CN112262916B - Complex microbial inoculum fermentation product and preparation method and application thereof - Google Patents
Complex microbial inoculum fermentation product and preparation method and application thereof Download PDFInfo
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- 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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
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- 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
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
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- A23K—FODDER
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
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- 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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- A—HUMAN NECESSITIES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
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- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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- A—HUMAN NECESSITIES
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- A23K20/163—Sugars; Polysaccharides
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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
- A23K40/00—Shaping or working-up of animal feeding-stuffs
- A23K40/10—Shaping or working-up of animal feeding-stuffs by agglomeration; by granulation, e.g. making powders
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- 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
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
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Abstract
The invention discloses a composite microbial inoculum fermentation product and a preparation method and application thereof. 46, mixing the soybean meal and the bran, adding water for pre-dilution to prepare a solid culture medium, then adding the antibacterial peptide, the glucose oxidase, the glucose and the water for fully mixing, and fermenting for 12-24 hours to obtain a fermented material for later use. Adding lactobacillus plantarum, saccharomyces cerevisiae, bacillus subtilis and cellulase into the fermentation material, adding water again for humidification, sealing and storing, standing and fermenting for 48-72 h at the constant temperature of 35 ℃ to obtain the composite microbial inoculum fermentation product. The composite microbial inoculum fermentation product prepared by the method has higher biological activity, the functional animal feed prepared by mixing the composite microbial inoculum fermentation product and the puffed corn can improve the nutritional effect of the feed, reduce the incidence rate of animal diseases, effectively replace part of antibiotics, and has simple, easy, green and environment-friendly process and wider application prospect in the feed industry and the breeding industry.
Description
Technical Field
The invention belongs to the technical field of biological fermentation, and particularly relates to a composite microbial inoculum fermentation product, and a preparation method and application thereof.
Background
In animal husbandry production, the application of antibiotics can improve the utilization efficiency and economic benefit of the feed. However, the abuse of antibiotics in the feed causes pathogenic microorganisms of livestock and poultry to generate drug resistance and superinfection; the livestock and poultry products have drug residues, the quality and the safety of the products are seriously affected, and the health of human beings is threatened, so that the search for high-efficiency and green antibiotic substitute products becomes a research hotspot, and since the last 90 th century, many researchers at home and abroad begin to search for antibiotic substitutes. In addition, the conventional raw material resources are in short supply, the utilization is relatively limited, and the screening of the non-conventional feed raw materials is also a necessary trend. The fermented feed utilizes the growth, the propagation and the metabolism of microorganisms in raw materials to accumulate useful thalli, enzymes and metabolites to produce, process and modulate the feed, is expected to become a substitute of antibiotics to be applied to the field of livestock raising, and can also improve the utilization rate of the raw materials of the feed.
The soybean meal and the bran are adopted as raw materials for combined fermentation, so that anti-nutritional factors and oligosaccharides can be effectively removed, macromolecular protein is degraded into peptide substances which are easy to digest and utilize, the digestion and utilization rate of the peptide substances is improved, and fermentation can generate fermentation products with various flavors and organic acid; in addition, the bran can also degrade mycotoxin after fermentation, and the palatability of the feed is improved, so that the feed intake of animals is improved. However, in the combined fermentation process, the bran has high content of mixed bacteria, and the bran is considered to be too high in cost and not subjected to high-temperature treatment, so that the pollution is serious in the fermentation process, and the fermentation effect is influenced by more mycotoxins, particularly vomitoxin, in the fermentation process. If high-temperature treatment is simply added to the fermentation raw materials, the cost is high, and the bioactive substances in the fermentation raw materials are inactivated when meeting high temperature, so that the nutrient components of the feed are seriously damaged. The advent of antimicrobial peptides has allowed the above problems to be partially solved.
