CN104862298A - Composite microbial starter and preparation method thereof - Google Patents

Abstract

The invention discloses a composite microbial starter and a preparation method thereof, aiming at solving the technical problems of high efficiency and comprehensive fermentation and composting treatment of organic wastes. The composite microbial leaven consists of carrier and composite bacteria, the carrier is natural zeolite powder, and the composite bacteria consists of bacillus subtilis, lactobacillus acidophilus, lactobacillus delbrueckii subsp bulgaricus, saccharomyces cerevisiae, bacillus megaterium and clostridium pasteurianum. The preparation method comprises the following steps: inoculating, carrying out primary culture and secondary culture, mixing to obtain a compound bacterial liquid, adsorbing the compound bacterial liquid on natural zeolite powder with the granularity of 200 meshes, and drying to obtain the compound microbial starter. Compared with the prior art, the invention can decompose the organic waste efficiently and comprehensively, thoroughly solve the pollution of the organic waste to the environment, improve the resource attribute of the organic waste, and prepare the multifunctional organic fertilizer which provides comprehensive nutrients required by plant growth and well improves the soil.

Description

Composite microbial starter and preparation method thereof

technical field

The invention relates to an organic fertilizer and a preparation method thereof, in particular to a microbial fermentation agent of an organic fertilizer and a preparation method thereof.

Background technique

According to the current two-way development model of agricultural industrialization and urbanization in cities and towns, the amount of organic solid waste produced is increasing day by day. From the perspective of agricultural industrial structure, the main source of agricultural waste comes from animal husbandry and planting. On the other hand, urban household garbage contains a large amount of organic waste, such as kitchen waste and sludge produced every day by sewage treatment plants. The organic waste produced by animal husbandry and planting has the value of recycling. Among them, poultry manure contains a large number of elements such as nitrogen, phosphorus, and potassium needed for crop growth, and also contains calcium, magnesium, sulfur, zinc, boron, molybdenum, copper, Microelements such as iron. Planting straw and slag contain a lot of organic matter. The components of food waste are mainly degradable organic matter, the main components are starch contained in staple food, cellulose and polypentose contained in vegetables and plant stems and leaves, protein and fat contained in meat, and monosaccharide contained in fruit. , fruit acid and pectin etc. The sludge produced by sewage treatment plants every day contains a large number of elements such as nitrogen, phosphorus, and potassium needed for crop growth, as well as rich organic matter. The nutrients contained in the above organic wastes have the value of recycling. After inoculation, fermentation and composting, they can be made into organic fertilizers, which can be used as nutrients for plant growth, improve soil structure, increase soil fertility, and promote crop growth. However, in the existing technology, the above-mentioned different types of organic wastes are often fermented separately and then formulated into organic fertilizers, which not only has few functions, low fertilizer efficiency, and low comprehensive utilization rate of organic wastes, but also cannot effectively kill harmful pathogenic bacteria, which is very smelly , unfavorable to the health of agricultural practitioners.

Contents of the invention

The purpose of the present invention is to provide a compound microbial fermentation agent and its preparation method. The technical problem to be solved is to efficiently and comprehensively ferment and compost organic waste, improve fertilizer efficiency, and protect the health of agricultural practitioners.

The present invention adopts the following technical scheme: a kind of composite microbial fermentation agent, is made up of carrier and composite bacterial classification, is mixed according to the mass ratio of 1-3:1 of the composite bacterial liquid of carrier and composite bacterial classification, obtains after drying, and described carrier is The natural zeolite powder with a particle size of 200 meshes, the composite bacterial liquid is composed of the mass percent of the following bacterial strains: Bacillus subtilis 20-30%, Lactobacillus acidophilus 15-20%, Lactobacillus delbrueckii subspecies bulgaricus 15-20% %, 10-15% of Saccharomyces cerevisiae, 10-15% of Bacillus megaterium, 10-15% of Clostridium pasteurianus, and the number of bacteria per milliliter of the carrier is 200-300 million.

The composite bacterial liquid of the present invention is composed of the following bacterial liquid in mass percent: 25% of Bacillus subtilis, 15% of Lactobacillus acidophilus, 15% of Lactobacillus delbrueckii subspecies bulgaricus, 15% of Saccharomyces cerevisiae, 15% of Bacillus megaterium, 15% of Clostridium sporogenes, and the number of bacteria per milliliter of carrier is 200 million.

