CN114107120A - Microbial agent for straw fermentation padding and application of microbial agent in healthy breeding of live pigs - Google Patents
Microbial agent for straw fermentation padding and application of microbial agent in healthy breeding of live pigs Download PDFInfo
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- 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
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K1/00—Housing animals; Equipment therefor
- A01K1/015—Floor coverings, e.g. bedding-down sheets ; Stable floors
- A01K1/0152—Litter
- A01K1/0155—Litter comprising organic material
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/14—Fungi; Culture media therefor
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- C12N1/18—Baker's yeast; Brewer's yeast
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Abstract
The invention discloses a microbial agent for straw fermentation padding and application thereof in healthy breeding of live pigs, wherein the microbial compound microbial agent is a compound flora preparation taking lactic acid bacteria, bacillus, saccharomycetes and the like as basic strains. According to the invention, conditions such as initial culture medium, fermentation temperature, fermentation pH, fermentation time, rotating speed and the like are selected and optimized through orthogonal design on three main types of lactic acid bacteria, bacillus and saccharomycetes in the flora preparation, an optimal high-density preparation system is finally established, the strains are independently fermented and then mixed according to respective proportions to prepare the composite microbial inoculum, the number of viable bacteria of the microbial inoculum meets the requirement of commercialization on the density of the microbial inoculum, the construction of the special flora preparation for raising pigs by using straw fermentation padding is completed, the preparation can be used for healthy culture of live pigs, the effects of zero discharge of feces and quasi zero discharge of odor are achieved, and the pollution to the culture environment is effectively improved.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to a microbial agent for straw fermentation padding and application of the microbial agent in healthy breeding of live pigs.
Background
The traditional pig breeding mode causes serious environmental pollution, and pig manure needs to be cleaned regularly. The pig manure not only has high nitrogen and phosphorus content, but also emits various odorous compounds, and has great harm to the environment and human health. In addition, how to ensure healthy breeding of live pigs is the second major problem in the breeding industry under the requirement that the livestock breeding in China is completely forbidden to add antibiotics. Therefore, new live pig breeding modes are urgently needed to solve the existing problems.
A technology for raising pigs by using straw fermentation padding is an organic combination of comprehensive utilization of straw and utilization of livestock and poultry manure resources, and is a breeding technology which takes crop straws of rape, corn and the like as main padding, adds crude salt, glucose powder, milk powder and special strain microorganisms, constructs a healthy micro-ecological breeding environment through fermentation and then serves as daily living padding of pigs. The technology plays an important role in solving the health problem of live pigs, can decompose excrement of the live pigs and synthesize nutrient components required by microorganisms, realizes zero discharge of the excrement, and can reduce NH in a farm3、NH3And VOCs and other odorous gases are discharged, so that quasi-zero emission of odor is realized. Can effectively reduce the disease of live pigsThe rate and the quality of pork are improved, and a culture process guarantee can be provided for the breeding innovation and the ecological culture based on local pig breeds. At present, the domestic fermentation bed microbial inoculum has the problems of high price, poor effect of treating volatile organic pollutants, poor compatibility of a deodorizing microbial inoculum and a carrier and the like. Therefore, the problem can be effectively solved by developing the microbial agent with lower cost and high efficiency, the overall technical level can reach the domestic advanced level through technical, standardization and product innovation, and the microbial agent has wide application prospect.
Disclosure of Invention
The invention aims to solve the problems in the prior art and aims to provide a microbial agent for straw fermentation padding and application of the microbial agent in healthy breeding of live pigs.
The microbial agent is designed according to the characteristics of the fermentation padding and the requirements of the padding in different periods, and is a compound flora preparation taking lactic acid bacteria, bacillus, saccharomycetes and the like as basic strains.