Antimicrobial peptides (A), (B), (Cantimicrobial peptides，AMPs) Is an important component of the natural defense system of most organisms against invading pathogens, has a unique antibacterial action mechanism, has quick bactericidal action and is not easy to cause drug resistance of bacteria, and can be used for killing pathogens singly or in combination with antibiotics. In contrast to traditional antibiotics, most antimicrobial peptides exhibit multiple biological activities, acting primarily by affecting the cytoplasmic membrane. Therefore, bacteria must change the composition and structure of their membranes to develop resistance to antimicrobial peptides, which can cause serious damage to the bacteria themselves. The outer layer of the bacterial cell membrane is rich in anionic phospholipids, and the zwitterionic lipids of the mammalian cell membrane are rich, which is the main reason why the antibacterial peptide can resist bacteria and does not damage mammalian cells. Moreover, studies have shown that the antimicrobial peptides have immunomodulatory functions, such as chemotaxis, wound healing, endotoxin resistance, etc.
In intensive research over the past decades, various theoretical hypotheses have been proposed to explain the process of antimicrobial peptides to inhibit or kill microorganisms. However, the antibacterial mechanism of antibacterial peptide is only studied for individual antibacterial peptides, so that at present, there is no hypothesis which can cover the action mechanism of all kinds of antibacterial peptides, and it is uncertain which hypothesis is closer to the real situation. In addition, the natural antibacterial peptide has limited sources, complex extraction process and high cost, and the chemical synthesis has high cost, difficult batch production and the like. In addition, the natural antibacterial peptide is processed and modified after being synthesized in an organism and has a certain spatial structure, and the antibacterial activity of the natural antibacterial peptide is closely related to the spatial structure of peptide molecules. Although the primary structure of the in vitro synthesized antibacterial peptide is consistent with that of the natural antibacterial peptide, the spatial structure of the in vitro synthesized antibacterial peptide is different from that of the natural antibacterial peptide, so that the activity is poor.
Based on the analysis, how to ensure that the antibacterial peptide is applied to the production and application of animal feed, reduce harmful microorganisms in products and promote the growth of beneficial microorganisms is urgently needed in the industry at present.
Disclosure of Invention
In view of the defects, the invention solves the defect that the product is easily polluted by microorganisms in the production process by adding the antibacterial peptide into the jointly fermented animal feed, and simultaneously solves the technical problem of low feed nutrition activity content caused by high-cost high-temperature sterilization in the traditional feed processing process. In addition, the invention also unexpectedly discovers that under the condition of using the formula of the antibacterial peptide and combining other biological agents and processes, compared with the traditional method of simply adding the antibacterial peptide, the prepared composite microbial inoculum fermentation product has the advantages that the content of lactic acid bacteria is obviously improved, mycotoxin is effectively controlled, the beneficial bacteria number and peptide substances in the fermentation material are obviously improved, the comprehensive use effect of the product is improved, and the feed is safer. In order to solve the technical problems, the invention adopts the following technical scheme:
a preparation method of a composite microbial inoculum fermentation product comprises the following steps:
(1) and (3) an antibacterial stage: selecting high-quality 46 soybean meal, mixing with bran, adding water for pre-dilution to prepare a solid culture medium, then adding antibacterial peptide, glucose oxidase, glucose and water for fully mixing, and then fermenting to obtain a fermented material for later use;
(2) solid state fermentation stage: adding 1/10 times of composite strain and 1/60 times of cellulase into the fermentation material obtained in the antibacterial stage, adding water again for humidification, sealing and storing, and standing at constant temperature for fermentation to obtain the composite microbial inoculum fermentation product.
Further, the mass ratio of the high-quality 46 soybean meal to the bran in the step (1) is 6:4 or 8: 2.
Further, the mass ratio of the high-quality 46 soybean meal to the bran is 7: 3.
Further, the addition percentages of the antibacterial peptide, the glucose oxidase and the glucose in the step (1) respectively account for 0.1%, 10% and 20% -100% of the solid culture medium, and preferably account for 0.1%, 10% and 50%.
Further, the fermentation condition temperature in the step (1) is 30-35 ℃, and the fermentation time is 12-24 h.
