Preparation method of humic acid type straw source microbial agent

Abstract

The invention discloses a humic acid type straw source microbial agent which comprises the following strains: enterococcus faecium, bacillus subtilis, lactobacillus plantarum and saccharomyces cerevisiae. The method has the advantages that turfy soil, biogas residues, biogas slurry and a certain amount of agricultural waste are added into compost with straw microbial agents and livestock and poultry manure as main materials to adjust a proper carbon nitrogen ratio, and more importantly, a compound microbial agent for strengthening the process of the humic acid type straw source microbial agent is inoculated; compared with natural composting fermentation, the composting efficiency and stability are improved, and the fertilizer efficiency and fertility of the organic fertilizer and the resource utilization efficiency of waste are improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological agents, in particular to a preparation method of a humic acid type straw source microbial agent. BACKGROUND

[0002] China is a large agricultural country, and grain production provides an important foundation and guarantee for the people's livelihood, but the disposal of agricultural waste has become a problem. At present, straw is mainly disposed of by burning, which not only causes serious waste of resources, but also causes serious environmental pollution problems, damages human health, and in addition, the high temperature generated during the burning process also destroys the flora and organic matter in the soil, reducing the fertility of the soil, which is extremely harmful. If the straw can be reasonably utilized and used as a raw material for organic fertilizer fermentation, it not only reduces haze, but also solves the problem of livestock and poultry manure disposal, and at the same time, the well-prepared organic fertilizer applied to the cultivated land can also regulate the flora structure, increase the organic matter and various mineral elements, which can not only reduce the use of chemical fertilizers, but also repair the land damaged by pesticides and fertilizers, which is a multiple effect. SUMMARY

[0003] The present application aims to provide a preparation method of a humic acid type straw source microbial agent to solve the problems in the background art.

[0004] To achieve the above-mentioned purpose, the present application provides the following technical scheme: a humic acid type straw source microbial agent, characterized in that it comprises the following strains: Enterococcus faecium, Bacillus subtilis, Lactobacillus plantarum and Saccharomyces cerevisiae.

[0005] 2. The preparation method of the humic acid type straw source microbial agent according to claim 1, wherein the proportion of the strains is: total number of Bacillus subtilis : total number of Enterococcus faecium : total number of Lactobacillus plantarum : total number of Bacillus licheniformis : total number of Saccharomyces cerevisiae = 7:10:2.5:3.5:1.

[0006] 3. A preparation method of a humic acid type straw source microbial agent, characterized in that it comprises the following steps:

[0007] Step 1: strain culture: inoculate Enterococcus faecium, Bacillus subtilis WE, Bacillus licheniformis, Lactobacillus plantarum and Saccharomyces cerevisiae into respective slant culture medium, wherein Enterococcus faecium and Lactobacillus plantarum use agar medium, 37℃ culture for 24h, Bacillus subtilis and Bacillus licheniformis use agar medium, 37℃ culture for 24h, and Saccharomyces cerevisiae uses YPD agar medium, 30℃ culture for 48h.

[0008] 4. A preparation method of humic acid type straw source microbial agent, wherein each strain is separately cultured in a 500 mL shake flask containing 200 mL seed medium, Enterococcus faecium and Lactobacillus plantarum use MRS liquid medium, 37°C static culture for 20 hours, Bacillus subtilis and Bacillus licheniformis use LB liquid medium, 37°C, 180 rpm culture for 20 hours, Saccharomyces cerevisiae uses liquid medium, 30°C, 180 rpm culture for 40 hours, then the bacterial liquid is mixed in equal volume to prepare 1 L seed liquid.

[0009] 5. The preparation method of humic acid type straw source microbial agent according to claim 4, wherein the seed liquid is all inoculated into a 200 L seed tank containing 160 L fermentation medium, 37°C, 100 rpm culture for 20 hours, then all inoculated into a 5 T fermentation tank containing 3 T fermentation medium, 37°C, 100 rpm culture for 40 hours.

[0010] 6. The preparation method of humic acid type straw source microbial agent according to claim 1, wherein the fermentation medium comprises 22 g of glucose, 5.5 g of yeast powder, 30 g of corn syrup dry powder, 0.4 g of MnSO4·H2O, 0.4 g of MgSO4·7H2O, 0.8 g of K2HPO4·3H2O, 800 mL of water, and the pH is adjusted to 7.0, and sterilized at 121°C for 15 minutes.

[0011] 7. The application also provides an organic fertilizer application of humic acid type straw source microbial agent, wherein the whole fermentation process is implemented on site in a breeding farm.