A preparation method for a composite microbial starter, comprising the following steps:

1. Inoculation: Bacillus subtilis, Lactobacillus acidophilus, Lactobacillus delbrueckii subspecies bulgaricus, Saccharomyces cerevisiae, Bacillus megaterium and Clostridium pasteurianus were cultured for 16-20 hours at room temperature with a relative humidity of 50%. Then each was added into sterile water and shaken for 30 minutes to obtain the bacterial solution;

Two, primary culture, inoculate each bacterial liquid described in the conical flask filled with primary liquid culture medium respectively, the inoculum size is 5%-10%, the pH value of culture medium is 6.0-8.0, with 180r/min Cultivate at a rotating speed for 24-48 hours to obtain a first-level expansion culture solution, and the number of bacteria in the first-level expansion culture solution reaches 200-300 million/ml;

3. Secondary culture: Mix each first-level expansion culture liquid with the second-level liquid medium and inoculate it into the Erlenmeyer flask containing the second-level liquid medium. Cultivate at min speed for 48-72 hours to obtain the secondary expansion culture liquid, and the number of bacteria in the secondary expansion culture liquid reaches 200-300 million/ml respectively;

4. According to the mass percentage content of the secondary expansion culture liquid of various strains: Bacillus subtilis 20-30%, Lactobacillus acidophilus 15-20%, Lactobacillus delbrueckii subspecies bulgaricus 15-20%, Saccharomyces cerevisiae 10-15%, Bacillus megaterium 10-15%, Clostridium pasteurianus 10-15%, mixed to obtain a composite bacterial solution; the composite bacterial solution is adsorbed on the natural zeolite powder with a particle size of 200 mesh, and the natural zeolite powder with a particle size of 200 mesh and The mass ratio of the composite bacterial solution with the composite bacterial species is 1-3:1, and drying to obtain a composite microbial starter.

In the second step of the method of the present invention, the inoculation amount is 10%, and the pH value of the culture medium is 6.0-7.0; the culture is cultivated for 32 hours at a speed of 180 r/min.

The number of bacteria in the second step of the method of the present invention is 200 million/ml.

The first-level liquid culture medium of the step 2 of the inventive method consists of respectively:

(1) The primary liquid culture medium of Bacillus subtilis consists of: 5.0g peptone, 3.0g beef extract, 5.0g NaCl, 15.0g agar, 1.0L distilled water; after mixing, sterilize at 112°C for 20min, cool naturally to room temperature, pH7.0;

(2) Lactobacillus acidophilus primary liquid medium consists of: 10.0g casein peptone, 10.0g beef extract, 5.0g yeast powder, 5.0g glucose, 5.0g sodium acetate, 2.0g diammonium citrate, and sorbitan Monooleate polyoxyethylene ether 1.0g, K ₂ HPO ₄ 2.0g, MgSO ₄ .7H ₂ O 0.2g, MnSO ₄ .H2O 0.05g, CaCO ₃ 20.0g, agar 15.0g, distilled water 1.0L; after mixing 112 Sterilize at ℃ for 20min, cool naturally to 37℃, pH6.8;

(3) The primary liquid culture medium of Lactobacillus delbrueckii subsp. 801.0g, K ₂ HPO ₄ 2.0g, MgSO ₄ .7H ₂ O 0.2g, MnSO ₄ .H ₂ O 0.05g, CaCO ₃ 20.0g, agar 15.0g, distilled water 1.0L; after mixing, sterilize at 112°C for 20min, naturally Cool to 37°C, pH6.8;

(4) Saccharomyces cerevisiae primary liquid medium consists of: 5 ° Bé wort 1.0 L, agar 15.0 g; after mixing, sterilize at 112 ° C for 20 minutes, naturally to 28 ° C, natural pH;

(5) The composition of Bacillus megaterium primary liquid medium is: 5.0g peptone, 3.0g beef extract, 5.0g NaCl, 15.0g agar, 1.0L distilled water; after mixing, sterilize at 112°C for 20min, and cool naturally to 30°C , pH7.0;

(6) The first-grade liquid culture medium of Clostridium pasteurianus consists of: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L; after mixing, sterilize at 112°C for 20min, and cool naturally to culture Temperature 37°C, pH 7.0.

In step 3 of the method of the present invention, the inoculum amount is 10%; the cultivation is carried out at 180 r/min for 48 hours.

The number of bacteria in the secondary expansion culture liquid of step 3 of the method of the present invention is respectively 200 million/ml.

The composition of the secondary liquid medium in Step 3 of the method of the present invention is: 10.0g of peptone, 10.0g of beef extract, 15.0g of agar, 5.0g of molasses, 0.2g of MgSO ₄ .7H ₂ O, 0.05g of MnSO ₄ .H ₂ O, CaCO ₃ 20.0g, NaCl 5.0g, distilled water 1.0L; after mixing, sterilize at 112°C for 20min, cool naturally to room temperature, and adjust the natural pH.

Step 4 of the method of the present invention The mass percentage content of the secondary expansion culture liquid of each strain is: 25% of Bacillus subtilis, 15% of Lactobacillus acidophilus, 15% of Lactobacillus delbrueckii subsp. bulgaricus, 15% of Saccharomyces cerevisiae, Bacillus megaterium 15%, Clostridium pasteurianus 15%.

Compared with the prior art, the present invention can efficiently and comprehensively decompose organic wastes, solve the environmental pollution caused by organic wastes, improve the resource properties of organic wastes, turn waste into treasure, and use the compound microorganisms The multifunctional organic fertilizer made of starter provides the comprehensive nutrients needed for plant growth and improves the soil well.