The composite flora preparation comprises the following components in parts by volume: 20-30 parts of lactobacillus plantarum fermentation broth, 20-30 parts of lactobacillus reuteri fermentation broth, 15-20 parts of bacillus subtilis fermentation broth, 15-20 parts of bacillus amyloliquefaciens fermentation broth and 10-20 parts of saccharomyces cerevisiae fermentation broth.
The compound flora preparation is constructed and obtained by the following method:
respectively fermenting lactobacillus plantarum and lactobacillus reuteri by a lactobacillus liquid fermentation system; respectively fermenting bacillus subtilis and bacillus amyloliquefaciens by using a bacillus liquid fermentation system; fermenting the wine brewing yeast by a yeast liquid fermentation system; after obtaining the fermentation liquor of each strain, mixing the fermentation liquor of each strain according to a proportion to obtain the composite flora preparation.
According to the invention, conditions such as initial culture medium, fermentation temperature, fermentation pH, fermentation time, rotating speed and the like are selected and optimized for three main types of lactic acid bacteria, bacillus and saccharomycetes in the flora preparation through orthogonal design, an optimal high-density preparation system is finally established, the strains are independently fermented and then mixed according to respective proportions to prepare the composite microbial inoculum, the viable count of the microbial inoculum reaches the requirement of commercialization on the density of the microbial inoculum, and the construction of the special flora preparation for raising pigs by using straw fermentation padding is completed.
The fermentation process of the optimized lactobacillus liquid fermentation system comprises the following steps: the temperature is 33-37 deg.C, pH4.0-5.5, rotation speed is 150-10。
The fermentation process of the optimized yeast liquid fermentation system comprises the following steps: temperature 25-30 deg.C, pH5.0-5.5, dissolved oxygen 25-40%, ventilation amount 2-8L/min, rotation speed 500-10。
The fermentation process of the optimized bacillus liquid fermentation system comprises the following steps: temperature is 30-37 deg.C, pH5.0-6.0, rotation speed is 120-10。
The microbial agent is applied as a fermentation bed padding in the live pig breeding process. The method is characterized in that the microbial inoculum, an auxiliary material culture medium and straws are mixed and fermented for pig breeding, and comprises the following steps:
step 1: mixing and dissolving a microbial agent and an auxiliary material culture medium in water to obtain a mixed material;
step 2: crushing the straws to 3-5cm in length, paving the crushed straws on a cement ground, uniformly spraying the mixed material obtained in the step (1) onto the surfaces of the straws, stacking the straws after uniformly stirring and mixing, covering a plastic film, compacting the periphery, and keeping a closed environment for continuous fermentation for 3-6 days.
The inoculation amount of the microbial agent is 0.1-0.5 part by weight based on 100 parts by weight of the used straw, the addition amount of the water in the step 1 is 50-150 parts by weight, and the straw is rape straw or corn straw.
Based on 100 parts by mass of the straws, the auxiliary material culture medium comprises the following components in parts by mass: 0.1-0.5 part of glucose, 0.1-0.3 part of crude salt and 0.1-0.6 part of milk powder.
According to the invention, conditions such as initial culture medium, fermentation temperature, fermentation pH, fermentation time, rotating speed and the like are selected and optimized through orthogonal design on three main types of lactic acid bacteria, bacillus and saccharomycetes in the flora preparation, an optimal high-density preparation system is finally established, the strains are independently fermented and then mixed according to respective proportions to prepare the composite microbial inoculum, the number of viable bacteria of the microbial inoculum meets the requirement of commercialization on the density of the microbial inoculum, the construction of the special flora preparation for raising pigs by using straw fermentation padding is completed, the preparation can be used for healthy culture of live pigs, the effects of zero discharge of feces and quasi zero discharge of odor are achieved, and the pollution to the culture environment is effectively improved.
Drawings
FIG. 1 shows the optimization of fermentation production conditions of a part of strains.
FIG. 2 is a graph comparing the fermentation of the straw with the bedding. Wherein the left figure is the flora preparation group of the invention, and the right figure is the aseptic agent control group.
