CN112048449A - Compost composite microbial inoculum and application thereof - Google Patents
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Abstract
The invention discloses a compost composite microbial inoculum and application thereof, wherein the composite microbial inoculum comprises bacillus subtilis, bacillus licheniformis, trichoderma viride, aspergillus niger and saccharomycetes, and the bacillus subtilis: b, bacillus licheniformis: and (3) trichoderma viride: aspergillus niger: 0.2-0.8% of yeast: 0.2-0.8: 0.8-1.2: 0.8-1.2: 0.8 to 2.2. The screened compost composite bacterial agent has synergistic effect among strains, and can fully play the role of each strain in compost. The effect of the composite microbial inoculum inoculated and screened in the compost is mainly shown as follows: the temperature rise is accelerated, and the high temperature duration is prolonged; accelerating the decomposition of organic matters; nitrogen loss is reduced, and nutrient content is improved; the decomposition degree of the compost is improved, and the decomposition period is shortened; improving the fertilizer efficiency of compost products and the like.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a compost composite microbial inoculum and application thereof.
Background
The high-temperature aerobic composting is an effective way for realizing harmless and resource utilization of livestock and poultry manure at present, the microbial activity can be improved, the fermentation period can be shortened, the quality of compost products can be improved by artificially adding the microbial agent, and the high-temperature aerobic composting is widely applied to commercial composting. However, the loss of nitrogen is serious in the high-temperature composting process of the livestock and poultry manure, and researches show that the loss of nitrogen ranges from 16% to 74%, and ammonia is the most volatile. The microbial agent is added into the livestock manure compost material, so that the nitrogen loss can be reduced, the decomposition and conversion of organic matters are promoted, and the fermentation process of the livestock manure is accelerated. The conversion of nitrogen in the composting process is related to the material property, temperature, pH value and microbial species and distribution change in the composting, and the research on the dynamic change of nitrogen and the influence mechanism of microbes in the composting process under the condition of adding microbial agents is lacked at present. In different stages of composting (a temperature rise period, a high temperature period and a decomposition period), dominant populations with different properties exist, and the dominant populations influence the nitrogen change and the decomposition process of the composting process through interaction effects. The commercial microbial inoculum has a plurality of microbial varieties, but the effect is not ideal due to the property difference of materials, the antagonistic action of flora and the like. At present, compost microbial inoculum develops rapidly at home and abroad, but most of the compost microbial inoculum has no particularly ideal effect on nitrogen preservation. Most of the strains added in the compost are screened out by dominant strains in the fermented compost, and external functional strains can also be added. Species of microorganisms that may be used as foreign strains to add to the compost include bacteria: lactobacillus, Bacillus, Pseudomonas, Azotobacter, Microbacterium, etc.; fungi: trichoderma, Aspergillus, white rot fungi, Schizosaccharomyces; actinomycetes: streptomyces and Thermoactinomyces. However, screening a complex microbial inoculum to enable each strain to mutually cooperate to generate synergistic effect and fully exert the effect of each strain in compost is a technical problem to be solved urgently by technical personnel in the field. The invention tests that the dominant strains screened by the earlier stage natural compost are compounded and compared with the commercial microbial inoculum and the natural compost, researches the nitrogen dynamic change, the transformation rule and the microbial action mechanism in each composting stage under the condition of adding the microbial inoculum, discusses the reasons of the difference of different microbial inoculum proportions in reducing the nitrogen loss and promoting the maturity, and provides a certain scientific basis for optimizing the microbial inoculum with the functions of nitrogen preservation and decay promotion in the production of commercial organic fertilizers, optimizing the formula of the fecal compost microbial inoculum and reducing the nitrogen loss in the composting process.
Disclosure of Invention
The invention aims to provide a compost composite microbial inoculum and application thereof. The effect of the composite microbial inoculum inoculated and screened in the compost is mainly shown as follows: the temperature rise is accelerated, and the high temperature duration is prolonged; accelerating the decomposition of organic matters; nitrogen loss is reduced, and nutrient content is improved; the decomposition degree of the compost is improved, and the decomposition period is shortened; improving the fertilizer efficiency of compost products and the like.