Detailed ways

The present invention is described in further detail below in conjunction with embodiment. Composite microbial fermentation agent of the present invention, is made up of carrier and composite bacterial classification, mixes by the mass ratio 1-3:1 of the composite bacterial liquid of carrier and composite bacterial classification, obtains after drying, each milliliter bacterial liquid of each milliliter carrier 2- 300 million bacteria count, preferably 200 million bacteria count. The carrier is natural zeolite powder with a particle size of 200 mesh. The strains are: Bacillus subtilis (Bacillus substilis, strain number CICC23013, purchased from China Industrial Microbiology Culture Collection Management Center), Lactobacillus acidophilus (Lactobacillus acidophilus, strain number CICC22150, purchased from China Industrial Microbiology Culture Collection Management Center) , Lactobacillus delbrueckii subsp.bulgaricus (Lactobacillus delbrueckii subsp.bulgaricus, strain number CICC22153, purchased from China Industrial Microorganism Culture Collection Management Center), Saccharomyces cerevisiae (Saccharomyces cerevisiae, strain number CICC32390, purchased from China Industrial Microbiology Culture Collection Management Center ), Bacillus megaterium (strain number CICC21446, purchased from China Industrial Microorganism Culture Collection Management Center) and Clostridium pasteurianum (Clostridium pasteurianum, strain number CGMCC1.2675, purchased from China General Microbiology Culture Collection Management Center ). The mass percent content of each bacterial strain in the composite bacterial liquid is: 20-30% of Bacillus subtilis, 15-20% of Lactobacillus acidophilus, 15-20% of Lactobacillus delbrueckii subsp. -15%, Bacillus megaterium 10-15%, Clostridium pasteurianus 10-15%. The preferred mass percentages of various bacterial strains are: 25% of Bacillus subtilis, 15% of Lactobacillus acidophilus, 15% of Lactobacillus delbrueckii subsp. bulgaricus, 15% of Saccharomyces cerevisiae, 15% of B. Clostridium sporogenes 15%.

The preparation method of composite microorganism starter of the present invention may further comprise the steps:

1. Inoculation, according to the temperature and humidity range of the clean room and the cleanliness level of suspended particles in the air stipulated in the standard "Code for Design of Clean Plants" GB50073-2001, respectively Bacillus subtilis, Lactobacillus acidophilus, Lactobacillus delbrueckii Subspecies, Saccharomyces cerevisiae, Bacillus megaterium and Clostridium pasteuriani were respectively inoculated on slant surfaces according to the prior art, at room temperature (20°C), relative humidity was 50%, cultivated for 16-20h, and then each was added to sterile water After shaking for 30 minutes, the bacterial liquid was obtained, counted with a hemocytometer, and the number of bacteria in the bacterial liquid reached 200-300 million/ml, preferably 200 million/ml.

Two, one-level cultivation (one-level expansion cultivation), the bacterial classification of each bacterial liquid described is inoculated in the 50ml Erlenmeyer flask that fills one-level liquid culture medium respectively, inoculum size 5%-10% (v/v), Preferably 10% (v/v), the pH value of the medium is 6.0-8.0, preferably 6.0-7.0, the Erlenmeyer flask is placed in a shaker at 180r/min, and the shaking speed is cultivated for 24-48h, preferably For 32h, obtain the first-level expansion culture liquid, count with a hemocytometer, and the number of bacteria in the first-level expansion culture liquid reaches 200-300 million/ml, preferably 200 million/ml.

The composition of primary liquid culture medium is respectively:

(1) The primary liquid culture medium for Bacillus subtilis consists of: 5.0g peptone, 3.0g beef extract, 5.0g NaCl, 15.0g agar, and 1.0L distilled water. After mixing, sterilize at 112°C for 20 minutes, and cool naturally to room temperature, pH7.0. Beef extract (Beaf extract) is a brownish-yellow to tan paste obtained from fresh beef after removing fat, digesting, filtering, and concentrating. It has a natural aroma of beef and is easily soluble in water. The aqueous solution is light yellow. Also known as beef extract, beef extract, beef extract, beef powder, beef extract powder, or beef extract powder.

(2) Lactobacillus acidophilus primary liquid medium consists of: 10.0g casein peptone, 10.0g beef extract, 5.0g yeast powder, 5.0g glucose, 5.0g sodium acetate, 2.0g diammonium citrate, and sorbitan Monooleate polyoxyethylene ether (Tween 80) 1.0g, K ₂ HPO ₄ 2.0g, MgSO ₄ .7H ₂ O 0.2g, MnSO ₄ .H2O 0.05g, CaCO ₃ 20.0g, agar 15.0g, distilled water 1.0L . After mixing, sterilize at 112°C for 20 minutes, cool naturally to 37°C, pH 6.8.

(3) The primary liquid culture medium of Lactobacillus delbrueckii subsp. 801.0g, K ₂ HPO ₄ 2.0g, MgSO ₄ .7H ₂ O 0.2g, MnSO ₄ .H ₂ O 0.05g, CaCO ₃ 20.0g, agar 15.0g, distilled water 1.0L. After mixing, sterilize at 112°C for 20 minutes, and cool naturally to 37°C. pH6.8.

(4) The primary liquid culture medium of Saccharomyces cerevisiae consists of: 1.0L of 5°Bé wort, 15.0g of agar. After mixing, it was sterilized at 112°C for 20 minutes, and naturally cooled to 28°C, with natural pH.