[0012] 8. An application method of humic acid type straw source microbial agent, comprising the following steps: mixing animal manure with auxiliary materials at a weight ratio of 1:2, adjusting the carbon-nitrogen ratio to 25-30:1, inoculating the compound microbial agent at a weight ratio of 1-4%, adding water and biogas fermentation liquid to adjust the water content to 50-65%, regularly monitoring the temperature and water content, and adding water to maintain the compost water content at 55-65%, turning the compost when the temperature significantly decreases, turning the compost for 2-3 times, and preparing the organic fertilizer after the temperature no longer significantly increases and the composting for 5-10 days.

[0013] Beneficial effects

[0014] The preparation method of humic acid type straw source microbial agent provided by the application adds grass charcoal, biogas residue, biogas liquid and a certain amount of agricultural waste to the compost mainly composed of straw microbial agent and livestock and poultry manure to adjust the appropriate carbon-nitrogen ratio, and more importantly, inoculates the compound microbial agent for strengthening the process of humic acid type straw source microbial agent, compared with natural compost fermentation, the efficiency and stability of the compost are improved, and the fertilizer efficiency and fertility of the organic fertilizer and the resource utilization efficiency of the waste are improved. DETAILED DESCRIPTION

[0015] The following are specific embodiments of the present application, which further describe the technical solutions of the present application, but the present application is not limited to these embodiments.

[0016] Example 1

[0017] The strains in the composite microbial agent are isolated from naturally fermented straws. The isolated strains are subjected to experiments of single strain, two-strain combination and multi-strain complex on the fermentation effect of organic fertilizer for multiple times, and strains capable of significantly improving the composting efficiency are screened. Then, the safety screening experiments of bacteria such as endotoxin, hemolysis, cytotoxicity and biogenic amine production are carried out. Finally, the composition of the straw organic fertilizer fermentation composite microbial agent is determined as Enterococcus faecium WEI-6, Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum and Saccharomyces cerevisiae.

[0018] Strain culture: Enterococcus faecium, Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum and Saccharomyces cerevisiae are inoculated into their respective slant media. Enterococcus faecium and Lactobacillus plantarum are cultured at 37°C for 24h using MRS agar medium, Bacillus subtilis and Bacillus licheniformis are cultured at 37°C for 24h using LB agar medium, and Saccharomyces cerevisiae is cultured at 30°C for 48h using YPD agar medium. Preparation of composite microbial agent: each strain is separately cultured in a 500mL shake flask containing 200mL seed medium. Enterococcus faecium WEI-6 and Lactobacillus plantarum are cultured at 37°C for 20h using MRS liquid medium, Bacillus subtilis and Bacillus licheniformis are cultured at 37°C and 180rpm for 20h using LB liquid medium, and Saccharomyces cerevisiae WEI-92 is cultured at 30°C and 180rpm for 40h using YPD liquid medium. Then, the bacterial liquids are mixed in equal volumes to prepare 1L seed liquid. The seed liquid is inoculated into a 200L seed tank containing 160L fermentation medium, and cultured at 37°C and 100rpm for 20h. Then, the whole is inoculated into a 5T fermentation tank containing 3T fermentation medium, and cultured at 37°C and 100rpm for 40h.

[0019] The mixed fermentation medium used in the above scheme is as follows:

[0020] Glucose 20g, yeast powder 5g, corn syrup dry powder 25g, MnSO4·H2O 0.2g, MgSO4·7H2O 0.2g, K2HPO4·3H2O 1g, water 1000mL, pH 7.0, sterilized at 121°C for 20min.

[0021] The composite microbial agent is characterized in that the proportion of different strains is as follows: total number of Bacillus subtilis : total number of Enterococcus faecium : total number of Lactobacillus plantarum : total number of Bacillus licheniformis : total number of Saccharomyces cerevisiae = 4:10:2:3:1. The total number of lactic acid bacteria, the total number of yeasts and the total number of spores are shown in Table 1.

[0022] Example Two

[0023] Operation process of on-site fermentation of organic fertilizer by straw in large-scale pig farms

[0024] The composite microbial agent prepared in Example One is used for on-site fermentation to produce organic fertilizer in large-scale pig farms. The on-site fermentation preparation method comprises the following steps:

[0025] Preparation of fermentation pool: the fermentation pool is a soil pit with a length of 50 meters, a width of 30 meters and a depth of 2 meters. The straw is packaged into bundles by a baling machine, each bundle weighing about 600 kg, and a total of 400 T of straw is transported to the fermentation pool and regularly placed on the north and south sides. The straw is watered regularly to maintain a certain humidity. A passageway is left in the middle of the fermentation pool for the convenience of vehicles to enter and exit. A manure storage tank is prepared beside the fermentation pool for accumulation of pig manure.

