (potato dextrose agar) solid culture medium, performing static culture at 28 ℃ for 5 days, taking 4 hypha blocks with the diameter of 5mm, transferring the hypha blocks into 200 mL of PDB liquid culture medium, performing culture at 28 ℃ and 160rpm/min for 5 days, and forming hypha balls with the diameter of 1-3mm in the culture solution by utilizing the self-flocculation characteristic of the white rot fungi to obtain a culture solution containing the hypha balls, namely a white rot fungi crude enzyme solution 1; filtering the white-rot fungi crude enzyme solution 1 with 4 layers of sterile gauze to obtain a filtrate, namely the white-rot fungi crude enzyme solution 2.
Example 2 preparation of a composite bacterial agent for fermentation of grass
The strain is as follows: lactobacillus plantarum, Lactobacillus brevis, Lactobacillus japonicas, Lactobacillus paracasei, Saccharomyces carlsbergensis, Lactobacillus buchneri, Bacillus subtilis, and Bacillus putrescentiae.
Fermentation substrate culture medium: 16kg of megaterium powder, 2kg of wheat bran powder and 2kg of corn powder. The preparation method of the pennisetum hydridum powder comprises the following steps: harvesting fresh Jujun grass with 2 months of growth period, drying the harvested fresh Jujun grass at 50 deg.C for 12 h, and pulverizing into 20 mesh fine powder for use.
Mixing brown sugar and sterile water according to a ratio of 90 g: 1kg of the raw materials are uniformly mixed, boiled and cooled to 30 ℃, then 2.5g of lactobacillus plantarum strain, 2g of lactobacillus brevis strain, 1.2g of lactobacillus japonicas strain, 1g of lactobacillus paracitrate strain, 1g of abnormal yeast Weikeham, 0.8g of lactobacillus buchneri strain, 0.25g of bacillus subtilis strain and 0.25g of coprinus pratense strain are added into the mixture, the mixture is uniformly stirred and then is put into a closed fermentation container, the fermentation temperature is kept at 30 ℃, the mixture is continuously fermented for 1 to 5 days, the pH value of the fermented bacterial liquid is kept between 3.5 and 4, and a fermented bacterial liquid 1 is obtained; adding 2.64% (w/w) of zymocyte liquid 1 into 20kg of fermentation substrate culture medium, controlling the water content of the final fermentation raw material to be kept at 60% by adding water, removing air, compacting and sealing, and carrying out anaerobic fermentation for 4 days at 28 ℃ in a dark place to obtain a grass-fungus fermentation composite microbial inoculum 1; inoculating 20% of the total mass of the composite bacterial grass fermentation inoculum 1 into 20kg of a new fermentation substrate culture medium, removing air, compacting, sealing, and performing anaerobic fermentation at 28 ℃ in a dark place for 4 days to obtain a composite bacterial grass fermentation inoculum 2; inoculating 20% of the total mass of the compound bacterial grass fermentation inoculum 2 into 20kg of a new fermentation substrate culture medium, removing air, compacting, sealing, and performing anaerobic fermentation at 28 ℃ in a dark place for 4 days to obtain a compound bacterial grass fermentation inoculum 3; inoculating 20% of the total mass of the composite bacterial grass fermentation inoculum 3 into 20kg of a new fermentation substrate culture medium, removing air, compacting, sealing, and performing anaerobic fermentation at 28 ℃ in a dark place for 4 days to obtain a composite bacterial grass fermentation inoculum 4.
Wherein the total viable bacteria concentration in the zymophyte liquid 1 is 1 multiplied by 108cfu/g, wherein the number ratio of the lactobacillus plantarum, the lactobacillus brevis, the lactobacillus japonicas, the lactobacillus paracasei, the han-jiu yeast abnormal, the lactobacillus buchneri, the bacillus subtilis and the coprinus pratense is 43: 35: 14: 6: 5: 4: 1.
