CN110129233B - Microbial agent for preparing fermentation bed padding for cage-raised chickens and preparation method thereof - Google Patents
Microbial agent for preparing fermentation bed padding for cage-raised chickens and preparation method thereof Download PDFInfo
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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
- A01K31/00—Housing birds
- A01K31/04—Dropping-boards; Devices for removing excrement
-
- 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
-
- 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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Abstract
The invention discloses a microbial agent for preparing a fermentation bed padding for cage-raised chickens and a preparation method thereof. The compound microbial agent is prepared by inoculating a composition consisting of bacillus amyloliquefaciens fermentation liquid, bacillus megaterium fermentation liquid, trichoderma longibrachiatum fermentation liquid and lactobacillus plantarum fermentation liquid into a solid culture medium, uniformly mixing, culturing for 72-96h under the culture condition of 30-35 ℃, and drying. The invention discloses application of a compound microbial agent in preparation of a fermentation bed padding for caged chickens. According to the invention, four excellent strains are selected in an optimal proportion to prepare the compound microbial agent, functional strains have a synergistic effect in the feeding management process of the fermentation bed surface of the caged chicken, the functions of organic matter conversion bacteria, ammonia-fixing deodorization bacteria and bacteria inhibiting bacteria are complementary, the temperature rise fermentation of the fermentation bed is rapidly started, the rapid removal of water in excrement is realized, the microbial diversity of the fermentation bed is improved, the growth of pathogenic putrefying bacteria is inhibited, the in-situ degradation of the chicken excrement is promoted, and the humus rate of padding is improved.
Description
Technical Field
The invention discloses a microbial agent for preparing a fermentation bed padding for cage-raised chickens and a preparation method thereof, belonging to the technical field of microbial engineering.
Background
At present, a large-scale chicken farm adopts a cage-raising mode, the cage-raising mode improves the production level and the labor efficiency of the chicken farm, meanwhile, the problems of concentrated chicken manure discharge, deterioration of the environment of the chicken farm and the like exist, and manure pollution becomes an important factor restricting the cage-raising chicken breeding. Most of manure treatment technologies for cage-raised chickens adopt modes such as compost fermentation, biogas fermentation, aerobic fermentation drying technologies and the like for treatment, manure treated by compost fermentation occupies land, air pollution to the environment is large, biogas fermentation has seasonal limitation, the cost of the aerobic fermentation drying technology is high, and certain limitation is realized in actual utilization.
The fermentation bed cultivation technology is a novel ecological cultivation mode, and the key point is that fermentation padding is laid on the ground of a livestock and poultry colony house, so that the in-situ degradation of the feces and urine of the livestock and poultry is realized, the cultivation environment is greatly improved, and the method is an environment-friendly agricultural technology. The microbial fermentation bed mode realizes the synchronization of the stages of cultivation and in-situ decomposition and fermentation of excrement, and is an important way for quickly producing organic fertilizer. Humus is an important component of the organic fertilizer, and the humic acid fertilizer can obviously improve the soil fertility and promote the growth of plants. At present, the fermentation bed is mainly applied to the cultivation of live pigs, the research on the cultivation of poultry is less, and the research on the fermentation bed for cage-rearing of chickens is rarely reported.
The traditional chicken raising padding is thin and generally about 10cm, the change of chicken manure in the padding is mainly in a decay process, the padding does not have a fermentation function, the decomposition effect on the chicken manure is not large, the padding mainly has the effect of adsorbing the moisture and odor of the chicken manure, heat is not obviously generated, the padding is relatively wet, and the odor in the padding and a chicken house is serious. The thickness of the padding of the fermentation bed of the cage-raised chicken is different from that of the traditional flat-raising padding mode, and the thickness and the proportion of the padding obviously influence the porosity and the water retention of the fermentation padding. The process of fermenting and decomposing the manure by the padding is the result of the action of microorganisms, the activity of the fermentation strain determines the efficiency of decomposing the manure and generating humus, and the process is the core link of the microbial fermentation bed technology. The compound microbial agent consisting of the strains with strong colonization ability, high degradation rate and complementary functions is favorable for the in-situ degradation of the feces by the fermentation bed to form the humus organic fertilizer.