The purpose of the invention can be realized by the following technical scheme:
a compost composite microbial inoculum comprises bacillus subtilis, bacillus licheniformis, trichoderma viride, aspergillus niger and saccharomycetes, wherein the bacillus subtilis comprises the following components in percentage by mass: b, bacillus licheniformis: and (3) trichoderma viride: aspergillus niger: 0.2-0.8% of yeast: 0.2-0.8: 0.8-1.2: 0.8-1.2: 1.8 to 2.2. As a preferred technical scheme, the bacillus subtilis comprises the following components in percentage by mass: b, bacillus licheniformis: and (3) trichoderma viride: aspergillus niger: yeast 0.5:0.5:1:1: 2.
Further preferably, the effective viable count of the bacillus subtilis is 200 hundred million/g; the number of effective live bacteria of the bacillus licheniformis is 200 hundred million/g, the number of effective live bacteria of the trichoderma viride is 100 hundred million/g, and the number of effective live bacteria of the aspergillus niger is 100 hundred million/g; the effective viable count of the yeast is 100 hundred million/g.
The composite microbial inoculum is applied to compost.
In the application, the addition amount of the composite microbial inoculum in the compost is 2-6 per mill by mass ratio. Preferably, the addition amount of the composite microbial inoculum in the compost is 4 per mill.
5 strains in the composite microbial inoculum are conventional commercial strains, and the compost composite microbial inoculum can be prepared by mixing the conventional commercial strains according to a proportion.
The invention has the beneficial effects that:
the screened compost composite bacterial agent has synergistic effect among strains, and can fully play the role of each strain in compost. The effect of the composite microbial inoculum inoculated and screened in the compost is mainly shown as follows: the temperature rise is accelerated, and the high temperature duration is prolonged; accelerating the decomposition of organic matters; nitrogen loss is reduced, and nutrient content is improved; the decomposition degree of the compost is improved, and the decomposition period is shortened; improving the fertilizer efficiency of compost products and the like.
Drawings
FIG. 1 is a graph of the average temperature of the upper, middle, lower and lower parts of a fungus chaff and commercial microbial inoculum group.
FIG. 2 is a graph of the average temperature of the upper, middle, lower and lower parts of the fungus chaff + mixed bacteria group.
FIG. 3 is a graph showing the change of water content in two sets.
FIG. 4 is a graph of two sets of pH changes.
FIG. 5 is a graph showing the variation of the carbon-nitrogen ratio between two groups.
FIG. 6 is a graph showing the change of germination index in two groups.
FIG. 7 is a graph showing the change of water content of two groups of mixed bacteria in the composting process.
FIG. 8 is a graph showing pH changes of two groups of mixed bacteria in the composting process.
FIG. 9 is a C/N change diagram of two groups of mixed bacteria in the composting process.
FIG. 10 is a diagram showing the germination index change of two groups of mixed bacteria in the composting process.
Detailed Description
First, preparation before experiment
1. Preparing fungus chaff, a feces sample and different strains; all strains can be purchased from the market.
B, bacillus subtilis: the number of effective viable bacteria of Jiangsu Lvke biotechnology limited company is 200 hundred million/g;
b, bacillus licheniformis: the number of effective viable bacteria of Jiangsu Lvke biotechnology limited company is 200 hundred million/g;
and (3) trichoderma viride: the number of effective live bacteria of Shandong and Zhongkang Yuan biological technology company Limited is 100 hundred million/g;
aspergillus niger: the number of effective live bacteria of Shandong and Zhongkang Yuan biological technology company Limited is 100 hundred million/g;
yeast: jiangsu green biotechnology limited company, 100 hundred million effective viable bacteria/g.
2. Preparing an instrument: mercury thermometer (1m), pH meter, sample crusher, conductivity meter, refrigerator at-4 deg.C, liquid nitrogen tank, drying oven at constant temperature of 105 deg.C, electronic balance and beaker.
3. Before the experiment is started, the water content, the pH value, the organic carbon, the total nitrogen, the carbon-nitrogen ratio, the ammonium nitrogen and the nitrate nitrogen in the sheep manure and the mushroom bran in the initial state are respectively measured.
Second, Experimental methods
1. Design of experiments
Influence of different microbial inoculum on sheep manure harmless treatment efficiency
The sheep manure is used as a main material, the mushroom dregs (namely mushroom bran) are used as an auxiliary material, the uniformly mixed materials are transported to a fermentation area to form a strip-stack-type stack body with two stacks of length, width and height being 1.5m multiplied by 1.5m, then adding EM microbial inoculum (purchased from Baiyibao biotechnology limited) purchased in the market, a compound microbial inoculum group 1 (bacillus subtilis: bacillus licheniformis: trichoderma viride: aspergillus niger: saccharomycetes) (in a mass ratio of 0.5:0.5:1:1:2) (all the microbial inocula are sold in the market) and a compound microbial inoculum group 2 (bacillus subtilis: bacillus licheniformis: trichoderma viride: aspergillus niger: saccharomycetes) (in a mass ratio of 1:1:1:2) (all the microbial inoculas are sold in the market) for composting, periodically measuring the temperature, the pH, the ammonium nitrogen and the nitrate nitrogen, finally judging the fertility through a germination index, the addition amount of the EM microbial inoculum, the complex microbial inoculum group 1 and the complex microbial inoculum group 2 in the compost is 4 per mill (w/w).