Wherein the total viable bacteria concentration in the composite bacterial agent 4 for fermenting the grass is 2 multiplied by 109cfu/g. Make itExtracting the microbial DNA in the Juncao fermentation complex microbial inoculum 4 by using the kit, performing high-throughput sequencing on the extracted DNA, and calculating the abundance of microbial flora. The abundance of the microbial flora was calculated as follows: after high-throughput sequencing, splicing the obtained sequencing sequences, obtaining clean high-quality sequences after quality control, and then flattening all samples, wherein the abundance of the microorganisms of a certain species is the proportion of the sequence number of the species to the total sequence number of the microorganisms of the samples. As a result, the composite bacterial agent 4 for grass fermentation mainly contains the following beneficial bacteria in relative abundance: pantoea genus (Pantoea51.57%), Lactobacillus (L.) genusLactobacillus21.76%), Pediococcus (Pediococcus0.01%), enterococcus (E), and B), (E), (Enterococcus0.02%), Derlichia zeylanica (R) ((R)Naganishia1.68%), Pichia pastoris (Meyerozyma，5.31%）。
Example 3
Harvesting fresh Jujun grass with a growth period of 2 months, respectively drying the harvested fresh Jujun grass, wheat bran and corn flour at 50 ℃ for 12 h, and crushing into 20-mesh fine powder for later use; uniformly mixing 80 parts of megatherium powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; adding 2.5% (w/w) of the grass-fungus fermentation complex microbial inoculum 4 into the fermentation raw materials, adding water to control the water content of the final fermentation raw materials to be kept at 50%, removing air, compacting and sealing, and performing anaerobic fermentation at 30 ℃ in a dark place for 9 days to obtain the pennisetum sinese roxb fermented feed.
Harvesting fresh oasis I pasture with a growth period of 2 months, drying the harvested fresh oasis I pasture, wheat bran and corn flour at 50 ℃ for 12 hours respectively, and crushing into 20 meshes of fine powder for later use; uniformly mixing 80 parts of oasis I pasture powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; adding 2.5% (w/w) of a composite bacterial and grass fermentation inoculant 4 into the fermentation raw materials, adding water to control the water content of the final fermentation raw materials to be kept at 50%, removing air, compacting, sealing, and performing anaerobic fermentation at 30 ℃ in a dark place for 9 days to obtain the oasis I forage grass fermentation feed.
Example 4
Harvesting fresh Jujun grass with a growth period of 2 months, respectively drying the harvested fresh Jujun grass, wheat bran and corn flour at 50 ℃ for 12 h, and crushing into 20-mesh fine powder for later use; uniformly mixing 80 parts of megatherium powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; adding 1.5% (w/w) of the composite bacterial agent 4 into the fermentation raw material, adding water to control the water content of the final fermentation raw material to be 60%, removing air, compacting, sealing, and performing anaerobic fermentation at 35 ℃ in a dark place for 9 days to obtain the Jujun grass fermented feed.
Harvesting fresh oasis I pasture with a growth period of 2 months, drying the harvested fresh oasis I pasture, wheat bran and corn flour at 50 ℃ for 12 hours respectively, and crushing into 20 meshes of fine powder for later use; uniformly mixing 80 parts of oasis I pasture powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; adding 4.5% (w/w) of the composite bacterial agent 4 into the fermentation raw materials, adding water to control the water content of the final fermentation raw materials to be kept at 60%, removing air, compacting and sealing, and performing anaerobic fermentation at 35 ℃ in a dark place for 9 days to obtain the grassy fermented feed for oasis I.
Example 5
Harvesting fresh Jujun grass with a growth period of 2 months, respectively drying the harvested fresh Jujun grass, wheat bran and corn flour at 50 ℃ for 12 h, and crushing into 20-mesh fine powder for later use; uniformly mixing 80 parts of pennisetum hydridum powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; adding 3.5% (w/w) of the grass-fungus fermentation complex microbial inoculum 4 into the fermentation raw materials, adding water to control the water content of the final fermentation raw materials to be maintained at 70%, removing air, compacting and sealing, and carrying out anaerobic fermentation at 30 ℃ in a dark place for 9 days to obtain the pennisetum sinese roxb fermented feed.
Harvesting fresh oasis I pasture with a growth period of 2 months, drying the harvested fresh oasis I pasture, wheat bran and corn flour at 50 ℃ for 12 hours respectively, and crushing into 20 meshes of fine powder for later use; uniformly mixing 80 parts of oasis I pasture powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; adding 3.5% (w/w) of the composite bacterial agent 4 for fermenting the grass, adding water to control the water content of the final fermentation raw material to be 70%, removing air, compacting, sealing, and performing anaerobic fermentation at 30 ℃ in a dark place for 9 days to obtain the forage grass fermented feed of oasis I.