Disclosure of Invention
The invention aims to provide a microbial agent for a fermentation bed of cage-raised chickens, which can effectively promote the in-situ degradation of chicken manure, obviously reduce ammonia nitrogen and water content of padding and improve the humus rate of the padding, and a preparation method thereof.
The purpose of the invention can be realized by the following technical scheme:
a microbial composition for preparing a fermentation bed padding for cage-raised chickens is composed of microbial fermentation liquor in the following volume ratio: 20-30% of bacillus amyloliquefaciens fermentation liquid, 25-35% of bacillus megaterium fermentation liquid, 20-30% of trichoderma longibrachiatum fermentation liquid and 15-25% of lactobacillus plantarum fermentation liquid, wherein the total viable count contained in the microbial inoculum is 4 × 109-8 × 109 cfu/g.
The microbial composition is preferably prepared from the following components in percentage by volumeThe microbial fermentation liquid comprises the following components: 25% of bacillus amyloliquefaciens, 30% of bacillus megaterium, 25% of trichoderma longibrachiatum and 20% of lactobacillus plantarum, wherein the total viable count of the microbial inoculum is 4 multiplied by 10 9 -8×10 9 Between cfu/g.
1. Selection of superior strains
Fresh chicken manure contains 50% of water, 25.5% of organic matters, 1.63% of nitrogen, 1.54% of phosphorus, 0.85% of potassium, 11% of carbohydrates and 7% of fibers, and pig manure contains 60-65% of water, 15% of organic matters, 0.5% of nitrogen, 0.5-0.6% of phosphorus and 0.35-0.45% of potassium. The organic matter and nitrogen content of the chicken manure are relatively high, so that strains which can produce protease and amylase and convert ammonia nitrogen are selected to be matched with cellulase producing strains and acid producing and bacteria inhibiting strains, ammonia nitrogen in manure conversion is reduced, manure organic matter is converted, and bedding material humus is improved.
Bacillus amyloliquefaciens (CICC 20164) produces protease and amylase, can rapidly utilize organic carbon such as protein, fat, saccharide, and starch, and can promote conversion of easily degradable substances in the matrix in a short time and accelerate temperature rise of the matrix padding.
Trichoderma longibrachiatum (CICC 41185) has the advantages of rapid propagation, rapid growth and metabolism in a short time, strong capability of decomposing cellulose, and capability of producing cellulose mold koji by using bagasse, straw and mushroom bran as substrates.
The bacillus megaterium is nitrogen fixing bacillus with nitrification function screened from chicken manure, and the strain is preserved in China center for type culture Collection with the preservation address: in Wuhan university school of Wuhan 299 eight-way in Wuhan district, wuhan city, hubei province, the eight schools are named in classification: bacillus megaterium, with a deposit number: CCTCC No of M2019050, and the preservation date is as follows: 2019, month 1 and 30. The surface of the bacterial colony is smooth and opaque, the bacterial colony is dirty white, the cell is a long rod-shaped gram-positive bacterium, spores are formed after the bacterial colony is cultured for 2 to 3 days at 37 ℃, the size of each spore is 0.6 to 1.5 mu m, and the spores are oval.
The lactobacillus plantarum is lactobacillus screened from straws, has high acid production speed and strong growth capacity, and can inhibit pathogenic bacteria; the strain is preserved in China center for type culture Collection with the preservation addresses: in the Wuhan university school of Wuhan 299 eight roads in Wuhan city, hubei province, the classification and naming are as follows: lactobacillus plantarum, accession number: CCTCC No. M2016281, preservation date is: 2016, 5 months and 25 days. The bacterial colony of the lactobacillus plantarum is milky white, and the edge is neat; the microscopic morphology of the cells is short rod-shaped, has no spores and is gram-positive; catalase negative, oxidase negative; has strong growth and acid production performance, can grow on a culture medium with the initial pH value of 3.0-8.0, shows weak growth at 10 ℃, has good growth condition at 15-45 ℃, and has good growth condition under the condition of not more than 6.5 percent of NaCl.