Further, the composite bacterial strain in the step (2) is prepared by mixing lactobacillus plantarum, saccharomyces cerevisiae and bacillus subtilis according to the mass ratio of 1:1: 1.
Further, the constant-temperature fermentation conditions in the step (2) are as follows: the temperature is 30-35 ℃; the time is 48-72 h.
Further, the water content of the fermentation product of the complex microbial inoculum in the step (2) is 35-40%.
The invention also discloses a composite microbial inoculum fermentation product prepared by any preparation method.
The invention also discloses application of the composite microbial inoculum fermentation product in preparing functional animal feed.
Further, the application comprises:
and (3) granulating the composite microbial inoculum fermentation product and the puffed corn in a cold chain to obtain the functional animal feed.
Further, the mass ratio of the composite microbial inoculum fermentation product to the puffed corn is 3:7 or 2.5: 7.5.
The invention has the beneficial effects that:
1. compared with the traditional method of simply adding the composite bacteria, the biological activity of the prepared composite bacteria fermentation product is higher, the crude protein content is 54.5%, the acid soluble protein (also called small peptide) content is 15.48%, and the content of the lactobacillus plantarum is 7.80 multiplied by 109cfu/g, the bacteriostatic potency of the antibacterial peptide reaches 3.0 multiplied by 104cfu/g~9×105cfu/g, so that mycotoxin in the fermentation process of the product is effectively controlled, the beneficial bacteria number and peptide substances in the fermentation material are obviously improved, the comprehensive use effect of the product is improved, and the feed is safer.
2. The composite microbial inoculum fermentation product prepared by the process is applied to the production and processing of the soybean meal, not only can improve the nutritional efficacy of the soybean meal and reduce the occurrence probability of animal diarrhea, but also can replace part of antibiotics, and has simple and easy process, environmental protection and wide application prospect in the feed industry and the breeding industry.
3. The compound microbial inoculum fermentation product prepared by the invention and the puffed corn are granulated in a cold chain, so that the damage of microorganisms and metabolites such as digestive enzymes, unknown growth factors, vitamins and the like generated by the microorganisms in the fermentation process by high temperature can be avoided, stable internal conditions are provided for the production of animal feed, and the quality of the prepared animal feed is effectively guaranteed.
Detailed Description
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto.
The strains used in the examples of the present invention were conventional commercially available strains. The lactobacillus plantarum and the saccharomyces cerevisiae are purchased from microbial technology limited company in Hubei blue valley, and the production batches are respectively as follows: LGZW2020042801, LGZW 20200428802. The bacillus subtilis is purchased from Shandong root source biological technology limited company, and the production batches are as follows: GY 20200524108. The antibacterial peptide is purchased from Shengxi Ma biotechnology Limited company in Shenzhen, and the production batches are as follows: 20200514003.
example 1
Composite microbial inoculum fermentation product
(1) And (3) an antibacterial stage: selecting 70kg of high-quality 46 soybean meal, mixing with 30kg of dry bran (middle bran), adding water for pre-dilution to prepare a solid culture medium, then adding 0.1kg of antibacterial peptide, 10kg of glucose oxidase and 50kg of glucose, fully mixing with water, and then fermenting at 32 ℃ for 18 hours to obtain a fermented material for later use;
(2) solid state fermentation stage: 1/10 times of composite strain and 1/60 times of cellulase are added into the fermentation material obtained in the antibacterial stage, and the composite strain is formed by mixing lactobacillus plantarum, saccharomyces cerevisiae and bacillus subtilis according to the mass ratio of 1:1: 1; adding water again for humidification, sealing and storing, standing and fermenting for 60h at the constant temperature of 32 ℃ to obtain a composite microbial inoculum fermentation product, wherein the water content of the composite microbial inoculum fermentation product is 37.5%.