(5) The primary liquid culture medium for Bacillus megaterium consists of: 5.0 g of peptone, 3.0 g of beef extract, 5.0 g of NaCl, 15.0 g of agar, and 1.0 L of distilled water. After mixing, sterilize at 112°C for 20 minutes, cool naturally to 30°C, pH 7.0.

(6) The primary liquid medium of Clostridium pasteurianus consists of: 10.0 g peptone, 3.0 g beef extract, 5.0 g NaCl, 15.0 g agar, and 1.0 L distilled water. After mixing, sterilize at 112°C for 20 minutes, and cool naturally to the culture temperature of 37°C, pH 7.0.

The role of primary culture is to make expansion culture liquid for secondary culture.

3. Secondary cultivation (secondary expansion): each first-level expansion culture liquid is mixed with the second-level liquid medium and inoculated into the 1000ml Erlenmeyer flask containing the second-level liquid medium, and the inoculum size is 5%. -10% (v/v), preferably 10% (v/v), the Erlenmeyer flask is placed in a shaker and cultured at a shaking speed of 180r/min for 48-72h, preferably 48h, to obtain the secondary expansion culture The liquid is counted with a hemocytometer, and the number of bacteria in the secondary expansion culture liquid reaches 200-300 million/ml, preferably 200 million/ml.

The secondary liquid medium consists of: peptone 10.0g, beef extract 10.0g, agar 15.0g, molasses 5.0g, MgSO ₄ .7H ₂ O 0.2g, MnSO ₄ .H ₂ O 0.05g, CaCO ₃ 20.0g, NaCl 5.0 g, distilled water 1.0L. After mixing, sterilize at 112°C for 20 minutes, cool to room temperature naturally, and adjust the natural pH.

The function of the secondary culture is to make the expansion culture liquid and mix it to obtain the compound bacterial liquid.

4. According to the mass percentage content of the secondary expansion culture liquid of various strains: Bacillus subtilis 20-30%, Lactobacillus acidophilus 15-20%, Lactobacillus delbrueckii subspecies bulgaricus 15-20%, Saccharomyces cerevisiae 10-15%, Bacillus megaterium 10-15%, Clostridium pasteurianus 10-15%, the mass percentage content of each better strain is: Bacillus subtilis 25%, Lactobacillus acidophilus 15%, Germany 15% of Lactobacillus subsp. bulgaricus, 15% of Saccharomyces cerevisiae, 15% of Bacillus megaterium, and 15% of Clostridium pasteurianus are mixed to obtain a composite bacterial liquid. Adsorb the composite bacterial solution onto the natural zeolite powder with a particle size of 200 mesh, the mass ratio of the natural zeolite powder with a particle size of 200 mesh to the composite bacterial solution with the composite bacterial strain is 1-3:1, and then dry it in a low-temperature vacuum according to the existing technology to obtain a composite Microbial starter.

The strains used in the composite microbial starter of the present invention are selected for eliminating odor and harmful pathogenic bacteria produced by organic wastes, comprehensively utilizing organic wastes and improving the efficacy and quality of organic fertilizers. Bacillus subtilis can produce active antibacterial substances such as subtilisin, nystatin and lactic acid, which can inhibit the growth of other pathogenic bacteria; at the same time, it can produce a large amount of protease, lipase, amylase and glucoamylase, which can degrade organic waste such as straw , food waste, complex organic matter in sludge. Utilizing the characteristics of acid production and mellow aroma produced by Saccharomyces cerevisiae can reduce the odor produced in the fermentation process of organic waste such as poultry manure and kitchen waste. Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus acidophilus can produce a large amount of lactic acid, etc., which can inhibit the growth and reproduction of harmful microorganisms, kill harmful bacteria such as Escherichia coli and Salmonella, and produce a variety of vitamins and organic acids at the same time, which can promote crop growth. Bacillus megaterium has the function of dissolving phosphorus and potassium, which can be used by crops to absorb phosphorus and potassium. Clostridium pasteurianum has nitrogen fixation and supplies nitrogen for crops to absorb.

Example 1

1. Inoculation: Inoculate Bacillus subtilis, Lactobacillus acidophilus, Lactobacillus delbrueckii subspecies bulgaricus, Saccharomyces cerevisiae, Bacillus megaterium and Clostridium pasteurianus respectively, at room temperature, humidity is 50%, cultivate 18h, then Each was added into sterile water, shaken for 30 minutes to obtain a bacterial liquid, and the number of bacteria in the bacterial liquid reached 200 million/ml respectively.

Two, first-level culture, inoculate the strains of each bacterium solution respectively in a 50ml conical flask filled with a first-level liquid medium, the inoculum size is 5% v/v, the pH value of the medium is 6.00, and the conical flask Put it into a shaking table and shake it at 180r/min for 32h, and the number of bacteria in the bacterial solution after the first-level expansion reaches 200 million/ml.

3. Secondary culture, mix each bacterial solution of the first-level expansion culture with the second-level liquid medium and inoculate it into a 1000ml Erlenmeyer flask containing the second-level liquid medium, the inoculation amount is 5% v/v, conical The bottle was placed in a shaker and cultured by shaking at 180r/min for 48 hours to obtain a secondary expansion culture liquid, and the number of bacteria in the secondary expansion culture liquid reached 200 million/ml respectively.