Example 6
Harvesting fresh Jujun grass with a growth period of 2 months, respectively drying the harvested fresh Jujun grass, wheat bran and corn flour at 50 ℃ for 12 h, and crushing into 20-mesh fine powder for later use; uniformly mixing 80 parts of megatherium powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; adding 2.5% (w/w) of the white rot fungi crude enzyme solution 2 and 2.5% (w/w) of the grass-fungus fermentation composite microbial inoculum 4 into the fermentation raw materials, adding water to control the water content of the final fermentation raw materials to be kept at 50%, removing air, compacting, sealing, and carrying out anaerobic fermentation at 30 ℃ in a dark place for 9 days to obtain the pennisetum sinese fermented feed.
Harvesting fresh oasis I pasture with a growth period of 2 months, drying the harvested fresh oasis I pasture, wheat bran and corn flour at 50 ℃ for 12 hours respectively, and crushing into 20 meshes of fine powder for later use; uniformly mixing 80 parts of oasis I pasture powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; adding 2.5% (w/w) of the white rot fungi crude enzyme liquid 2 and 2.5% (w/w) of the composite bacterial agent 4 for fermenting the grass and fungus into the fermentation raw materials, adding water to control the water content of the final fermentation raw materials to be kept at 50%, removing air, compacting, sealing, and carrying out anaerobic fermentation at 30 ℃ in a dark place for 9 days to obtain the forage grass fermented feed for oasis I.
Example 7
Harvesting fresh Jujun grass with a growth period of 2 months, respectively drying the harvested fresh Jujun grass, wheat bran and corn flour at 50 ℃ for 12 h, and crushing into 20-mesh fine powder for later use; uniformly mixing 80 parts of megatherium powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; adding 2.5% (w/w) of the white rot fungus crude enzyme solution 1 and 2.5% (w/w) of the grass-fungus fermentation composite microbial inoculum 4 into the fermentation raw materials, adding water to control the water content of the final fermentation raw materials to be kept at 50%, removing air, compacting, sealing, and carrying out anaerobic fermentation at 30 ℃ in a dark place for 9 days to obtain the pennisetum sinese fermented feed.
Harvesting fresh oasis I pasture with a growth period of 2 months, drying the harvested fresh oasis I pasture, wheat bran and corn flour at 50 ℃ for 12 hours respectively, and crushing into 20 meshes of fine powder for later use; uniformly mixing 80 parts of oasis I pasture powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; adding 2.5% (w/w) of the white rot fungi crude enzyme solution 1 and 2.5% (w/w) of the composite bacterial agent 4 for fermenting the grass and fungus into the fermentation raw materials, adding water to control the water content of the final fermentation raw materials to be kept at 50%, removing air, compacting, sealing, and carrying out anaerobic fermentation at 30 ℃ in a dark place for 9 days to obtain the forage grass fermented feed for oasis I.
Comparative example 1
Harvesting fresh Jujun grass with a growth period of 2 months, respectively drying the harvested fresh Jujun grass, wheat bran and corn flour at 50 ℃ for 12 h, and crushing into 20-mesh fine powder for later use; uniformly mixing 80 parts of megatherium powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; controlling the water content of the final fermentation raw material to be kept at 50% by adding water, removing air, compacting and sealing, and carrying out anaerobic fermentation at 30 ℃ in a dark place for 9 days to obtain the pennisetum sinese roxb fermented feed.
Harvesting fresh oasis I pasture with a growth period of 2 months, drying the harvested fresh oasis I pasture, wheat bran and corn flour at 50 ℃ for 12 hours respectively, and crushing into 20 meshes of fine powder for later use; uniformly mixing 80 parts of oasis I pasture powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material; controlling the water content of the final fermentation raw material to be kept at 50% by adding water, removing air, compacting and sealing, and carrying out anaerobic fermentation for 9 days at 30 ℃ in a dark place to obtain the oasis I pasture fermentation feed.
The fermented feeds obtained in the above examples and comparative examples were evaluated for quality by the method of "silage quality evaluation Standard" of the Ministry of agriculture, and were 15 points in color, 15 points in smell, and 10 points in texture. Wherein the superior is 40 to 30 minutes, the good is 30 to 20 minutes, generally 20 to 10 minutes, and the difference is less than 10 minutes. As can be seen from the results in tables 1 and 2, the sensory evaluation results of the fermented feeds obtained in the examples are all good and above, and the total sensory evaluation scores of the fermented feeds obtained in examples 3 to 7 are higher than that of the fermented feed obtained in comparative example 1, wherein the total sensory evaluation scores of the pennisetum sinese fermented feed and the grassy fermented feed for oasis number one obtained in examples 3, 6 and 7 are all superior and have light yellow green or light yellow green color, and the fermented feeds have tart aroma and green grass sweet aroma, are slightly moist in texture, and are not easy to be scattered when being agglomerated by soft hands.