Example 2
Composite microbial inoculum fermentation product
(1) And (3) an antibacterial stage: selecting 60kg of high-quality 46 soybean meal, mixing with 40kg of dry bran (middle bran), adding water for pre-dilution to prepare a solid culture medium, then adding 0.1kg of antibacterial peptide, 10kg of glucose oxidase, 20kg of glucose and water for fully mixing, and then fermenting at 30 ℃ for 24 hours to obtain a fermented material for later use;
(2) solid state fermentation stage: 1/10 times of composite strain and 1/60 times of cellulase are added into the fermentation material obtained in the antibacterial stage, and the composite strain is formed by mixing lactobacillus plantarum, saccharomyces cerevisiae and bacillus subtilis according to the mass ratio of 1:1: 1; adding water again for humidification, sealing and storing, standing and fermenting for 72 hours at the constant temperature of 30 ℃ to obtain a composite microbial inoculum fermentation product, wherein the water content of the composite microbial inoculum fermentation product is 35%.
Example 3
Composite microbial inoculum fermentation product
(1) And (3) an antibacterial stage: selecting 80kg of high-quality 46 soybean meal, mixing with 20kg of dry bran (middle bran), adding water for pre-dilution to prepare a solid culture medium, then adding 0.1kg of antibacterial peptide, 10kg of glucose oxidase and 100kg of glucose, fully mixing with water, and then fermenting at 35 ℃ for 12 hours to obtain a fermented material for later use;
(2) solid state fermentation stage: 1/10 times of composite strain and 1/60 times of cellulase are added into the fermentation material obtained in the antibacterial stage, and the composite strain is formed by mixing lactobacillus plantarum, saccharomyces cerevisiae and bacillus subtilis according to the mass ratio of 1:1: 1; adding water again for humidification, sealing and storing, standing and fermenting for 72h at the constant temperature of 30 ℃ to obtain a composite microbial inoculum fermentation product, wherein the water content of the composite microbial inoculum fermentation product is 40%.
Comparative example 1
In contrast to example 1, the use of antimicrobial peptides was eliminated
(1) And (3) an antibacterial stage: selecting 70kg of high-quality 46 soybean meal, mixing with 30kg of dry bran (medium bran), adding water for pre-dilution to prepare a solid culture medium, then adding 10kg of glucose oxidase, 50kg of glucose and water for fully mixing, and then fermenting at 32 ℃ for 18 hours to obtain a fermented material for later use;
(2) solid state fermentation stage: 1/10 times of composite strain and 1/60 times of cellulase are added into the fermentation material obtained in the antibacterial stage, and the composite strain is formed by mixing lactobacillus plantarum, saccharomyces cerevisiae and bacillus subtilis according to the mass ratio of 1:1: 1; adding water again for humidification, sealing, storing, standing at constant temperature of 32 deg.C, and fermenting for 60 hr to obtain fermented product.
Comparative example 2
In contrast to example 1, the use of Bacillus subtilis was eliminated
(1) And (3) an antibacterial stage: selecting 70kg of high-quality 46 soybean meal, mixing with 30kg of dry bran (middle bran), adding water for pre-dilution to prepare a solid culture medium, then adding 0.1kg of antibacterial peptide, 10kg of glucose oxidase and 50kg of glucose, fully mixing with water, and then fermenting at 32 ℃ for 18 hours to obtain a fermented material for later use;
(2) solid state fermentation stage: 1/10 times of composite strain and 1/60 times of cellulase are added into the fermentation material obtained in the antibacterial stage, and the composite strain is formed by mixing lactobacillus plantarum and saccharomyces cerevisiae according to the mass ratio of 1: 1; adding water again for humidification, sealing, storing, standing at constant temperature of 32 deg.C, and fermenting for 60 hr to obtain fermented product.