The test compares the composting effects of EM microbial inoculum purchased in the market and mixed microbial inoculum of different proportions, so as to judge the more appropriate proportion of the mixed microbial inoculum. The mixed bacteria agents with different proportions are a compound bacteria agent group 1 and a compound bacteria agent group 2.
2. Index and method for measuring the same
2.1 measurement indexes: temperature, pH, nitrogen, C/N, ammonium nitrogen, harmful heavy metals, germination index, and water content.
2.2 sampling method: the stack is divided into three parts according to the length, samples at each height are collected in an equal amount, then the samples are mixed evenly, and the samples are sampled for three times by a quartering method so as to be representative. One part was stored in a 4 ℃ freezer, one part was stored in a-80 ℃ freezer, and the other part was air-dried naturally for physical and chemical analysis.
2.3 determination method:
2.3.1 temperature:
the temperature of 5 points at the same height in the middle of the stack was randomly measured using mercury thermometers at 9:00 and 15:00 a day, and the average temperature was taken as the actual temperature of the stack.
2.3.2 pH and conductivity (EC):
taking 10g (dry mass) of a fresh compost sample, putting the fresh compost sample into a 200mL wide-mouth bottle, adding deionized water according to the mass ratio of 1:10, shaking the mixture on a shaking table for 2 hours, standing the mixture for 30min, and respectively measuring the pH value of the mixture by using a pH meter and a conductivity meter.
2.3.3 determination of seed Germination Index (GI):
mixing the decomposed compost fresh sample with distilled water according to the mass ratio of 1:10, fully oscillating, and leaching in a 30 ℃ thermostat for 24 hours. Sucking 7ml of filtrate, adding the filtrate into 9cm culture dishes paved with 2 pieces of 7cm qualitative filter paper, sowing 20 full pakchoi seeds in each culture dish, culturing in a constant-temperature incubator at 30 ℃ for 48h, measuring the germination rate and the root length of the seeds, and calculating the germination index GI (%). Triplicate for each treatment, distilled water was used as a control. The seed germination rate calculation formula is as follows (Guo et al.2012): GI (%) × (treatment average germination rate × treatment average root length)/(control average germination rate × control average root length) × 100.
2.3.4 Water content:
and (3) washing the aluminum box, drying the aluminum box in a constant-temperature drying box at the temperature of 100-105 ℃, and weighing and recording the mass A1. Weighing 5g (accurate to 0.001) of a fresh compost sample, putting the fresh compost sample into a drying aluminum box, transferring the fresh compost sample into a 105 ℃ constant-temperature drying box, drying the fresh compost sample for 24 hours, putting the fresh compost sample into a dryer for 30min, and taking out the fresh compost sample to be weighed and recorded with the mass A2.
The moisture content of compost sample is (A2-A1)/5 × 100%
Third, experimental results
The composting effects of the mixed bacteria agents with different proportions are compared, so that the more appropriate proportion of the mixed bacteria agents is judged. The mixed bacteria agents with different proportions are a compound bacteria agent group 1 and a compound bacteria agent group 2. From the graph 7, the two groups of water content have very obvious downward trends from the beginning to the end of composting, when the water content reaches about 30%, the fertilizer reaches primary maturity, the compound bacteria group 1 reaches primary maturity about day 37, and the compound bacteria group 2 reaches primary maturity about day 50.
Research shows that the pH value is between 8.0 and 9.0, which indicates that the compost is completely decomposed and meets the national standard. As can be seen from FIG. 8, the complex microbial inoculum group 1 is maintained between 8.0 and 9.0 during the composting period and meets the standard, and the pH of the complex microbial inoculum group 2 is slightly higher between 9.1 and 9.3 in the early and middle stages, but is reduced to below 9.0 in about 50 days.