TABLE 1 sensory evaluation results of Jujun grass fermented feed
TABLE 2 sensory evaluation results of oasis grass fermented feed
The chemical components of the fermented feed obtained in the above examples were measured. The Dry Matter (DM) and the Crude Protein (CP) are respectively determined according to GB/T6435-2014 and GB/T6432-2018. Determination of Neutral Detergent Fiber (NDF), Acid Detergent Fiber (ADF), Acid Detergent Lignin (ADL) and Acid Insoluble Ash (AIA), and calculation reference of cellulose and hemicellulose content [ Songhong. mowing on yield and nutritive value of Medicago sativa and Ormosia herbacea [ D]A method of Yangshi academic paper, Yangshi, North-west agriculture and forestry Science and technology university, 2016, reference for calculating relative feed value (RFQ) [ ROHWEDER D A, BARNES R F, JORGENSEN N. deployed had grazing standards based on laboratories for evaluating quality, Journal of Animal Science,1978,47:747-]The title of tropical crops, 2013,34(02): 377-. Ammoniacal Nitrogen (NH)3-N) content was determined using a phenol-sodium hypochlorite colorimetric method, with particular reference to [ BRODERICK G A, KANG J H. Automated multiple amino determination of ammonia and total amino acids in a crude fluid and in a visual medium [ J ]]Journal of Dairy Science,1980,63(1). Volume 63:64-75. As can be seen from tables 3 and 4, in the results of various index detections, in comparison with comparative example 1, the contents of pH, NDF, ADF, cellulose and hemicellulose in examples 3 to 7 are all reduced, so that the fiber structure of the fermented feed can be better changed,the digestibility and palatability of the fermented feed are improved, and more nutrient substrates are provided for the lactic acid bacteria. During the fermentation process of the feed, protein and amino acid are degraded to generate ammoniacal nitrogen, so that the quality of silage can be seen from the high content and the low content of ammoniacal nitrogen, and the lower the ammoniacal nitrogen value, the higher the ratio of ammoniacal nitrogen to total nitrogen is, the more the decomposition of protein and amino acid is; it can be seen from the table that the value of ammonia nitrogen in the non-bacteria-added group of the comparative example is lower, probably because the silage does not start to enter the fermentation period, and the non-decomposition nutrient does not produce a large amount of ammonia nitrogen, and the value is lower, while the value of other groups is in a controllable range and the like. In conclusion, the result of the Jujun grass fermented feed is the best in example 7, and the effect of the fermented feed of oasis I is the best in example 6.
TABLE 3 chemical composition index detection results of Jujun grass fermented feed
Table 4 chemical composition index detection results of oasis a pasture fermented feed
The above description is only a preferred embodiment of the present invention, and not intended to limit the present invention, and the scope of the present invention is defined by the appended claims, and all changes that are equivalent to the structural changes made by the present specification shall be embraced within the scope of the present invention.
Claims (8)
1. A method for preparing a compound flora feed by utilizing Juncao is characterized by comprising the following steps of: the composite flora feed is prepared by taking mushroom grass as a main material and corn flour and wheat bran as auxiliary materials, adding a white rot fungus crude enzyme solution to degrade cellulose, hemicellulose and lignin, and simultaneously adding a mushroom grass fermentation composite microbial inoculum for mixed fermentation.
2. The method of claim 1, wherein: the method specifically comprises the following steps:
(1) harvesting fresh grass in the growth period of 2 months;
(2) respectively drying the harvested fresh Jujun grass, wheat bran and corn flour at 50 ℃ for 12 h, and crushing into 20-mesh fine powder for later use;
(3) uniformly mixing 80 parts of megatherium powder, 10 parts of wheat bran powder and 10 parts of corn powder to obtain a fermentation raw material;
(4) adding 0.05% (w/w) of white rot fungus crude enzyme liquid and 1.5-4.5% (w/w) of a grass-fungus fermentation composite microbial inoculum into a fermentation raw material, adding water to control the water content of the final fermentation raw material to be kept at 50-70%, removing air, compacting, sealing, and carrying out anaerobic fermentation at 25-30 ℃ in a dark place for 9 days to obtain the composite flora feed.
3. The method for preparing a composite flora feed by using juncao as claimed in claim 1, wherein the method comprises the following steps: the Juncao is selected from Jujuncao, oasis No. 1, oasis No. 2, oasis No. 3 and oasis No. 5.