4. According to the mass percentage content of the secondary expansion culture liquid of various strains: Bacillus subtilis 25%, Lactobacillus acidophilus 20%, Lactobacillus delbrueckii bulgaricus 20%, Saccharomyces cerevisiae 15%, Bacillus megaterium 10% %, Clostridium pasteuriani 10%, mixed to obtain a composite bacteria liquid. Adsorb 200 million bacteria per milliliter carrier onto the natural zeolite powder with a particle size of 200 mesh, the mass ratio of the natural zeolite powder with a particle size of 200 mesh to the composite bacteria is 1:1, and dry in vacuum at low temperature to obtain a composite microbial starter.

Example 2

1. Inoculation: Bacillus subtilis, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Saccharomyces cerevisiae, Bacillus megaterium and Clostridium pasteurianus were inoculated on a slant, at room temperature, the humidity was 50%, cultivated for 18 hours, Then each was added into sterile water, shaken for 30 minutes to obtain a bacterial liquid, and the number of bacteria in the bacterial liquid reached 200 million/ml respectively.

Two, primary culture, inoculate the bacterial classification of each bacterial liquid described in the 50ml Erlenmeyer flask that fills primary liquid culture medium respectively, inoculum size 8%v/v, the pH value of culture medium is 6.00, Erlenmeyer flask Put it into a shaking table and shake it at 180r/min for 32h, and the number of bacteria in the bacterial solution after the first-level expansion reaches 200 million/ml.

3. Secondary culture, mix each bacterial solution of the first-level expansion culture with the second-level liquid medium and inoculate it into a 1000ml Erlenmeyer flask containing the second-level liquid medium, the inoculation amount is 5% v/v, conical The bottle was placed in a shaker and cultured by shaking at 180r/min for 48 hours to obtain a secondary expansion culture liquid, and the number of bacteria in the secondary expansion culture liquid reached 200 million/ml respectively.

4. According to the mass percentage content of secondary expansion culture liquid of various strains: Bacillus subtilis 20%, Lactobacillus acidophilus 15%, Lactobacillus delbrueckii bulgaricus 20%, Saccharomyces cerevisiae 15%, Bacillus megaterium 15% %, Clostridium pasteurianus 15%, mixed to obtain a composite bacteria liquid. Adsorb 200 million bacteria per milliliter carrier onto the natural zeolite powder with a particle size of 200 mesh, the mass ratio of the natural zeolite powder with a particle size of 200 mesh to the composite bacteria is 1:1, and dry in vacuum at low temperature to obtain a composite microbial starter.

Example 3

1. Inoculation: Bacillus subtilis, Lactobacillus acidophilus, Lactobacillus delbrueckii subspecies bulgaricus, Saccharomyces cerevisiae, Bacillus megaterium and Clostridium pasteurianus were inoculated on a slant, at room temperature, humidity was 50%, cultivated for 20h, Then each was added into sterile water, shaken for 30 minutes to obtain a bacterial liquid, and the number of bacteria in the bacterial liquid reached 300 million/ml respectively.

Two, first-level culture, inoculate the strains of each bacterial solution in a 50ml conical flask filled with a first-level liquid medium respectively, the inoculum size is 10% v/v, the pH value of the medium is 6.00, and the conical flask Put it into a shaking table and shake it at 180r/min for 32h, and the number of bacteria in the bacterial solution after the first-level expansion reaches 300 million/ml.

3. Secondary culture, mix each bacterial solution of the first-level expansion culture with the second-level liquid medium and inoculate it into a 1000ml Erlenmeyer flask containing the second-level liquid medium, the inoculation amount is 10% v/v, conical The bottle was put into a shaker and cultured by shaking at 180r/min for 48 hours to obtain a secondary expansion culture liquid, and the number of bacteria in the secondary expansion culture liquid reached 300 million/ml respectively.

4. According to the mass percentage content of the secondary expansion culture liquid of various strains: Bacillus subtilis 20%, Lactobacillus acidophilus 20%, Lactobacillus delbrueckii bulgaricus 15%, Saccharomyces cerevisiae 15%, Bacillus megaterium 15% %, Clostridium pasteurianus 15%, mixed to obtain a composite bacteria liquid. Adsorb 300 million bacteria per milliliter carrier onto the natural zeolite powder with a particle size of 200 mesh, the mass ratio of the natural zeolite powder with a particle size of 200 mesh to the composite bacteria is 3:1, and dry in vacuum at low temperature to obtain a composite microbial starter.

Example 4

1. Inoculation: Bacillus subtilis, Lactobacillus acidophilus, Lactobacillus delbrueckii subspecies bulgaricus, Saccharomyces cerevisiae, Bacillus megaterium and Clostridium pasteurianus were inoculated on a slant, at room temperature, the humidity was 50%, cultivated for 16 hours, Then each was added into sterile water, shaken for 30 minutes to obtain a bacterial liquid, and the number of bacteria in the bacterial liquid reached 200 million/ml respectively.