The C/N plays an important role in composting, and the proper C/N can increase the relative abundance of bacillus and plays an important role in mesophilic and thermophilic stages. The abundance of bacillus is related to the activity of cellulose and beta-glycosidase, and the degradation and humification of the fiber can be improved by improving the activity of the cellulose and the beta-glycosidase. A study shows that the fertilizer is decomposed when the C/N is close to 15, and as can be seen from figure 9, the C/N in both groups reaches below 15 in the composting period, which indicates that the fertilizer is basically decomposed. However, it is considered that the C/N is not accurate to judge whether the fertilizer is decomposed, so more people can judge the decomposition of the fertilizer together with the germination index at the present stage.
The germination index is an important index for evaluating the compost maturity, and a large number of researches show that due to different compost materials, the C/N is used for evaluating whether the compost is decomposed inaccurately, and the judgment of whether the compost is decomposed by the germination condition and toxicity of plant seeds through compost leaching liquor is widely accepted. Generally, the germination index of more than 80% indicates that the fertilizer is completely decomposed, and more than 110% indicates that the fertilizer is completely decomposed, and as shown in fig. 10, the compound microbial inoculum group 1 reaches 110% in about 35 days to indicate that the fertilizer is completely decomposed, while the germination index of the compound microbial inoculum group 2 is maintained between 60% and 80% for most of the time, which indicates that the fertilizer is basically decomposed.
The experimental results show that: in the composting process, the composting of the composite microbial inoculum group 1 (bacillus subtilis: bacillus licheniformis: trichoderma viride: aspergillus niger: saccharomycetes) can promote the fertilizer to be thoroughly decomposed faster and better than the composting of the composite microbial inoculum group 2 (bacillus subtilis: bacillus licheniformis: trichoderma viride: aspergillus niger: saccharomycetes: 1:1:1: 2); the compounding effect of the compound microbial inoculum is obviously influenced by different ratios of strains in the compound microbial inoculum.
The compost composite microbial inoculum group 1 provided by the invention is prepared from bacillus subtilis, bacillus licheniformis, trichoderma viride, aspergillus niger and saccharomycetes in a microbial inoculum ratio of 0.5:0.5:1:1:2, and the moulds (aspergillus niger) and saccharomycetes with higher proportions in the composition have obvious effects on reducing the loss of total nitrogen and organic nitrogen and promoting maturity. Moreover, because the optimal temperatures of different strains and the temperature intervals capable of playing roles are different, for example, the high-temperature resistance characteristic of the bacillus subtilis plays an important role in compost fermentation but only plays a role in the high-temperature period of compost, and the function is limited, the bacillus subtilis needs to be cooperated with the mixture ratio of normal-temperature bacteria, mold, actinomycetes and the like to form a fermented fertilizer with higher efficiency of the composite microbial inoculum, so that the waste is recycled.
The composite microbial inoculum group 1 screened by the method is compared with commercial microbial inocula for composting, sheep manure is also selected for composting, and mushroom bran is selected as a bottom material. The initial values of the various materials are detected before composting.
TABLE 1-1 initial values of sheep manure and mushroom bran detection data table
Mixing mushroom bran and sheep manure, adjusting C/N to 20-25, composting, using the sheep manure as a main material, using mushroom dregs as an auxiliary material, transferring the uniformly mixed material to a fermentation area to form two stacks of strip-shaped stacks with the length multiplied by the width multiplied by the height multiplied by 1.5m, then respectively adding a microbial inoculum purchased in the market and a composite microbial inoculum (bacillus subtilis: trichoderma viride: aspergillus niger: saccharomycetes: 0.5:1:1:2) screened according to dominant strains for composting, periodically sampling and detecting the temperature, pH, ammonium nitrogen and nitrate nitrogen, and finally judging the fertility of the compost through a germination index. After about 60 days of composting and seven sampling and data detection, the results shown in figure 1 and figure 2 are finally obtained.
As can be seen from FIGS. 1 and 2, the early temperature of the mixed bacterium group rises faster than that of the commercial bacterium group, and the time for maintaining the high temperature is also higher than that of the commercial bacterium group, the rising speed of the early temperature can determine the decomposition speed of the fertilizer to a certain extent, and if the rising speed of the early temperature is fast, the time for reaching the decomposition is also shortened. According to the requirements of compost hygiene, bacteria and ova can be completely killed when the maximum compost temperature is more than 50-55 ℃ for 5-7 days, and both groups reach the harmless standard and are thoroughly decomposed.