FIG. 3 is a graph comparing the fermentation of the straw with the bedding. Wherein the left figure is the flora preparation group of the invention, and the right figure is the commercial microbial inoculum control group.
FIG. 4 shows the monitoring results of malodorous gases in the breeding house.
Detailed Description
The technical scheme of the invention is further analyzed and explained by combining specific embodiments, the invention takes rape or corn straws as raw materials to prepare the biological padding by fermentation, the preparation effect of the padding takes the growth condition of microbial spots on the surfaces of the straws as an evaluation standard, the beneficial microbial spots are uniformly distributed on the surfaces of the straws in a small and dense form, the mixed bacteria are non-uniformly distributed on the surfaces of the straws in a green or white aggregation form, and the bacterial spots are large. The following embodiments of the present invention are provided to supplement the objects, technical solutions, effects, and the like of the present invention, but the present invention is not limited to the scope of the present invention, and simple changes, modifications, or substitutions may be made within the scope of the present invention.
1. Fermentation preparation of flora preparation
On the basis of initial culture medium of lactobacillus, bacillus and yeast, the fermentation temperature (28 ℃, 30 ℃, 37 ℃ and 40 ℃), the fermentation pH (4.5, 5, 5.5, 6.4, 6.7, 7, 7.3 and 7.6), the fermentation time (12h, 24h, 26h and 48h), the stirring speed (200rpm, 400rpm, 600rpm, 800rpm and 1 rpm) and the fermentation time (12h, 24h, 26h and 48h) of three types of strains are respectively carried out by single-factor and orthogonal design000rpm), fermentation liquor dissolved oxygen (0%, 10%, 20%, 30%, 40%) and other key factors are optimized, and the optimal fermentation preparation conditions of respective strains are obtained by taking the biomass of thalli as an evaluation standard (figure 1). In this case, the CFU of the lactic acid bacteria strain fermentation broth was 6.0X 1010The CFU of the yeast strain is 3.0 multiplied by 1010The CFU of the bacillus strain is 6.0 multiplied by 1010。
2. Fermentation effect of straw padding
Respectively fermenting Lactobacillus plantarum and Lactobacillus reuteri at 37 deg.C, pH4.0 and rotation speed of 200rpm for 36 hr to obtain fermented liquid with viable count CFU of 6.0 × 1010。
Fermenting Saccharomyces cerevisiae at 30 deg.C, pH5.5, dissolved oxygen 25%, ventilation 5L/min, and rotation speed 800rpm for 36 hr to obtain yeast fermentation liquid with viable count CFU of 3.0 × 1010。
Respectively fermenting Bacillus subtilis and Bacillus amyloliquefaciens at 37 deg.C, pH6.0 and rotation speed of 200rpm for 36 hr to obtain Bacillus subtilis and Bacillus amyloliquefaciens fermentation liquid with viable count CFU of 6.0 × 1010。
And fully mixing 30 parts of lactobacillus plantarum fermentation liquid, 20 parts of lactobacillus reuteri fermentation liquid, 15 parts of bacillus subtilis fermentation liquid, 20 parts of bacillus amyloliquefaciens fermentation liquid and 15 parts of saccharomyces cerevisiae fermentation liquid to obtain the microbial composite flora preparation.
100 parts of rape straw crushed to 3-5cm in length, 0.1 part of crude salt, 0.2 part of glucose powder, 0.2 part of milk powder, 0.1 part of microbial compound flora preparation and 100 parts of water are weighed.
Dissolving crude salt, glucose powder, milk powder and compound microorganism bacterium agent in water, stirring, and mixing to obtain mixed solution.
And fully stirring and uniformly mixing the straws and the mixed solution, stacking, covering the surfaces of the straws with a plastic film, compacting the periphery, continuously keeping a closed environment, and fermenting for 3-4 days.
0.1 part of water is used for replacing the microbial inoculum, and the rest components and operation are the same, so that the straw fermentation padding of the sterile agent control group is prepared.