Preparation of the microbial composition of the invention:
1. first-stage expanding culture: taking out purified and cultured strains (bacillus amyloliquefaciens, bacillus megaterium, trichoderma longibrachiatum and lactobacillus plantarum) from the preserved inclined plane, respectively inoculating the strains into five prepared seed culture media for respective culture, respectively filling 200ml of the culture media into each 1L bottle, culturing the lactobacillus plantarum in a constant-temperature incubator at 37 ℃ for 48h, performing shaking culture on the bacillus amyloliquefaciens and the bacillus megaterium on a shaking table at 35 ℃ and 150r/min for 48h, and performing shaking culture on the trichoderma longibrachiatum on the shaking table at 30 ℃ and 120r/min for 96h.
2. Secondary expanding culture: during the period, the number of live bacteria in the culture liquid of each bacterium is respectively detected until the number of live bacteria in each bacterium is more than or equal to 2 multiplied by 10 8 After the concentration of the microorganism strain is completed, taking out seed liquid, and preparing the microorganism composition by the four bacteria according to the strain proportion.
The compound microbial agent is prepared by inoculating the microbial composition into a solid culture medium, uniformly mixing, culturing for 72-96h under the culture condition of 30-35 ℃, and drying.
Preferably, the composite microbial agent is prepared by inoculating the microbial composition into a solid culture medium according to the weight ratio of 1; then drying at 50 ℃ to prepare the compound microbial agent; the formula of the solid culture medium is as follows: 64% of soybean curd residue, 22% of bran and corn flour10 percent of glucose, 3.5 percent of NaCl, 0.4 percent of KH 2 PO 4 0.1％、pH6.4-7.2。
The preparation method of the compound microbial agent comprises the steps of inoculating the microbial composition into a solid culture medium, uniformly mixing, culturing for 72-96h under the culture condition of 30-35 ℃, and drying to obtain the compound microbial agent; preferably, the microbial composition is inoculated in a solid culture medium, uniformly mixed and cultured for 72-96h under the culture condition of 30-35 ℃; then drying at 50 ℃ to prepare the compound microbial agent.
The formula of the solid culture medium is preferably as follows: 64% of bean curd residue, 22% of bran, 10% of corn flour, 3.5% of glucose, 0.4% of NaCl and KH 2 PO 4 0.1 percent and pH6.4-7.2; the weight ratio of the microbial composition according to the invention to the solid medium is preferably 1.
The invention relates to application of a compound microbial agent in preparation of a fermentation bed padding for cage-raised chickens.
A fermentation bed padding for caged chickens comprises the composite microbial agent.
The padding of the fermentation bed for the cage-raised chickens preferably further comprises mushroom bran, sawdust, rice hulls and bran.
The fermentation bed padding for the cage-raised chickens is further preferably prepared by the following method: laying 35-40 cm of fungus chaff on a fermentation bed of the caged chicken, laying 8-15 cm of mixture of wood chips and rice hulls with the same weight ratio on the surface of the fermentation bed, stirring by using a rake-turning machine, and mixing the fungus chaff with the mixture of the wood chips and the rice hulls; uniformly mixing the composite microbial agent and the bran of claim 3 or 4, scattering the mixture on the surface of a fermentation bed, and then stirring the mixture by using a rake-turning machine; the weight ratio of the composite microbial agent to the bran is as follows: 1; 50g of the compound microbial agent is scattered on each square of padding.