Two, primary culture, inoculate the bacterial classification of each bacterial liquid described in the 50ml Erlenmeyer flask that fills primary liquid culture medium respectively, inoculum size 8%v/v, the pH value of culture medium is 6.00, Erlenmeyer flask Put it in a shaking table and shake it at 180r/min for 24 hours, and the number of bacteria in the bacterial solution after the first-level expansion reaches 200 million/ml.

3. Secondary culture, mix each bacterial solution of the first-level expansion culture with the second-level liquid medium and inoculate it into a 1000ml Erlenmeyer flask containing the second-level liquid medium, the inoculation amount is 8% v/v, conical The bottle was put into a shaker and cultured with 180r/min vibration for 72h to obtain the secondary expansion culture liquid, and the number of bacteria in the secondary expansion culture liquid reached 200 million/ml respectively.

4. According to the mass percentage content of the secondary expansion culture liquid of various strains: Bacillus subtilis 30%, Lactobacillus acidophilus 15%, Lactobacillus delbrueckii bulgaricus 15%, Saccharomyces cerevisiae 15%, Bacillus megaterium 10% %, Clostridium pasteurianus 15%, mixed to obtain a composite bacteria liquid. Adsorb 200 million bacteria per milliliter carrier onto the natural zeolite powder with a particle size of 200 mesh, the mass ratio of the natural zeolite powder with a particle size of 200 mesh to the composite bacteria is 3:1, and dry it in vacuum at low temperature to obtain a composite microbial starter.

Example 5

1. Inoculation: Bacillus subtilis, Lactobacillus acidophilus, Lactobacillus delbrueckii subspecies bulgaricus, Saccharomyces cerevisiae, Bacillus megaterium and Clostridium pasteurianus were inoculated on a slant, at room temperature, the humidity was 50%, cultivated for 16 hours, Then each was added into sterile water, shaken for 30 minutes to obtain a bacterial liquid, and the number of bacteria in the bacterial liquid reached 200 million/ml respectively.

Two, first-level culture, inoculate the strains of each bacterium solution respectively in a 50ml conical flask filled with a first-level liquid medium, the inoculum size is 5% v/v, the pH value of the medium is 6.00, and the conical flask Put it into a shaking table and shake it at 180r/min for 48 hours, and the number of bacteria in the bacterial solution after the first-level expansion reaches 200 million/ml.

3. Secondary culture, mix each bacterial solution of the first-level expansion culture with the second-level liquid medium and inoculate it into a 1000ml Erlenmeyer flask containing the second-level liquid medium, the inoculation amount is 5% v/v, conical The bottle was put into a shaker and cultured with 180r/min vibration for 72h to obtain the secondary expansion culture liquid, and the number of bacteria in the secondary expansion culture liquid reached 200 million/ml respectively.

4. According to the mass percentage content of secondary expansion culture liquid of various strains: Bacillus subtilis 30%, Lactobacillus acidophilus 20%, Lactobacillus delbrueckii bulgaricus 15%, Saccharomyces cerevisiae 10%, Bacillus megaterium 15% %, Clostridium pasteuriani 10%, mixed to obtain a composite bacteria liquid. Adsorb 200 million bacteria per milliliter carrier onto the natural zeolite powder with a particle size of 200 mesh, the mass ratio of the natural zeolite powder with a particle size of 200 mesh to the composite bacteria is 1:1, and dry in vacuum at low temperature to obtain a composite microbial starter.

Application example 1

Adopt the compound microbial starter of embodiment 1, ferment decomposed main material A, main material B and auxiliary material, make powdery bio-organic fertilizer. The raw material of the main ingredient A is wet chicken manure with a mass solid content of not less than 70%, the raw material of the main ingredient B is sewage sludge with a mass solid content of not less than 70%, and the raw material of the auxiliary material is dry straw.

The preparation method is as follows:

1. Crush the straw to no longer than 10mm to make auxiliary materials.

2. Mix 700 kg of wet chicken manure, 300 kg of auxiliary materials and 2 kg of compound microbial starter, mix evenly, pile up 1-2 meters high, turn the pile once after 15 days, stack and decompose for 30 days, and make main ingredient A.

Mix 700 kg of domestic sewage sludge, 300 kg of auxiliary materials and 2 kg of compound microbial starter, mix evenly, pile up to a height of 1-2 meters, turn the pile once after 15 days, and pile up and decompose for 30 days to make the main ingredient B.

3. Mix 500 kg of main ingredient A, 500 kg of main ingredient B, 50 kg of ammonium sulfate, 50 kg of superphosphate, and 50 kg of aluminum magnesium silicate powder, and then ferment. During the fermentation, keep the temperature at 45-65°C. When the temperature exceeds 65°C, use a compost turner to turn the compost to cool down, so as to prevent the fermented material from agglomerating and make it evenly mixed. The compost is stacked and decomposed for 25 days to complete the fermentation.