When the moisture content reaches about 30%, the fertilizer reaches primary maturity, and as can be seen from fig. 3, the mixed bacteria group reaches primary maturity around day 37; the commercial bacterium group reaches primary maturity about day 50, and the water content of the mixed bacterium group is reduced faster in the early and middle stages because the temperature rise and the high temperature maintaining time of the mixed bacterium group are higher than those of the commercial bacterium group.
It is believed that a pH in the range of 8.0 to 9.0 meets the composting criteria, and both groups meet the criteria during composting (as shown in FIG. 4), and that in the early stages of composting, the moisture content is high, and the compost is locally anaerobic, and tends to produce small organic acids, which can reduce the pH.
The carbon-nitrogen ratio is a relatively important and common index for evaluating the compost maturity. About twenty-five carbon sources and one nitrogen source are available for microorganisms to synthesize one cell mass, so that composting is more favored by an initial C/N ratio of about 25 for the composting raw materials. It is generally considered that compost is decomposed when the C/N ratio is lower than 20, but the C/N ratio is different due to different bottom materials, so that some other indexes need to be combined for judgment. As can be seen from FIG. 5, the C/N of both compost is less than 20 at about day 37, indicating that the compost has reached maturity. The C/N of the compost using the mixed bacteria has a very obvious reduction in the early stage, and the C/N is obviously lower than that of the commercial bacteria at the beginning of the middle stage, which indicates that the nitrogen-preserving effect of the compost using the mixed bacteria is higher than that of the commercial bacteria.
As can be seen from fig. 6, the germination index during the composting process shows a tendency of first decreasing and then increasing, which may be due to high moisture content of the compost starting material, which may cause a local anaerobic state to generate small molecular organic acids, resulting in a decrease in the germination rate of the seeds. In the experiment, the germination indexes of the two groups reach 110% in about 50 days, and the two groups can be considered to be thoroughly decomposed in about 50 days. Wherein the feces, mushroom bran and mixed mushroom group reaches 110 percent firstly, and about 35 days later. From the above figure, it is obvious that the germination index of the fertilizer added with the mixed microbial inoculum is obviously higher than that of the fertilizer added with the commercial microbial inoculum. Therefore, the corrosion-promoting capability of the fertilizer added with the mixed microbial inoculum is stronger than that of the commercial microbial inoculum.
Claims (6)
1. The compost composite microbial inoculum is characterized by comprising bacillus subtilis, bacillus licheniformis, trichoderma viride, aspergillus niger and saccharomycetes, wherein the bacillus subtilis comprises the following components in percentage by mass: b, bacillus licheniformis: and (3) trichoderma viride: aspergillus niger: 0.2-0.8% of yeast: 0.2-0.8: 0.8-1.2: 0.8-1.2: 1.8 to 2.2.
2. A compost composite microbial inoculum according to claim 1, characterized in that the ratio by mass of bacillus subtilis: b, bacillus licheniformis: and (3) trichoderma viride: aspergillus niger: yeast 0.5:0.5:1:1: 2.
3. A compost composite bacterial agent as claimed in claim 1 or 2, wherein the effective viable count of said bacillus subtilis is 200 hundred million/g; the number of effective live bacteria of the bacillus licheniformis is 200 hundred million/g, the number of effective live bacteria of the trichoderma viride is 100 hundred million/g, and the number of effective live bacteria of the aspergillus niger is 100 hundred million/g; the effective viable count of the yeast is 100 hundred million/g.
4. Use of the complex microbial agent of claim 1, 2 or 3 in composting.
5. The use of claim 4, wherein the complex microbial inoculum is added into the compost in a mass ratio of 2-6%.
6. The use of claim 5, wherein the complex microbial inoculum is added into the compost in an amount of 4% by mass.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113135777A (en) * | 2021-04-22 | 2021-07-20 | 广西壮族自治区农业科学院 | On-site harmless treatment method for fruit and vegetable wastes |
| CN115011519A (en) * | 2022-06-13 | 2022-09-06 | 黄冈师范学院 | Composite microbial inoculum for degrading mushroom bran, mushroom bran fertilizer and preparation method thereof |
| CN115386506A (en) * | 2021-04-16 | 2022-11-25 | 中国科学院亚热带农业生态研究所 | Microbial compound microbial inoculum for promoting rapid composting of livestock and poultry manure and straw and method for producing bio-organic fertilizer by using microbial compound microbial inoculum |
| CN116987641A (en) * | 2023-08-07 | 2023-11-03 | 湖南省微生物研究院 | Corrosion-promoting nitrogen-preserving composting microbial inoculum and preparation method and application thereof |
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