Liquid culture medium for cultivating high-efficiency composite bacterial strain for composting and application thereof

Abstract

The application provides a kind of liquid culture medium for cultivating high-efficiency composite microbial system for compost and application thereof, and belongs to the technical field of microbial culture;The liquid culture medium includes microbial culture base medium and the following components: organic solid waste, self-inducer-2 (AI-2), N-acyl homoserine lactone (AHLs), vitamin B1, biotin and L-cysteine;The liquid culture medium takes organic solid waste, the target degradable substance, as carbon source;AI-2 and AHLs are used as signal molecules;Vitamin B1, biotin and L-cysteine are used as growth factors;Through the synergistic effect of characteristic carbon source, signal molecule and growth factor in microbial culture medium, the cultivation of high-efficiency composite microbial system for compost containing unculturable microorganisms in inoculum is realized, the number of available functional strains in organic solid waste composting system is greatly enriched and improved, and the interaction and degradation efficiency are strengthened.

Description

Technical Field

[0001] This invention belongs to the field of microbial culture technology, and in particular relates to a liquid culture medium for cultivating a highly efficient composite microbial system for composting and its application. Background Technology

[0002] Composting can transform unstable organic matter in solid waste into stable humus, reduce plant toxicity, promote nutrient recycling, and achieve a win-win situation for both the economy and the environment.

[0003] Currently, organic solid waste composting suffers from problems such as long composting cycles, low humification levels, and severe greenhouse gas and odor pollution. Adding relevant microbial agents during the organic solid waste composting process holds promise for solving these problems. Therefore, many researchers have employed strategies such as adding exogenous single strains or compound microbial agents to improve the quality and efficiency of organic solid waste composting.

[0004] However, whether using a single strain or a compound microbial agent, microorganisms need to be enriched, purified, and cultured before utilization. This method can only screen out culturable microorganisms; unculturable microorganisms and those requiring symbiotic relationships with other microorganisms are eliminated during the purification and culture stages. Furthermore, the carbon source in traditional culture media (such as LB broth and beef extract peptone) is mainly glucose or starch, meaning that microorganisms screened using these media often cannot effectively utilize the carbon source in the target composting system. Therefore, microorganisms screened using traditional media often have limited functions and categories, with many functional microorganisms lost and unable to be effectively utilized. Summary of the Invention

[0005] The purpose of this invention is to provide a liquid culture medium for cultivating a highly efficient compound microbial system for composting and its application, which can be used to cultivate a highly efficient compound microbial system for composting organic solid waste.

[0006] This invention provides a liquid culture medium for a highly efficient composite microbial system used in organic solid waste composting, comprising a basic culture medium for microbial culture and components at the following concentrations: organic solid waste 5-10 g / L, self-inducing agent-210-50 μg / L, N-acylhomoserine lactone 1-5 μg / L, vitamin B1 0.3-1 mg / L, biotin 0.1-0.4 mg / L, and L-cysteine ​​20-40 mg / L; the pH of the liquid culture medium is 6.5-7.5.

[0007] Preferably, the liquid culture medium further includes byproducts of organic solid waste composting; the byproducts of composting include compost leachate.

[0008] Preferably, the liquid culture medium comprises the following components at the following concentrations: organic solid waste 7.5–10 g / L, AI-2 30–50 μg / L, N-acylhomoserine lactone 3–5 μg / L, vitamin B1 0.5–1 mg / L, biotin 0.2–0.4 mg / L, and L-cysteine ​​20–40 mg / L; the concentration of compost leachate in the liquid culture medium is 50–100 mL / L.

[0009] Preferably, the organic solid waste includes one or more of the following: kitchen waste, straw, landscaping waste, and livestock and poultry manure.

[0010] The present invention also provides the application of the liquid culture medium described above in at least one of the following: screening, enrichment, cultivation and utilization of highly efficient compound microbial strains for composting.