The invention has the beneficial effects that:
(1) The compound microbial agent is prepared by selecting four excellent strains according to an optimal proportion, functional strains can perform a synergistic effect in a feeding management process of a fermentation bed surface of a caged chicken, organic matter conversion bacteria (bacillus amyloliquefaciens and trichoderma longibrachiatum), ammonia fixing deodorization bacteria (bacillus megaterium) and germ inhibiting bacteria (lactobacillus plantarum) are complementary in function, the temperature rise fermentation of the fermentation bed is quickly started, the moisture in excrement is quickly removed, the microbial diversity of the fermentation bed is improved, the growth of pathogenic putrefying bacteria such as staphylococcus aureus and epidemic bacillus is inhibited, and the in-situ degradation of the chicken excrement is effectively promoted.
(2) The bacillus megaterium separated from the chicken manure can be fixedly planted and rapidly grow in a cage chicken fermentation bed, so that ammonia nitrogen of padding materials is remarkably reduced, ammonia gas is reduced, and nitrogen loss is reduced in the temperature rise process.
(3) The bedding combination of the fungus chaff, the rice hull and the wood dust is matched with the compound microbial agent, so that the content of humus such as humic acid can be obviously improved, the humus rate of the bedding can be improved, and a high-quality organic fertilizer is formed.
Drawings
FIG. 1 differential analysis of the abundance of bacterial colonies in different litter sizes
Biological material preservation information
The Bacillus megatherium BM18 is classified and named as Bacillus megaterium BM18 and is preserved in China center for type culture Collection with the preservation number: CCTCC No. M2019050, and the preservation date of the university of Wuhan, china is 1 month and 15 days in 2019. Lactobacillus plantarum ZRR, classified and named Lactobacillus plantarum ZRR, deposited in China center for type culture Collection with the deposit number: CCTCC No. M2016281, preservation address of Wuhan university in Wuhan, china, and preservation date of 2016, 5 and 25 days.
Detailed Description
The following examples are intended to further illustrate the invention, but not to limit it.
The name and source of the 4 strains adopted by the invention are as follows:
bacillus amyloliquefaciens (CICC 20164) and Trichoderma longibrachiatum (CICC 41185) are purchased from China center for Industrial culture Collection of microorganisms. The bacillus megaterium, the nitrogen-fixing bacillus with nitrification function screened from chicken manure, has been preserved in China center for type culture Collection with the preservation number: CCTCC No. M2019050. Lactobacillus plantarum is lactic acid bacteria which is screened from straws, has high acid production speed, strong growth capacity and pathogenic bacteria inhibition, is preserved in the China center for type culture Collection with the preservation number: CCTCC No. M2016281.
Example 1: isolation of Bacillus megaterium and Lactobacillus plantarum
Isolation of Bacillus megaterium: 10g of decomposed chicken manure is inoculated into 200mL of enrichment medium (2 g of ammonium sulfate, 2.17g of sodium succinate, 0.25g of monopotassium phosphate, 0.125g of magnesium sulfate, 0.125g of NaC and 1000mL of water), the mixture is transferred into a new enrichment medium at the inoculation amount of 3% every 3d, the mixture is continuously enriched and cultured for 30d, diluted and coated on an ammonium sulfate plate, large colonies with fast growth are selected, and the separation and purification are repeated twice. Inoculating the obtained strain in enrichment medium, shaking for culture, tracking and detecting nitrite and nitrate production conditions with Grignard reagent and diphenylamine reagent to obtain 28 strains, and inoculating the strains into chicken manure leachate (preparation of chicken manure leachate: mixing chicken manure and tap water at a ratio of l: 1), sterilizing at 121 deg.C for 20min in autoclave, standing overnight, filtering, and sterilizing at 12l deg.C for 20 min). The content of ammonia nitrogen before and after inoculation is measured by adopting a potassium chloride leaching-indophenol blue colorimetric method to obtain 6 bacterial strains with strong degradation capability, wherein the highest ammonia nitrogen degradation rate of one bacterial strain in 48 hours reaches 69.7%. The bacterial colony of the strain is large, and the edge is irregular; the thallus is rod-shaped, has spores and is gram-positive; the bacillus megaterium is identified as bacillus megaterium by a 16rRNA gene sequence and is preserved in China center for type culture collection with the preservation number: CCTCC No. M2019050.