[0026] Operation process of organic fertilizer

[0027] Manure treatment: 100 T of pig manure is transported to the top of 150 T of straw by a forklift and is evenly spread as much as possible. Other auxiliary materials are added: 30 T of grass carbon soil, 20 T of biogas residue and household garbage and agricultural waste (rice straw, fruit shells, dead pigs, etc.) are added at the same time. The animal manure and the auxiliary materials are mixed evenly at a weight ratio of 1:2.

[0028] Addition of microbial agent: 3 T of mixed microbial agent is mixed with 5 T of biogas slurry and appropriate water, and then all the microbial agents are evenly sprayed onto the pig manure and straw by a water pump so that the inoculation amount reaches 1-5% by weight. After mixing, water and biogas fermentation liquid are added to adjust the moisture content to 55-65%.

[0029] Fermentation: start fermentation, turn the pile when the temperature drops significantly, turn the pile 2-3 times in total, and then let it mature for 5-10 days to obtain the organic fertilizer.

[0030] Operation process of organic fertilizer,

[0031] Manure treatment: 100 T of pig manure and 50 T of other auxiliary materials are transported to the top of 250 T of straw by a forklift and are evenly spread as much as possible. Other auxiliary materials are added: 30 T of grass carbon soil, 20 T of biogas residue and household garbage and agricultural waste (rice straw, fruit shells, dead pigs, etc.) are added at the same time. The animal manure and the auxiliary materials are mixed evenly at a weight ratio of 1:3.

[0032] Adding bacteria: 4T mixed bacteria and 5T biogas liquid and appropriate water are mixed, and all bacteria are uniformly sprayed on pig manure and straw by water pump, so that the amount is 1-5% by weight, and then water and biogas fermentation liquid are added to adjust the moisture content to 55-65%;

[0033] Fermentation: Start fermentation, turn over when the temperature drops significantly, turn over 2-3 times in total, and then mature for 5-10 days to obtain organic fertilizer. The control operation process: use compost without adding compound bacteria as the control group.

[0034] Example three

[0035] Process monitoring of scale pig farm using straw on-site fermented organic fertilizer

[0036] Organic fertilizer: On the second day after the start of fermentation, sporadic hot spots began to appear, and the temperature was detected to be above 60°C. In places without obvious heat, the temperature remained around 30°C. After that, the hot spots gradually increased, and by the third day, a large number of hot spots began to connect into patches, with a central temperature of around 70°C. Water was added to the straw every 2 days. On the fifth day, the temperature began to drop significantly, and the entire pile was thoroughly mixed again using an excavator. On the second day, a large amount of heat began to generate, and on the eighth day, the pile was turned over again. The high temperature lasted for 2 days, and then entered the maturation stage, which lasted for about 5-10 days. The entire organic fertilizer fermentation process took about 15-20 days, and the product could be field scattered or bagged.

[0037] Control group: On the fifth day of fermentation, sporadic hot spots began to appear, with a temperature of above 60°C. By the eighth day, the hot spots connected into patches, and the central temperature remained between 60°C and 70°C. Water was added every 3 days. On the eleventh day, the temperature began to drop significantly, and the pile was turned over using an excavator. The temperature did not rise, and then entered the maturation stage, which lasted for about 10-15 days. The fermentation process was completed. It can be seen that the control group did not experience a second temperature rise, and the fermentation of straw and manure was far from complete compared to the experimental group.

[0038] Organic fertilizer: On the third day after the start of fermentation, sporadic hot spots began to appear, and the temperature was detected to be above 60°C. In places without obvious heat, the temperature remained around 30°C. After that, the hot spots gradually increased, and by the fourth day, a large number of hot spots began to connect into patches, with a central temperature of around 70°C. Water was added to the straw every 1 day. On the seventh day, the temperature began to drop significantly, and the entire pile was thoroughly mixed again using an excavator. On the second day, a large amount of heat began to generate, and on the tenth day, the pile was turned over again. The high temperature lasted for 2 days, and then entered the maturation stage, which lasted for about 5-10 days. The entire organic fertilizer fermentation process took about 17-22 days, and the product could be field scattered or bagged.

[0039] Control group: On the seventh day of fermentation, sporadic hot spots began to appear, with a temperature of above 60°C. Water was added every 3 days.

[0040] On the 11th day, the compost was turned over once with an excavator, increasing the number of hot spots, but large-scale heating never occurred. The fermentation process ended after about 15 days. It is evident that the heating in the control group was consistently unsatisfactory, and the fermentation of straw and manure was far less thorough than in the experimental group.

[0041] Example 4

[0042] Operational process of using straw to ferment organic fertilizer on-site in large-scale chicken farms

[0043] The compound microbial agent prepared in Example 1 was used for on-site fermentation production of organic fertilizer in a large-scale chicken farm. The on-site fermentation preparation method included the following steps: Preparation of the fermentation tank: The fermentation tank was an earthen pit 60 meters long, 40 meters wide, and 2 meters deep. Straw was bundled into bales using a baler, each bale weighing approximately 600 kg, totaling 900 tons of straw. These bales were transported to the fermentation tank, and the straw was regularly watered to maintain a certain level of humidity. [The text abruptly ends here, likely due to an incomplete sentence or missing information.]