[0011] Preferably, the high-efficiency composite microbial system for composting includes composite functional microorganisms; the composite functional microorganisms include one or more species at the level of the genera Bacillus, Parageobacillus, Sulfurovum, Gordonia, Georgenia, Pseudomonas, and Thiobacillus.

[0012] Preferably, the composite functional microorganisms include microbial communities related to organic matter degradation and / or microbial communities related to greenhouse and odorous gas regulation; the microbial communities related to organic matter degradation and transformation include one or more of the following genera: Bacillus, Pseudomonas, and Parageobacillus; the microbial communities related to greenhouse and odorous gas regulation include one or more of the following genera: Sulfurovum, Gordonia, and Thiobacillus.

[0013] The present invention also provides a highly efficient composite microbial system for composting, which is obtained by culturing the liquid culture medium described in the above scheme.

[0014] The present invention also provides a method for screening, enriching and / or culturing a highly efficient compound microbial system for composting, comprising: inoculating composting products into the liquid culture medium described in the above scheme for cultivation to obtain a culture containing a highly efficient compound microbial system for composting; wherein the composting raw material of the composting products and the target degradation products of the highly efficient compound microbial system for composting are of the same type.

[0015] The present invention also provides the application of the high-efficiency composite microbial system for composting described in the above scheme or the culture obtained by the method in the composting of organic solid waste.

[0016] This invention provides a liquid culture medium for cultivating a highly efficient composite microbial system for composting, comprising a basic culture medium for microbial culture and components at the following concentrations: organic solid waste 5-10 g / L, autoinducer-2 (AI-2) 10-50 μg / L, N-acylhomoserine lactones (AHLs) 1-5 μg / L, vitamin B1 0.3-1 mg / L, biotin 0.1-0.4 mg / L, and L-cysteine ​​20-40 mg / L; the pH of the liquid culture medium is 6.5-7.5. The liquid culture medium of this invention uses organic solid waste as a carbon source; it uses AI-2 and AHLs as signaling molecules to regulate the functional microbial community in the anaerobic digestion system, enhance microbial interactions, and promote the secretion and activity of key enzymes; it uses vitamin B1, biotin, and L-cysteine ​​as growth factors to form a synergistic mechanism through energy metabolism support, membrane function regulation, and redox environment optimization, thereby enabling the microbial community to degrade complex organic matter and achieving efficient enrichment of functional microbial communities through metabolic selection pressure; the liquid culture medium of this invention utilizes unique carbon... The synergistic effect of source, signaling molecules and growth factors enables the cultivation of highly efficient complex microbial systems, including unculturable microorganisms, in composting products (such as functional microorganisms at the level of Bacillus, Parageobacillus, Sulfurovum, Gordonia, Georgenia and Pseudomonas), which can greatly improve and enrich the available functional strains in organic solid waste composting. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 To show the growth rate of microorganisms under different culture media;

[0019] Figure 2 The Shannon index is used for microbial diversity analysis.

[0020] Figure 3 The Chao1 index was used for microbial diversity analysis.

[0021] Figure 4 The composition of microbial communities under different microbial culture media;

[0022] Figure 5 The effect of microorganisms screened for different microbial culture media on the seed germination index of composting systems;

[0023] Figure 6 The effect of microorganisms screened for different microbial culture media on the degree of polymerization (DP) of compost products;

[0024] Figure 7 The impact of microorganisms screened for different microbial culture media on greenhouse gas emissions during composting; where a represents cumulative N2O emissions and b represents cumulative CH4 emissions;

[0025] Figure 8 The effects of microorganisms screened for different microbial culture media on odor emissions during composting; where a represents the cumulative emissions of NH3, b represents the cumulative emissions of Me2S, c represents the cumulative emissions of Me2SS, and d represents the cumulative emissions of H2S. Detailed Implementation

[0026] This invention provides a liquid culture medium for cultivating a highly efficient composite microbial system for composting, comprising a basic culture medium for microbial culture and components at the following concentrations: organic solid waste 5-10 g / L, self-inducing agent-210-50 μg / L, N-acylhomoserine lactone 1-5 μg / L, vitamin B1 0.3-1 mg / L, biotin 0.1-0.4 mg / L, and L-cysteine ​​20-40 mg / L; the pH of the liquid culture medium is 6.5-7.5.