Comparative example 3
In contrast to example 1, the use of various enzymes in two stages (glucose oxidase in the first stage and cellulase in the second stage) was eliminated
(1) And (3) an antibacterial stage: selecting 70kg of high-quality 46 soybean meal, mixing with 30kg of dry bran (middle bran), adding water for pre-dilution to prepare a solid culture medium, then adding 0.1kg of antibacterial peptide, 50kg of glucose and water for fully mixing, and then fermenting at 32 ℃ for 18 hours to obtain a fermented material for later use;
(2) solid state fermentation stage: adding 1/10 times of composite strains into the fermentation material obtained in the antibacterial stage, wherein the composite strains are formed by mixing lactobacillus plantarum, saccharomyces cerevisiae and bacillus subtilis according to the mass ratio of 1:1: 1; adding water again for humidification, sealing, storing, standing at constant temperature of 32 deg.C, and fermenting for 60 hr to obtain fermented product.
Comparative example 4
In contrast to example 1, the use of first stage glucose oxidase was eliminated
(1) And (3) an antibacterial stage: selecting 70kg of high-quality 46 soybean meal, mixing with 30kg of dry bran (middle bran), adding water for pre-dilution to prepare a solid culture medium, then adding 0.1kg of antibacterial peptide, 50kg of glucose and water for fully mixing, and then fermenting at 32 ℃ for 18 hours to obtain a fermented material for later use;
(2) solid state fermentation stage: 1/10 times of composite strain and 1/60 times of cellulase are added into the fermentation material obtained in the antibacterial stage, and the composite strain is formed by mixing lactobacillus plantarum, saccharomyces cerevisiae and bacillus subtilis according to the mass ratio of 1:1: 1; adding water again for humidification, sealing, storing, standing at constant temperature of 32 deg.C, and fermenting for 60 hr to obtain fermented product.
Comparative example 5
Compared with example 1, the elimination of the second stage eliminates the use of cellulase
(1) And (3) an antibacterial stage: selecting 70kg of high-quality 46 soybean meal, mixing with 30kg of dry bran (middle bran), adding water for pre-dilution to prepare a solid culture medium, then adding 0.1kg of antibacterial peptide, 10kg of glucose oxidase and 50kg of glucose, fully mixing with water, and then fermenting at 32 ℃ for 18 hours to obtain a fermented material for later use;
(2) solid state fermentation stage: adding 1/10 times of composite strains into the fermentation material obtained in the antibacterial stage, wherein the composite strains are formed by mixing lactobacillus plantarum, saccharomyces cerevisiae and bacillus subtilis according to the mass ratio of 1:1: 1; adding water again for humidification, sealing, storing, standing at constant temperature of 32 deg.C, and fermenting for 60 hr to obtain fermented product.
Test example 1
The number of viable bacteria in the composite microbial inoculum fermentation products of example 1 and comparative examples 1-5 is determined by adopting a GBT 26428-2010 method; determining the total number of moulds in the feed by a GB/T13092-2006 method; measuring the content of acid soluble protein by using a QB/T2653-2004 method; determining the bacteriostatic potency of the antibacterial peptide by an oxford cup method; and (3) determining the content of the crude protein by adopting a GBT 6432-2018 method. The results are shown in Table 1.