[0044] Prepare a septic tank.

[0045] Manure treatment: In the manure storage tank, add 100 tons of chicken manure to approximately 100 tons of domestic sewage. After mixing with a loader, transport it to the top of 450 tons of straw and spread it as evenly as possible. Add other auxiliary materials: Simultaneously add 20 tons of peat moss, 30 tons of biogas slurry, and domestic waste and agricultural waste (rice straw, eggshells, dead chickens, etc.). Mix the animal manure and auxiliary materials evenly at a weight ratio of 1:5. Add microbial agents: Mix 10 tons of mixed microbial agents with 10 tons of biogas slurry and appropriate amount of water, and pump the mixture in two batches using a water pump to add all the microbial agents.

[0046] The agent is evenly sprayed onto the straw, so that the seed amount reaches 1-5% by weight. After mixing, water and biogas fermentation liquid are added to adjust the moisture content to 55-65%.

[0047] Fermentation: Start fermentation. When the temperature drops significantly, turn the pile over. Turn the pile over 2-3 times. After the temperature no longer rises significantly, let it decompose for 5-10 days to obtain organic fertilizer.

[0048] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A humic acid-based straw-derived microbial inoculant, characterized in that, It contains the following strains: Enterococcus faecalis, Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae.

2. The method for preparing humic acid-based straw-derived microbial inoculant according to claim 1, characterized in that, The strain composition ratio is as follows: total number of Bacillus subtilis: total number of Enterococcus faecalis: total number of Lactobacillus plantarum: total number of Bacillus licheniformis: total number of Saccharomyces cerevisiae = 7:10:2.5:3.5:

1.

3. A method for preparing a humic acid-based straw-derived microbial inoculant, characterized in that, Includes the following steps: Step 1: Strains culture: Enterococcus faecalis, Bacillus subtilis WE, Bacillus licheniformis, Lactobacillus plantarum, and Saccharomyces cerevisiae were inoculated into their respective slant agar media. Enterococcus faecalis and Lactobacillus plantarum were cultured on agar medium at 37°C for 24 hours. Bacillus subtilis and Bacillus licheniformis were cultured on agar medium at 37°C for 24 hours. Saccharomyces cerevisiae was cultured on YPD agar medium at 30°C for 48 hours.

4. A method for preparing a humic acid-based straw-derived microbial inoculant: Each strain of bacteria is first cultured separately in a 500mL shake flask containing 200mL of seed culture medium. Enterococcus faecalis and Lactobacillus plantarum are cultured on MRS liquid medium and incubated at 37℃ for 20h. Bacillus subtilis and Bacillus licheniformis are cultured on LB liquid medium and incubated at 37℃ and 180rpm for 20h. Saccharomyces cerevisiae is cultured on liquid medium and incubated at 30℃ and 180rpm for 40h. Then, the bacterial cultures are mixed in equal volumes to prepare 1L of seed culture.

5. The method for preparing humic acid-based straw-derived microbial inoculant according to claim 4, characterized in that, All the seed culture was inoculated into a 200L seed tank containing 160L of fermentation medium and cultured at 37℃ and 100rpm for 20h. Then, it was all inoculated into a 5T fermenter containing 3T of fermentation medium and cultured at 37℃ and 100rpm for 40h.

6. The method for preparing humic acid-based straw-derived microbial inoculant according to claim 1, characterized in that, The fermentation medium consists of: 22g glucose, 5.5g yeast powder, 30g corn steep liquor powder, 0.4g MnSO4·H2O, 0.4g MgSO4·7H2O, 0.8g K2HPO4·3H2O, and 800mL water. The pH is adjusted to 7.0, and the medium is sterilized at 121℃ for 15min.

7. This invention also provides an application of humic acid-based straw-derived microbial inoculants in organic fertilizer, characterized in that, The entire fermentation process was carried out on-site at the farm.

8. A method for applying a humic acid-based straw-derived microbial inoculant, characterized in that, The process includes the following steps: Mix animal manure and auxiliary materials at a weight ratio of 1:2, adjust the carbon-nitrogen ratio to 25-30:1, and simultaneously introduce compound microbial inoculants at a weight ratio of 1-4%. Add water and biogas fermentation liquid to adjust the moisture content to 50-65%. Regularly monitor the temperature and moisture content, and add water to maintain the compost moisture content at 55-65%. When the temperature drops significantly, turn the compost pile. Turn the pile 2-3 times in total. After the temperature no longer rises significantly, allow it to decompose for 5-10 days to obtain organic fertilizer.