[0027] In one implementation, the organic solid waste is the target degradation product for composting.

[0028] The liquid culture medium of this invention is used to enrich a highly efficient composting functional microbial community, including unculturable microorganisms. The prepared highly efficient composting functional microbial community is used to shorten the composting process of organic solid waste, promote composting and humification, and reduce the emission of greenhouse gases and odorous gases during the composting process of organic solid waste.

[0029] Traditional microbial culture media struggle to cultivate highly efficient functional microorganisms because: 1) the carbon source in traditional media is inconsistent with the target degradation product, preventing the selected microbial communities from adapting well to the composting system. This invention directly uses the target degradation product as a specific carbon source, screening for strains adapted to this carbon source at the source; 2) traditional media neglect the synergistic metabolic relationships between microorganisms. This invention uses AI-2 and AHLs as signaling molecules to regulate functional microbial communities in the anaerobic digestion system, enhancing microbial interactions and promoting the secretion and activity of key enzymes. Using vitamin B1, biotin, and L-cysteine ​​as growth factors, a synergistic mechanism is formed through energy metabolism support, membrane function regulation, and redox environment optimization, enabling the microbial communities to degrade complex organic matter. Furthermore, metabolic selection pressure achieves efficient enrichment of functional microbial communities. Additionally, the traditional application model of culture media is "enrichment-separation-purification-expansion," which only yields a single culturable microbial strain. This invention uses composted products as inoculum, selects highly efficient functional microbial communities from them using a unique carbon source, and enhances their interaction and metabolic capabilities with signaling molecules and growth factors, thereby enabling in-situ culture of highly efficient complex microbial systems, including unculturable microorganisms.

[0030] The liquid culture medium of the present invention is a special culture medium for composting microbial agents based on highly efficient composite microbial enrichment and functional synergy.

[0031] In one embodiment, the liquid culture medium further includes byproducts of composting organic solid waste; the byproducts of composting include compost leachate.

[0032] In this invention, the compost leachate contains lignin and its degradation products (e.g., gallic acid, protocatechuic acid, etc.), polyphenolic compounds (e.g., caffeic acid, p-hydroxybenzoic acid, chlorogenic acid, etc.), amino acids and reducing sugars, polysaccharides, organic acids, aromatic groups (e.g., phenylalanine and its derivatives), and protein decomposition products (amino acids and other small molecule organic matter). The liquid culture medium of this invention uses compost byproducts as functional enhancers, providing trace elements, humic acid precursors, and cell secretions. The humic acid precursors and trace elements activate oxidases to promote humic aggregation and activate the expression of key microbial functional genes. Simultaneously, its organic acids inhibit sulfate-reducing bacteria activity, reducing sulfide formation, and promote sulfur oxidation by providing electron acceptors such as nitrates. The key microbial functional genes include humification-related genes, nitrogen metabolism genes, and carbon metabolism and organic matter degradation genes.

[0033] In one embodiment, the liquid culture medium comprises the following components at the following concentrations: organic solid waste 7.5–10 g / L, AI-2 30–50 μg / L, AHLs 3–5 μg / L, vitamin B1 0.5–1 mg / L, biotin 0.2–0.4 mg / L, and L-cysteine ​​20–40 mg / L; the concentration of compost leachate in the liquid culture medium is 50–100 mL / L.

[0034] As one embodiment, the basic culture medium for microbial culture, based on 1L of liquid culture medium, comprises the following components: 0.5 g of (NH4)2SO4, 0.25 g of K2HPO4, 0.125 g of MgSO4, 0.125 g of NaCl, 0.0025 g of MnSO4·4H2O, and 60 mg of FeSO4·7H2O.