And (3) separating the lactobacillus plantarum: weighing 10g of straw sample, adding the straw sample into 90ml of sterilized distilled water, coating the straw sample on an MRS solid culture medium after gradient dilution, culturing for 48h, selecting a bacterial colony growing on the MRS according to form and color, screening 1 strain of lactic acid bacteria which can produce acid, have strong acid resistance, acid resistance and stress resistance and can inhibit pathogenic bacteria through tests such as acid production, acid resistance, temperature test and the like, wherein the lactic acid yield is 1.51mg/L in 24 hours, and the inhibition zones for Escherichia coli K88 and Staphylococcus aureus are 21.4mm and 20.8mm respectively. The bacterial colony is milky white, and the edge is neat; the microscopic morphology of the cells is short rod-shaped, no spores and gram-positive; catalase negative, oxidase negative; the lactobacillus plantarum has stronger growth and acid production performance, can grow on a culture medium with the initial pH value of 3.0-8.0, shows weak growth at 10 ℃, has good growth condition at 15-45 ℃, is identified as lactobacillus plantarum through a 16rRNA gene sequence, is preserved in the China center for type culture Collection, and has the preservation number: CCTCC No. M2016281.
Example 2 preparation of Complex microbial Agents
The amplification of the compound microbial inoculum from slant strains to finished products is carried out by the following three steps
1. First-stage expanding culture: taking out purified and cultured strains (bacillus amyloliquefaciens, bacillus megaterium, trichoderma longibrachiatum and lactobacillus plantarum) from the preserved inclined plane, respectively inoculating the strains into four prepared seed culture media for respective culture, respectively filling 200ml of the culture media into each bottle with 1L, culturing the lactobacillus plantarum in a constant-temperature incubator at 37 ℃ for 48h, performing shake culture of the bacillus amyloliquefaciens and the bacillus megaterium on a shaking table at 35 ℃ and 150r/min for 48h, and performing shake culture of the trichoderma longibrachiatum on the shaking table at 30 ℃ and 120r/min for 96h.
In the first-stage propagation, the slant medium and the seed medium of each bacterium are known as basal media. Wherein the seed culture medium of bacillus amyloliquefaciens and bacillus megaterium-nutrient broth culture medium (culture bacteria), the trichoderma longibrachiatum-PDA culture medium (culture mould) and the lactobacillus plantarum-MRS culture medium (culture lactobacillus).
2. Secondary expanding culture: during the period, the number of live bacteria in each bacteria culture solution is respectively detected until the number of live bacteria in each bacteria culture solution is more than or equal to 2 multiplied by 10 9 After the solution is subjected to the concentration and the concentration, the seed solution is taken out, the four bacteria are prepared into compound bacteria solution according to the volume percentage of 25 percent of bacillus amyloliquefaciens, 30 percent of bacillus megaterium, 25 percent of trichoderma longibrachiatum and 20 percent of lactobacillus plantarum, then the compound bacteria solution is inoculated into a solid culture medium according to the weight ratio of 10 percent of the compound bacteria solution to 90 percent of the solid culture medium, and the compound bacteria solution is uniformly mixed and is placed under the culture condition of 35 ℃ for culture for 72 hours.
The solid culture medium formula (weight percentage) in the second-stage propagation: 64% of bean curd residue (water content 80%) and 22% of bran10% of corn flour, 3.5% of glucose, 0.4% of NaCl and KH 2 PO 4 0.1％、pH6.4-7.2。
3) Solid material drying: drying at 50 deg.C to obtain compound microbial preparation. The bacterial content of the microbial inoculum is 4 multiplied by 10 9 -8×10 9 Between cfu/g
Example 3: bed body effect test of the compound microbial leavening agent in the fermentation bed of the caged chicken
Application experiments of the product on a bed surface substrate are carried out in a chicken farm from 10 months in 2017 to 12 months in 2017. The method comprises the following steps of laying mushroom bran of about 40cm on a fermentation bed of the caged chicken, laying sawdust/rice hulls of about 10cm on the surface, wherein the weight ratio of the sawdust to the rice hulls is 1. The test group uses 50 grams of fermentation bed microbial inoculum for each square meter of fermentation bed, and the spreading method comprises the steps of firstly, uniformly mixing the fermentation bed microbial inoculum and bran, then, spreading the mixture on the surface of the fermentation bed, and stirring the mixture by using a rake-turning machine; the fermentation bed microbial inoculum and the bran are in the following weight ratio: 50g of fermentation bed microbial inoculum and 1kg of bran. The control group was dusted with 1kg of bran per square meter.