TABLE 1 results of measurement of Properties of fermentation products of each group
From the results shown in Table 1, the fermentation products of the group of example 1 showed the highest biological activity and the best antibacterial effect. Wherein comparative example 1 compared to example 1, the use of antimicrobial peptide was eliminated, and as a result, it was found that: because of the action of glucose oxidase and glucose acidogenesis in the first stage antibacterial stage, the synergistic effect of the antibacterial peptide is lacked; the second stage of the fermentation is relatively full due to the action of the bacterial enzyme, the antibacterial peptide titer generated by the fermentation product is higher, the mold content is lower, but the fermentation effect is lower than that of the example 1. Comparative example 2 compared to example 1, the use of bacillus subtilis was eliminated and as a result: the synergistic effect of glucose oxidase and glucose acidogenesis in the first stage of antibacterial phase and the antibacterial peptide; in the second stage, the enzymolysis of the bacillus subtilis on bran and soybean meal is lacked, so that the fermentation product has higher toxin and higher mould content, which is lower than the fermentation effect of the embodiment 1. Comparative example 3 compared to example 1, the use of various enzymes in two stages (glucose oxidase elimination in the first stage and cellulase elimination in the second stage) was eliminated, and as a result, it was found that: the bacteriostasis effect of the antibacterial peptide is relatively weakened, and the fermentation and enzymolysis effects of the compound bacteria make the content of the mould in the fermentation product and the antibacterial effect of the antibacterial peptide lower than those of the example 2. Comparative example 4 compared to example 1, the use of first stage glucose oxidase was eliminated and as a result: the antimicrobial peptide plays a role in the first stage of the antimicrobial phase, but lacks the synergistic effect of glucose oxidase, so that the antimicrobial inhibition effect is weakened, and the fermentation effect is relatively better than that of the example 1 due to the synergy in the later fermentation process. Comparative example 5 compared to example 1, the elimination of the second stage of the use of cellulase resulted in the finding that: the influence of cellulase on the fermentation effect is greater than that of glucose oxidase, and the fermentation effect is poor and lower than that of the embodiment 1. Therefore, the fermentation product prepared by the process formula can obtain the highest biological activity and antibacterial effect.
Test example 2
And respectively granulating the composite microbial inoculum fermentation products prepared in the example 1 and the comparative examples 1-5 with puffed corn in a cold chain manner to obtain the functional animal feed. Wherein the mass ratio of the fermentation product to the puffed corn is 3: 7. The biological indexes of the granulated fermentation products of each group were measured, and the specific results are shown in table 2.
TABLE 2 comparison of the biological indicators after granulation of the fermentation products of the various groups
According to the results shown in table 2, after the fermented product and the puffed corn are mixed and granulated according to the mass ratio of 3:7, the low-temperature granulation has small influence on the crude protein content, the acid soluble protein content, the antibacterial potency of the lactobacillus plantarum and the antibacterial peptide, and the prepared animal feed has the highest biological activity and the best antibacterial effect after the fermented product and the puffed corn of the group 1 are subjected to cold chain granulation.
Test example 3
The functional animal feed prepared in the example 1 and the comparative examples 1-5 is used for feeding animals, and meanwhile, the same type of animals fed by basic ration are selected as a control group, and the indexes of the animals in the example 1 group, the comparative examples 1-5 groups and the control group are compared.
1. Materials and methods
1.1 design of the experiment
210 pigs with consistent sources and age of 162 days and weight of about 80kg are randomly divided into seven groups, each group has 5 repetitions, and each repetition has 6 pigs. Test cycle 30d, a specific test design is shown in table 3.
TABLE 3 test design
1.2 test daily ration
The test material was produced in the company's test shop 1 week in advance using the company's existing base stock formulation.
1.3 Breeding management
And carrying out a feeding management test in a fattening house of a test pig farm in 7-8-2020-8-3 days, wherein the test time is 27 days. Fattening pigs with uniform sizes are selected, the daily ages of the piglets are 168-194 days, the piglets are randomly divided into seven groups, the initial weights are weighed and divided into groups, and different feeds are fed to the groups respectively. The pigs are fed for 2 times a day (8: 00, 16: 00) to ensure that the pigs can freely eat and drink water. The feeding, mental and fecal conditions, weather conditions, temperature, humidity, etc. of the pigs are observed and recorded at regular intervals (8: 30, 16: 30) every day, and daily records are made. If diseases or death occurs, the patients should be treated symptomatically or eliminated immediately according to actual conditions and recorded. During the test, the pigsty is kept clean and air is circulated, the pigsty is cleaned twice a day and disinfected 3 times a week (a plurality of disinfectants are used alternately).
2. Statistical index
And (4) counting initial weight, final weight, weekly feed intake, feed-meat ratio, diarrhea rate and diarrhea index, wherein the feed intake, the shape of excrement and the quantity of excrement are focused.