[0035] In one embodiment of the present invention, the liquid culture medium consists of a basic culture medium for microbial culture and components at the following concentrations: 7.5 g / L organic solid waste, 250 μg / L self-inducing agent, 3 μg / L N-acylhomoserine lactone, 1 mg / L vitamin B1, 0.4 mg / L biotin, 20 mg / L L-cysteine, and 100 mL / L compost leachate.

[0036] In one embodiment of the present invention, the liquid culture medium consists of a basic culture medium for microbial culture and components at the following concentrations: 7.5 g / L organic solid waste, 250 μg / L self-inducing agent, 3 μg / L N-acylhomoserine lactone, 1 mg / L vitamin B1, 0.4 mg / L biotin, and 20 mg / L L-cysteine.

[0037] In another embodiment of the invention, the liquid culture medium consists of a basic culture medium for microbial culture and components at the following concentrations: 5 g / L organic solid waste, 10 μg / L self-inducing agent-2, 1 μg / L N-acylhomoserine lactone, 0.3 mg / L vitamin B1, 0.1 mg / L biotin, 10 mg / L L-cysteine, and 50 mL / L compost leachate.

[0038] In another embodiment of the invention, the liquid culture medium consists of a basic culture medium for microbial culture and components at the following concentrations: 7.5 g / L organic solid waste, 230 μg / L self-inducing agent, 3 μg / L N-acylhomoserine lactone, 0.5 mg / L vitamin B1, 0.2 mg / L biotin, 20 mg / L L-cysteine, and 75 mL / L compost leachate.

[0039] In another embodiment of the invention, the liquid culture medium consists of a basic culture medium for microbial culture and components at the following concentrations: 10 g / L organic solid waste, 250 μg / L self-inducing agent, 5 μg / L N-acylhomoserine lactone, 1 mg / L vitamin B1, 0.4 mg / L biotin, 40 mg / L L-cysteine, and 100 mL / L compost leachate.

[0040] As one implementation method, the organic solid waste includes one or more of kitchen waste, straw, garden waste and livestock manure, and is further defined as kitchen waste.

[0041] The present invention also provides the application of the liquid culture medium described above in at least one of the following: screening, enrichment, cultivation and utilization of highly efficient compound microbial strains for composting.

[0042] In one embodiment, the highly efficient composite microbial system for composting includes culturable and / or non-culturable composite functional microorganisms; the composite functional microorganisms include one or more species at the level of Bacillus, Parageobacillus, Sulfurovum, Gordonia, Georgenia, Pseudomonas, and Thiobacillus; functionally, the composite functional microorganisms include those related to organic matter... The microbial communities related to organic matter degradation and / or related to greenhouse and odor control; the microbial communities related to organic matter degradation and transformation include one or more of the genera Bacillus, Pseudomonas, and Parageobacillus; the microbial communities related to greenhouse and odor control include one or more of the genera Sulfurovum, Gordonia, and Thiobacillus.

[0043] In one embodiment, the high-efficiency composite microbial system for composting includes a high-efficiency composite microbial system in the composting products.

[0044] The liquid culture medium of this invention achieves the cultivation of functional microorganisms (such as functional microorganisms of the genera Bacillus, Parageobacillus, Sulfurovum, Gordonia, Georgeia, and Pseudomonas) in composting products, including unculturable microorganisms, through the synergistic effect of organic solid waste, signaling molecules, growth factors, and functional enhancers. This greatly enhances and enriches the available functional strains in organic solid waste composting. The liquid culture medium of this invention can increase the abundance of functional microorganisms (composting-enhancing microorganisms) in microbial agents, such as Bacillus, Parageobacillus, Sulfurovum, Gordonia, and Georgeia, by 16.67%–66.67%, 40.00%–73.33%, 12.50%–50.00%, 58.33%–66.67%, and 20.00%–70.00%, respectively. This increase in the relative abundance of these functional microorganisms can improve the composting efficiency of potential organic solid waste. The liquid culture medium of this invention can effectively improve the community diversity and richness of highly efficient composite microbial systems for composting.