Comparing the flora preparation and the aseptic agent control group, the growth condition of the bacterial plaque on the surface of the straw shows that the control group has larger bacterial plaque in a green or white aggregation form and more mildew points, while the bacterial plaque of the compound microbial agent experimental group is in a small and dense form and is uniformly distributed, the fermentation effect is good, and the comparison of the change trends of the straw temperature and the pH shows that the temperature rise range and the highest value of the microbial agent group are both larger than those of the control group, the pH is rapidly reduced and is lower than those of the control group, and the temperature is rapidly increased and the pH of the fermented straw is reduced due to the heat generated by the propagation and metabolism of the thalli, so that the compound flora preparation can rapidly enter a growth and propagation metabolism stage in the preparation of the straw fermentation padding, and is beneficial to the fermentation preparation of the padding (figure 2 and table 1).
TABLE 1 fermentation parameter changes of rape stalks of compound microorganism bacterium agent test group and control group
3. Use of compound microbial agent in padding preparation
Respectively fermenting Lactobacillus plantarum and Lactobacillus reuteri at 35 deg.C, pH5.0 and rotation speed of 200rpm for 36 hr to obtain Lactobacillus plantarum and Lactobacillus reuteri fermentation liquid with viable count CFU of 6.0 × 1010。
Fermenting Saccharomyces cerevisiae at 28 deg.C, pH5.0, dissolved oxygen 30%, ventilation amount 6L/min, and rotation speed 800rpm for 36 hr to obtain yeast fermentation liquid with viable count CFU of 3.0 × 1010。
Respectively performing high-density fermentation on bacillus subtilis and bacillus amyloliquefaciens at the temperature of 35 ℃, the pH value of 6.0 and the rotation speed of 180rpm, finishing the fermentation after the fermentation is performed for 36 hours, and obtaining the bacillus subtilis and bacillus amyloliquefaciens fermentation liquid, wherein the viable count CFU of the fermentation liquid is 6.01010。
In an embodiment A, 20 parts of lactobacillus plantarum fermentation broth, 20 parts of lactobacillus reuteri fermentation broth, 20 parts of bacillus subtilis fermentation broth, 20 parts of bacillus amyloliquefaciens fermentation broth and 20 parts of saccharomyces cerevisiae fermentation broth are fully mixed to obtain the microbial composite flora preparation.
100 parts of rape straw, 0.15 part of crude salt, 0.3 part of glucose powder, 0.3 part of milk powder, 0.2 part of microbial composite flora preparation and 50 parts of water, and the rape straw fermentation padding is prepared according to the operation of the invention.
In an embodiment B, 25 parts of lactobacillus plantarum fermentation broth, 25 parts of lactobacillus reuteri fermentation broth, 15 parts of bacillus subtilis fermentation broth, 15 parts of bacillus amyloliquefaciens fermentation broth and 20 parts of saccharomyces cerevisiae fermentation broth are fully mixed to obtain the microbial composite flora preparation.
100 parts of rape straw, 0.15 part of crude salt, 0.3 part of glucose powder, 0.3 part of milk powder, 0.2 part of microbial composite flora preparation and 50 parts of water, and the rape straw fermentation padding is prepared according to the operation of the invention.
In an embodiment C, 22 parts of lactobacillus plantarum fermentation broth, 28 parts of lactobacillus reuteri fermentation broth, 20 parts of bacillus subtilis fermentation broth, 15 parts of bacillus amyloliquefaciens fermentation broth and 15 parts of saccharomyces cerevisiae fermentation broth are fully mixed to obtain the microbial composite flora preparation.
100 parts of rape straw, 0.15 part of crude salt, 0.3 part of glucose powder, 0.3 part of milk powder, 0.2 part of microbial composite flora preparation and 50 parts of water, and the rape straw fermentation padding is prepared according to the operation of the invention.