4. Crush the fermented material to break up the agglomerated compost, and then sieve to further remove impurities.

5. Measure and pack in bags to obtain powdery bio-organic fertilizer.

Tested according to NY884-2012 "Biological Organic Fertilizer" standard, its effective components have organic matter ≥ 40%, the number of effective viable bacteria cfu ≥ 0.2 billion/g, the number of coliforms ≤ 100/g (mL), and the death rate of roundworm eggs ≥ 95%, moisture content ≤ 30%, no odor.

Application example 2

The composite microbial starter of Example 2 was used to ferment the decomposed main material A, main material B and auxiliary materials to make powdery bio-organic fertilizer. The raw material of main ingredient A is wet pig manure with a mass solid content of not less than 70%, the raw material of main ingredient B is wet chicken manure with a mass solid content of not less than 70%, and the raw material of auxiliary materials is pig manure with a mass water content of not more than 35%. Mushroom dregs.

The preparation method is as follows:

1. Crush mushroom dregs to a size no larger than 10mm to make auxiliary materials.

2. Mix 700 kg of wet pig manure, 300 kg of auxiliary materials and 2 kg of compound microbial starter, mix evenly, pile up 1-2 meters high, turn the pile once after 15 days, stack and decompose for 30 days, and make main ingredient A.

700 kg of wet chicken manure, 300 kg of auxiliary materials and 2 kg of compound microbial starter are evenly spread and mixed, piled up to a height of 1-2 meters, turned once after 15 days, piled and decomposed for 30 days, and made into main ingredient B.

3. Mix 500 kg of main ingredient A, 500 kg of main ingredient B, 50 kg of ammonium sulfate, 50 kg of superphosphate, and 50 kg of aluminum magnesium silicate powder, mix them evenly, and carry out fermentation. During the fermentation period, keep the temperature at 45-65°C. When the fermentation temperature exceeds 65°C, use a compost turner to turn the compost to cool down, so as to prevent the fermented material from agglomerating and make it evenly mixed, and stack and decompose for 15 days.

4. Crush the fermented material to break up the agglomerated compost, and then sieve to further remove impurities.

5. Measure and pack in bags to obtain powdery bio-organic fertilizer.

Tested according to NY884-2012 "Biological Organic Fertilizer" standard, its effective components have organic matter ≥ 40%, the number of effective viable bacteria cfu ≥ 0.2 billion/g, the number of fecal coliforms ≤ 100/g (mL), and the mortality rate of roundworm eggs ≥95%, moisture content ≤30%, no odor.

Application example 3

The composite microbial starter of Example 3 was used to ferment the decomposed main material A, main material B and auxiliary materials to make powdery bio-organic fertilizer. The raw material of the main ingredient A is cow dung with a mass solid content of not less than 70%, the raw material of the main ingredient B is sheep manure with a mass solid content of not less than 70%, and the raw material of the auxiliary material is dry cotton stalk.

The preparation method is as follows:

1. Crush the cotton stalks to a size no larger than 10mm to make auxiliary materials.

2. Mix 700 kg of cow dung, 300 kg of auxiliary materials and 2 kg of compound microbial starter, mix evenly, pile up 1-2 meters high, turn the pile once after 15 days, stack and decompose for 30 days, and make the main ingredient A.

Mix 700 kg of sheep manure, 300 kg of auxiliary materials and 2 kg of compound microbial starter, mix evenly, pile up a height of 1-2 meters, turn the pile once after 15 days, pile up and decompose for 30 days, and make the main ingredient B.

3. Mix 500 kg of main ingredient A, 500 kg of main ingredient B, 50 kg of ammonium sulfate, 50 kg of superphosphate, and 50 kg of aluminum magnesium silicate powder, mix them evenly, and carry out fermentation. During the fermentation period, keep the temperature at 45-65°C. When the fermentation temperature exceeds 65°C, use a compost turner to turn the compost to cool down, so as to prevent the fermented material from agglomerating and make it evenly mixed, and stack and decompose for 15 days.

4. Crush the fermented material to break up the agglomerated compost, and then sieve to further remove impurities.

5. Measure and pack in bags to obtain powdery bio-organic fertilizer.

Tested according to NY884-2012 "Biological Organic Fertilizer" standard, its effective components have organic matter ≥ 40%, the number of effective viable bacteria cfu ≥ 0.2 billion/g, the number of fecal coliforms ≤ 100/g (mL), and the mortality rate of roundworm eggs ≥95%, moisture content ≤30%, no odor.

Claims (10)

1. a compound microbial culture starter, it is characterized in that: described compound microbial culture starter is made up of carrier and composite bacteria, mix with the mass ratio 1-3:1 of the composite bacteria liquid of composite bacteria by carrier, obtain after drying, described carrier is granularity 200 object natural zeolite powder, described composite bacteria liquid is made up of the mass percent of following bacterial classification bacterium liquid: subtilis 20-30%, Lactobacterium acidophilum 15-20%, lactobacillus delbruockii subspecies bulgaricus 15-20%, yeast saccharomyces cerevisiae 10-15%, bacillus megaterium 10-15%, Pasteur's Clostridium 10-15%, described each bacterial classification bacterium liquid every ml of carrier 2-3 hundred million bacterium number.