Taking fermentation bed padding at 15d, 30d, 45d and 60d respectively, and monitoring the microbial and nutrient contents of the padding. Culturing total bacteria in Nutrient Agar (NA) culture medium at 37 deg.C for 24 hr; culturing the diluted solution of bacillus in water bath at 80 deg.C for 10min, and culturing in Nutrient Agar (NA) culture medium at 37 deg.C for 24 hr; culturing the fungus in potato glucose agar culture medium at 28 deg.C for 5 days; staphylococcus aureus and Escherichia coli were cultured in mannitol sodium chloride agar (MSA) and eosin methylene blue agar medium (EMB) at 37 deg.C for 48 hr. The nitrifying bacteria are treated by Stephenson culture solution MPN method, and the denitrifying bacteria are treated by sodium citrate potassium nitrate culture medium MPN method. The ammoniacal nitrogen is measured by potassium chloride extraction-indophenol blue colorimetric method. Organic matters are measured by a potassium dichromate oxidation method. The humic substances and components of the padding are determined by adopting a sodium pyrophosphate-sodium hydroxide solution extraction method and a potassium dichromate method. The results are shown in tables 1, 2 and 3. As can be seen from tables 1 and 2, the addition of the microbial inoculum of the invention obviously improves the distribution quantity of bacillus, obviously reduces the distribution quantity of staphylococcus aureus, obviously improves nitrobacteria (P is less than 0.05) in nitrogen metabolism microorganisms, and reduces nitrogen loss. As can be seen from Table 3, the addition of the microbial inoculum of the invention can significantly reduce the water content and the ammonia nitrogen content of the padding, thereby significantly improving the environment of the poultry house, and in the aspect of making the padding into fertilizer, the addition of the microbial inoculum significantly increases the contents of humic acid and total humus, increases the humus rate of the padding, and forms a high-quality organic fertilizer.
TABLE 1 bedding microorganisms of fermentation bed for caged chicken (LogCFU/g)
TABLE 2 Nitrogen-metabolizing microorganisms (LogCFU/g) of fermentation bed litter for caged chickens
TABLE 3 nutrient content of fermentation bed padding for caged chicken
In order to further research the microbial area system of the fermentation bed of the caged chicken, the microbial composition of the caged chicken padding is systematically researched by adopting a molecular microbiology method, the microbial DNA of a 60d padding sample is extracted, the 16SrDNAV3 area is amplified, and the HiSeq platform is adopted for sequencing analysis. The results are shown in table 4 and fig. 1. The shannon diversity index and the simpson index are important indexes reflecting the microbial diversity. The higher the shannon diversity index, the lower the simpson index, indicating a higher diversity of microorganisms. As can be seen from Table 4, the addition of the fermentation bed microbial inoculum significantly improves the diversity of chicken litter microorganisms and significantly promotes the in-situ degradation of chicken manure and urine by the microorganisms. LEfSe analysis is carried out on the DNA of the padding microorganisms of different groups, and the abundance change of the dominant flora of the two groups is searched. The results are shown in fig. 1, the addition of the chicken fermentation bed microbial inoculum remarkably improves the abundance of flora of Bacillus (Bacillus) and marine Bacillus (oceanobacter) in the bedding material, and remarkably reduces the abundance of flora of epidemic Bacillus (Vulgatibacter). Bacillus and marine Bacillus are important bacterial groups involved in fecal transformation. Test results show that by adding the fermentation bed microbial inoculum, bacillus in the microbial inoculum can quickly grow in the chicken manure padding to form dominant flora, so that in-situ degradation of chicken manure is promoted.