4. The method of claim 1, wherein: the preparation method of the white rot fungi crude enzyme solution comprises the following steps: inoculating single colony of white rot fungi into a PDA solid culture medium, performing static culture at 28 ℃ for 5 days, taking 4 hypha blocks with the diameter of 5mm, transferring the hypha blocks into 200 mL of PDB liquid culture medium, performing culture at 28 ℃ and 160rpm/min for 5 days, forming hypha balls with the diameter of 1-3mm in the culture solution by utilizing the self-flocculation characteristic of the white rot fungi, and filtering the culture solution containing the hypha balls by using 4 layers of sterile gauze to obtain filtrate, namely the crude white rot fungi enzyme solution.
5. The method of claim 1, wherein: the preparation method of the composite bacterial agent for fermenting the grass comprises the following steps: uniformly mixing 90g of brown sugar and 1kg of sterile water, decocting, cooling to 30 ℃, adding 2.5g of lactobacillus plantarum strain, 2g of lactobacillus brevis strain, 1.2g of lactobacillus sakakii strain, 1g of lactobacillus paracasei strain, 1g of abnormal vekhem yeast strain, 0.8g of lactobacillus buchneri strain, 0.25g of bacillus subtilis strain and 0.25g of coprinus pratense strain, uniformly stirring, putting into a closed fermentation container, keeping the fermentation temperature at 30 ℃, and continuously fermenting for 1-5 days to obtain a fermented bacterial liquid; adding 2.64% (w/w) of zymocyte liquid into 20kg of fermentation substrate culture medium, adding water to control the water content of the final fermentation raw material to be kept at 60%, removing air, compacting and sealing, carrying out anaerobic fermentation at 28 ℃ in a dark place, taking 4 days as a fermentation period, and carrying out continuous passage for four times to obtain the composite bacterial agent for fermenting the bacteria and the grass.
6. The method of claim 3, wherein: the fermentation substrate culture medium is prepared by mixing 16kg of megaterium powder, 2kg of wheat bran powder and 2kg of corn powder.
7. The method of claim 3, wherein: the total viable bacteria concentration in the zymophyte liquid is 1 multiplied by 108cfu/g, wherein the number ratio of the lactobacillus plantarum, the lactobacillus brevis, the lactobacillus japonicas, the lactobacillus paracasei, the han-jiu yeast abnormal, the lactobacillus buchneri, the bacillus subtilis and the coprinus pratense is 43: 35: 14: 6: 5: 4: 1.
8. The method of claim 3, wherein: the total viable bacteria concentration in the composite bacterial agent for fermenting the grass is 2 multiplied by 109cfu/g, containing mainly the following relative abundances of the genus probiotic: pantoea genus (Pantoea51.57%), Lactobacillus (L.) genusLactobacillus21.76%), Pediococcus (Pediococcus0.01%), enterococcus (E), and B), (E), (Enterococcus0.02%), Derlichia zeylanica (R) ((R)Naganishia1.68%), Pichia pastoris (Meyerozyma，5.31%）。
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| CN115462446A (en) * | 2022-08-11 | 2022-12-13 | 仪陇县宏兴畜禽养殖农民专业合作社 | Wild grass composite silage, preparation method thereof and poultry feeding feed |
| CN115530325A (en) * | 2022-10-20 | 2022-12-30 | 江南大学 | Chickpea powder rich in gamma-aminobutyric acid and application thereof |
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|---|---|---|---|---|
| CN115449501A (en) * | 2022-07-25 | 2022-12-09 | 福建华闽晟业生物科技有限公司 | Application of Jujun grass juice as non-pathogenic fusarium oxysporum fermentation culture medium |
| CN115462446A (en) * | 2022-08-11 | 2022-12-13 | 仪陇县宏兴畜禽养殖农民专业合作社 | Wild grass composite silage, preparation method thereof and poultry feeding feed |
| CN115530325A (en) * | 2022-10-20 | 2022-12-30 | 江南大学 | Chickpea powder rich in gamma-aminobutyric acid and application thereof |
| CN115530325B (en) * | 2022-10-20 | 2024-02-27 | 江南大学 | Chickpea flour rich in gamma-aminobutyric acid and application thereof |
| CN116831224A (en) * | 2023-07-26 | 2023-10-03 | 楚雄师范学院 | Preparation method of fermented pennisetum sinese feed |
| CN116831224B (en) * | 2023-07-26 | 2024-01-26 | 楚雄师范学院 | Preparation method of fermented pennisetum sinese feed |
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