During the trial, pigs were observed at 7:00 am and 17:00 pm for fecal discharge each day, and the first diarrhea was recorded and scored.
Diarrhea rate = (diarrhea first/total number of heads) × 100%;
diarrhea index = (1 min. 1+2 min. 2+3 min. 3)/total diarrhea;
diarrhea index evaluation criteria:
1 minute: mild diarrhea, the shape of the excrement is close to the shape of normal excrement, and the excrement is slightly soft and formable;
and 2, dividing: moderate diarrhea, high water content of feces, and pasty feces, which can be accumulated but not shaped;
and 3, dividing: severe diarrhea: the excrement is in a water sample shape and is in a spraying shape when being discharged.
Third, test results
The test data were analyzed, and the specific results are shown in Table 4.
Table 4 test results of each group
As can be seen from Table 4, the group of example 1 had the highest daily gain, the highest feed intake, the lowest feed-meat ratio, the lowest diarrhea rate and the best growth performance. Therefore, the fermented feed with the concentration of 5% can improve the growth performance of the fattening pigs and reduce diarrhea. The fermented feed produced by the method disclosed by the patent is the fermented feed produced by the embodiment 1, namely, the feed has the highest daily gain and feed intake, the lowest feed-meat ratio, the highest growth performance, the lowest diarrhea rate and the like.
It should be noted that the technical effect of the present invention is the result of the mutual cooperation and interaction of the process steps and parameters, and is not the superposition of simple processes, and the effect produced by the organic combination of the processes far exceeds the superposition of the functions and effects of each single process, so the present invention has better advancement and practicability.
The above description is only one specific guiding embodiment of the present invention, but the design concept of the present invention is not limited thereto, and any insubstantial modification of the present invention using this concept shall fall within the scope of the invention.
Claims (6)
1. A preparation method of a composite microbial inoculum fermentation product comprises the following steps:
(1) preparing a fermentation material in an antibacterial stage:
46 soybean meal and bran in a mass ratio of 6:4 or 8:2 or 7:3 are selected and mixed, water is added for pre-dilution, a solid culture medium is prepared, then antibacterial peptide, glucose oxidase, glucose and water are added and fully mixed, and the addition percentage of the antibacterial peptide, the glucose oxidase and the glucose respectively accounts for 0.1%, 10% and 20% -100% of the solid culture medium; then fermenting for 12-24 h at 30-35 ℃ to obtain a fermented material for later use;
(2) preparing a composite microbial inoculum fermentation product in a solid state fermentation stage:
adding 1/10 times of composite strain and 1/60 times of cellulase into the fermentation material obtained in the antibacterial stage, adding water again for humidification, sealing and storing, and standing at constant temperature for fermentation to obtain a composite microbial inoculum fermentation product;
wherein the composite strain is prepared by mixing lactobacillus plantarum, saccharomyces cerevisiae and bacillus subtilis according to the mass ratio of 1:1: 1;
the water content of the prepared composite microbial inoculum fermentation product is 35-40%, the crude protein content is 54.5%, the acid soluble protein content is 15.48%, and the content of lactobacillus plantarum is 7.80 multiplied by 109cfu/g, the bacteriostatic potency of the antibacterial peptide is 3.0 multiplied by 104cfu/g~9×105cfu/g。
2. The preparation method of the composite microbial inoculum fermentation product according to claim 1, wherein the constant-temperature standing fermentation temperature is 30-35 ℃; the fermentation time is 48-72 h.
3. A complex microbial inoculum fermentation product prepared by the method for preparing the complex microbial inoculum fermentation product according to any one of claims 1 or 2.
4. The use of the complex microbial agent fermentation product according to claim 3 in the preparation of functional animal feed.
5. The use of claim 4, which comprises granulating the composite microbial agent fermentation product and the puffed corn cold chain to obtain the functional animal feed.
6. The use of claim 5, wherein the mass ratio of the composite microbial inoculum fermentation product to the puffed corn is 3:7 or 2.5: 7.5.
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