Table 4 microbial diversity analysis of different litter samples
Claims (11)
1. A microbial composition for preparing a fermentation bed padding for cage-raised chickens is characterized by consisting of microbial fermentation liquor in the following volume ratio: 20-30% of bacillus amyloliquefaciens CICC 20164 fermentation broth, 25-35% of bacillus megaterium fermentation broth with the preservation number of CCTCC No: M2019050, 20-30% of trichoderma longibrachiatum CICC 41185 fermentation broth, 15-25% of lactobacillus plantarum fermentation broth with the preservation number of CCTCC No: M2016281, and the total viable count of the microbial inoculum is 4 x 10 9 -8×10 9 Between cfu/g.
2. The microbial composition of claim 1, consisting of the following volume ratios of microbial fermentation broth: 25% of bacillus amyloliquefaciens, 30% of bacillus megaterium, 25% of trichoderma longibrachiatum and 20% of lactobacillus plantarum, wherein the total viable count of the microbial inoculum is 4 multiplied by 10 9 -8×10 9 Between cfu/g.
3. A compound microbial agent for preparing a fermentation bed padding for cage-raised chickens is characterized in that the compound microbial agent is prepared by inoculating the microbial composition of claim 1 or 2 into a solid culture medium, uniformly mixing, culturing for 72-96h under the culture condition of 30-35 ℃, and drying.
4. The complex microbial inoculant according to claim 3, wherein the microbial composition according to claim 1 or 2 and the solid culture medium are inoculated in the solid culture medium according to the weight ratio of 1; then drying at 50 ℃ to prepare the compound microbial agent; the formula of the solid culture medium is as follows: 64% of bean curd residue, 22% of bran, 10% of corn flour, 3.5% of glucose, 0.4% of NaCl, 0.1% of KH2PO4 and pH6.4-7.2.
5. The preparation method of the compound microbial agent of claim 3, which is characterized by comprising the steps of inoculating the microbial composition of claim 1 or 2 into a solid culture medium, uniformly mixing, culturing for 72-96h under the culture condition of 30-35 ℃, and drying to obtain the compound microbial agent.
6. The method according to claim 5, wherein the solid medium is prepared by the following formula: 64% of bean curd residue, 22% of bran, 10% of corn flour, 3.5% of glucose, 0.4% of NaCl, 0.1% of KH2PO4 and pH6.4-7.2; the microbial composition of claim 1 or 2 in a weight ratio to solid medium of 1.
7. The use of the complex microbial inoculant defined in claim 3 or claim 4 in the preparation of a fermentation bed padding for a caged chicken.
8. A fermentation bed padding for caged chickens, characterized by comprising the complex microbial agent of claim 3 or 4.
9. The fermentation bed padding for caged chicken as claimed in claim 8, wherein the fermentation bed padding for caged chicken further comprises mushroom bran, wood chips, rice hulls and bran.
10. The fermentation bed padding for the cage-raised chickens, as claimed in claim 8, is characterized in that the fermentation bed for the cage-raised chickens is paved with 35-40cm of fungus chaff, the surface of the fungus chaff is paved with 8-15cm of mixture of sawdust and rice husk in equal weight ratio, a rake-turning machine is used for stirring, the fungus chaff and the mixture of the sawdust and the rice husk are mixed, the composite microbial agent and the bran as claimed in claim 3 or 4 are uniformly mixed, then the mixture is scattered on the surface of the fermentation bed, and then the rake-turning machine is used for stirring.
11. Wherein the weight ratio of the compound microbial inoculum to the bran of claim 3 or 4 is as follows: 1 to 15-22, and scattering 50g of the composite microbial agent disclosed in claim 3 or 4 per square padding.
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