In an embodiment D, 50 parts of lactobacillus plantarum fermentation broth and 50 parts of lactobacillus reuteri fermentation broth are sufficiently mixed to obtain a microbial composite flora preparation.
100 parts of rape straw, 0.15 part of crude salt, 0.3 part of glucose powder, 0.3 part of milk powder, 0.2 part of microbial composite flora preparation and 50 parts of water, and the rape straw fermentation padding is prepared according to the operation of the invention.
According to the growth results of the microbial spots on the surfaces of the rape straws and the variation of fermentation parameters, in the embodiment A, B, C, the composite microbial spots on the surfaces of the rape straws are good in growth condition, small and dense, and uniformly distributed on the surfaces of the straws without mixed bacteria or mildew, and in the embodiment D, the composite microbial spots are relatively few in growth and partially mildew. The reproductive metabolism of the compound microbial strains generates heat to rapidly increase the temperature and reduce the pH of the fermented straws, in example A, B, C, the temperature and pH change trends are basically the same, and the temperature increase amplitude and the maximum value are higher than those of example D. The pH detection results of the straws in all the embodiments show that the embodiment D is higher than the other three groups, namely the bacteria in the embodiment grow, reproduce and metabolize slowly, and the straw padding fermentation can not be completed due to the lack of the microbial inoculum combination with the effective proportion. The results of the examples show that in the above examples, the composite microbial agent composition range specified by the present invention can complete the padding fermentation preparation of rape straws, which is beneficial to the popularization and application of the composite microbial agent (table 2).
TABLE 2 fermentation parameters of rape stalks in different use examples of the composite microbial inoculum
4. The invention relates to a contrast of a compound microbial agent and a commercial microbial agent
Respectively fermenting Lactobacillus plantarum and Lactobacillus reuteri at 37 deg.C, pH4.0 and rotation speed of 200rpm for 36 hr to obtain fermented liquid with viable count CFU of 6.0 × 1010。
Fermenting Saccharomyces cerevisiae at 30 deg.C, pH5.5, dissolved oxygen 25%, ventilation 5L/min, and rotation speed 800rpm for 36 hr to obtain yeast fermentation liquid with viable count CFU of 3.0 × 1010。
Respectively fermenting Bacillus subtilis and Bacillus amyloliquefaciens at 37 deg.C, pH6.0, and rotation speed of 200rpm for 36 hr to obtain Bacillus subtilis and Bacillus amyloliquefaciensThe viable count CFU of the bacillus fermentation liquor is 6.0 multiplied by 1010。
And fully mixing 30 parts of lactobacillus plantarum fermentation liquid, 20 parts of lactobacillus reuteri fermentation liquid, 15 parts of bacillus subtilis fermentation liquid, 20 parts of bacillus amyloliquefaciens fermentation liquid and 15 parts of saccharomyces cerevisiae fermentation liquid to obtain the microbial composite flora preparation.
100 parts of rape straw crushed to 3-5cm in length, 0.2 part of crude salt, 0.5 part of glucose powder, 0.5 part of milk powder, 0.2 part of microbial compound flora preparation and 100 parts of water are weighed.
Dissolving crude salt, glucose powder, milk powder and compound microorganism bacterium agent in water, stirring, and mixing to obtain mixed solution.
And fully stirring and uniformly mixing the straws and the mixed solution, stacking, covering the surfaces of the straws with a plastic film, compacting the periphery, continuously keeping a closed environment, and fermenting for 3-4 days.
0.2 part of commercial microbial inoculum is used for replacing the microbial inoculum of the invention, and the rest components and operation are the same, so as to prepare the straw fermentation padding of the commercial microbial inoculum test group.
Comparing the microbial composite flora preparation of the invention with commercial microbial agents sold in the market, the growth situation of microbial plaques on the surfaces of rape straws is shown, obvious plaques appear on the surfaces of two groups of test straws, the distribution is more uniform, and basically no mixed bacteria grows, which shows that the flora preparation of the invention has the fermentation effect which is not weaker than that of the commercial microbial agents sold in the market (figure 3).