2. compound microbial culture starter according to claim 1, it is characterized in that: described composite bacteria liquid is made up of the mass percent of following bacterium liquid: subtilis 25%, Lactobacterium acidophilum 15%, lactobacillus delbruockii subspecies bulgaricus 15%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 15%, Pasteur's Clostridium 15%, described each bacterium liquid every ml of carrier 200,000,000 bacterium number.

3. a preparation method for compound microbial culture starter, comprises the following steps:

One, inoculate, respectively by subtilis, Lactobacterium acidophilum, lactobacillus delbruockii subspecies bulgaricus, yeast saccharomyces cerevisiae, bacillus megaterium and Pasteur's Clostridium, under room temperature, relative humidity is 50%, cultivate 16-20h, then respectively join in sterilized water, vibrate and obtain bacterium liquid after 30 minutes;

Two, one-level is cultivated, described each bacterium liquid is inoculated in the Erlenmeyer flask filling level liquid substratum respectively, inoculum size is 5%-10%, the pH value of substratum is 6.0-8.0,24-48h is cultivated with 180r/min rotating speed, obtain one-level to spread cultivation bacterium liquid, the one-level bacterium quantity in bacterium liquid that spreads cultivation reaches 2-3 hundred million/ml respectively;

Three, secondary is cultivated, each one-level being spread cultivation after bacterium liquid mixes with secondary liquid substratum is inoculated in the Erlenmeyer flask being loaded with secondary liquid substratum, inoculum size is 5%-10%, 48-72h is cultivated with 180r/min rotating speed, obtain secondary to spread cultivation bacterium liquid, the secondary bacterium quantity in bacterium liquid that spreads cultivation reaches 2-3 hundred million/ml respectively;

Four, by the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary be: subtilis 20-30%, Lactobacterium acidophilum 15-20%, lactobacillus delbruockii subspecies bulgaricus 15-20%, yeast saccharomyces cerevisiae 10-15%, bacillus megaterium 10-15%, Pasteur's Clostridium 10-15%, is mixed to get composite bacteria liquid; Composite bacteria liquid is adsorbed onto on granularity 200 object natural zeolite powder, granularity 200 object natural zeolite powder with have the mass ratio of the composite bacteria liquid of composite bacteria to be 1-3:1, dry, obtain compound microbial culture starter.

4. the preparation method of compound microbial culture starter according to claim 3, is characterized in that: described step 2 inoculum size is 10%, and the pH value of described substratum is 6.0-7.0; Described with 180r/min rotating speed, cultivate 32h.

5. the preparation method of compound microbial culture starter according to claim 4, is characterized in that: bacterium quantity 200,000,000/ml respectively in described step 2 bacterium liquid.

6. the preparation method of compound microbial culture starter according to claim 5, is characterized in that: the level liquid substratum composition of described step 2 is respectively:

(1) subtilis level liquid substratum consists of: peptone 5.0g, beef leaching thing 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to room temperature, pH7.0;

(2) Lactobacterium acidophilum level liquid substratum consists of: casein peptone 10.0g, extractum carnis 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, sorbitan monooleate Soxylat A 25-7 1.0g, K ₂hPO ₄2.0g, MgSO ₄.7H ₂o 0.2g, MnSO ₄.H2O 0.05g, CaCO ₃20.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to 37 DEG C, pH6.8;

(3) lactobacillus delbruockii subspecies bulgaricus level liquid substratum consists of: casein peptone 10.0g, extractum carnis 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, Tween 801.0g, K ₂hPO ₄2.0g, MgSO ₄.7H ₂o 0.2g, MnSO ₄.H ₂o 0.05g, CaCO ₃20.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to 37 DEG C, pH6.8;

(4) yeast saccharomyces cerevisiae level liquid substratum consists of: 5 ° of B é wort 1.0L, agar 15.0g; Mix rear 112 DEG C of sterilizing 20min, natural in 28 DEG C, natural pH;

(5) bacillus megaterium level liquid substratum consists of: peptone 5.0g, beef leaching thing 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to 30 DEG C, pH7.0;

(6) Pasteur's Clostridium level liquid substratum consists of: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to culture temperature 37 DEG C, pH7.0.

7. the preparation method of compound microbial culture starter according to claim 6, is characterized in that: described step 3 inoculum size is 10%; Described with 180r/min rotating speed cultivation 48h.

8. the preparation method of compound microbial culture starter according to claim 7, is characterized in that: the secondary of the described step 3 bacterium quantity in bacterium liquid that spreads cultivation is respectively 200,000,000/ml.

9. the preparation method of compound microbial culture starter according to claim 8, is characterized in that: the secondary liquid substratum of described step 3 consists of: peptone 10.0g, extractum carnis 10.0g, agar 15.0g, molasses 5.0g, MgSO ₄.7H ₂o 0.2g, MnSO ₄.H ₂o 0.05g, CaCO ₃20.0g, NaCl 5.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to room temperature, natural pH.

10. the preparation method of compound microbial culture starter according to claim 9, it is characterized in that: the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary of described step 4 is: subtilis 25%, Lactobacterium acidophilum 15%, lactobacillus delbruockii subspecies bulgaricus 15%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 15%, Pasteur's Clostridium 15%.