5. Culture test demonstration of composite microbial inoculant fermentation bed
Laying the prepared straw padding on a fermentation bed, then carrying out a culture test demonstration, verifying and evaluating the practical application capability of a flora preparation, and carrying out NH (hydrogen) test on two pigsties by using an odor detector3、H2S, VOCs, etc. are continuously monitored. During gas detection, an air suction port of the detector is placed at a position about 3-5cm above a padding fermentation bed, and the time lasts for 1-3 minutes to stabilize the reading of the detector.
The results show that NH3、H2S, VOCs the value was low and no H could be detected in the pig house2S, NH according to the prolongation of the cultivation time3Increased concentration on day 5VOCs were detected, but their values were far below national emission standards. After the fresh padding is supplemented, all parameters are reduced, which shows that the fermentation bed microbial inoculum has better treatment effect on odor and other gases, and the optimal feeding time is set on day 5 (figure 4).
Claims (8)
1. A microbial agent for straw fermentation padding is characterized in that:
compound flora preparation using lactobacillus, bacillus, yeast, etc. as basic strains;
the composite flora preparation comprises the following components in parts by volume: 20-30 parts of lactobacillus plantarum fermentation broth, 20-30 parts of lactobacillus reuteri fermentation broth, 15-20 parts of bacillus subtilis fermentation broth, 15-20 parts of bacillus amyloliquefaciens fermentation broth and 10-20 parts of saccharomyces cerevisiae fermentation broth.
2. The microbial agent according to claim 1, which is constructed by a method comprising:
respectively fermenting lactobacillus plantarum and lactobacillus reuteri by a lactobacillus liquid fermentation system; respectively fermenting bacillus subtilis and bacillus amyloliquefaciens by using a bacillus liquid fermentation system; fermenting the saccharomyces cerevisiae by using a yeast liquid fermentation system; after obtaining the fermentation liquor of each strain, mixing the fermentation liquor of each strain according to a proportion to obtain the composite flora preparation.
3. The microbial inoculant according to claim 2, wherein:
the fermentation process of the lactobacillus liquid fermentation system comprises the following steps: the temperature is 33-37 ℃, the pH is 4.0-5.5, the rotation speed is 150-10。
4. The microbial inoculant according to claim 2, wherein:
the fermentation process of the yeast liquid fermentation system comprises the following steps: temperature is 25-30 deg.C, pH is 5.0-5.5, dissolved oxygen is 25-40%, ventilation volume is 2-8L/min, rotation speed is 500-010。
5. The microbial inoculant according to claim 2, wherein:
the fermentation process of the bacillus liquid fermentation system comprises the following steps: the temperature is 30-37 ℃, the pH is 5.0-6.0, the rotation speed is 120-10。
6. Use of a microbial inoculant according to any one of claims 1 to 5 wherein: the microbial agent is used as a fermentation bed padding in the live pig breeding process.
7. Use according to claim 6, characterized in that:
the microbial agent, an auxiliary material culture medium and straws are mixed and fermented and then are used for live pig breeding, and the method comprises the following steps:
step 1: mixing and dissolving a microbial agent and an auxiliary material culture medium in water to obtain a mixed material;
step 2: crushing the straws to 3-5cm in length, paving the crushed straws on a cement ground, uniformly spraying the mixed material obtained in the step (1) onto the surfaces of the straws, stacking the straws after uniformly stirring and mixing, covering the straws with a plastic film, compacting the periphery of the straws, and keeping a closed environment for continuous fermentation for 3-6 days.
8. Use according to claim 7, characterized in that:
the inoculation amount of the microbial agent is 0.1-0.5 part by weight based on 100 parts by weight of the used straw, the addition amount of the water in the step 1 is 50-150 parts by weight, and the straw is rape straw or corn straw.
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