[0045] In one embodiment, the use of a highly efficient composite microbial system for composting includes promoting compost humification and / or reducing odor and greenhouse gas emissions; the odorous gases include those containing sulfur and nitrogen; and the greenhouse gases include those containing nitrogen and carbon.

[0046] The microbial community cultured in the liquid culture medium of the present invention can effectively shorten the composting cycle, increase the degree of compost humification, and reduce the emission of odors and greenhouse gases.

[0047] The present invention also provides a highly efficient composite microbial system for composting, which is obtained by culturing the liquid culture medium described in the above scheme.

[0048] In one embodiment, the high-efficiency composite microbial system for composting uses the composted product of organic solid waste as the microbial inoculum source and is cultured in the liquid culture medium; the composting raw material of the composted product and the target degradation products of the high-efficiency composite microbial system for composting are of the same type.

[0049] This invention also provides a method for screening, enriching, and / or cultivating highly efficient compound microbial strains for composting, comprising:

[0050] The composting products are inoculated into the liquid culture medium described in the above scheme and cultured to obtain a culture containing a high-efficiency compound microbial system for composting; the composting raw materials of the composting products and the target degradation products of the high-efficiency compound microbial system for composting are of the same type.

[0051] The method of this invention uses composting products as microbial inoculum, selects highly efficient functional microbial communities from them using a special carbon source, and enhances their interaction and metabolic capabilities with signaling molecules and growth factors. It can screen, enrich and cultivate key functional microbial communities that promote the composting of organic solid waste in composting products, and can realize in-situ cultivation of highly efficient complex microbial systems including unculturable microorganisms.

[0052] In one implementation, the target degradation product of the high-efficiency composite microbial system for composting is kitchen waste, and the composting raw material of the composting maturation product is also kitchen waste; the composting raw material of the composting maturation product and the target degradation product of the high-efficiency composite microbial system for composting can be of the same type, and are not limited to being from the same source.

[0053] In this invention, the ratio of the mass of the composting product to the volume of the liquid culture medium is 2g:100mL. The method of this invention can screen, enrich, and cultivate highly efficient composite microbial strains for composting.

[0054] In one embodiment, the culture temperature is 30°C; the culture time is 48 hours; the culture is a shaking culture; and the shaking rotation speed is 180 rpm.

[0055] The method of this invention produces a high-efficiency compound microbial strain for composting with a fast growth rate, which is beneficial to the recovery and growth of the high-efficiency compound microbial strain for composting.

[0056] The present invention also provides microbial agents obtained by liquid culture medium culture as described above, or cultures obtained by the method described above.

[0057] The present invention also provides the application of the high-efficiency composite microbial system for composting described in the above scheme or the culture obtained by the method in the composting of organic solid waste.

[0058] The microbial agents and cultures of the present invention can improve the composting efficiency of organic solid waste and the quality of composting products.

[0059] The present invention also provides a method for composting organic solid waste, comprising the following steps:

[0060] The organic solid waste is inoculated with the high-efficiency compound microbial system described in the above scheme or the culture obtained by the method described above, and composting is carried out.

[0061] To further illustrate the present invention, the following detailed description, in conjunction with the accompanying drawings and embodiments, provides a liquid culture medium for cultivating a highly efficient composite microbial system for composting and its application, but these descriptions should not be construed as limiting the scope of protection of the present invention.

[0062] Example 1: A liquid culture medium and its preparation for cultivating a highly efficient composite microbial strain for kitchen waste composting.

[0063] (1) Raw materials

[0064] It consists of basal culture medium, a unique carbon source, signaling molecules, growth factors, and functional enhancers. The specific composition and content are as follows:

[0065] ①Basic culture medium: (NH4)2SO4 0.5 g / L, K2HPO4 0.25 g / L, MgSO4 0.125 g / L, NaCl 0.125 g / L, MnSO4·4H2O 0.0025 g / L and FeSO4·7H2O 60 mg / L).

[0066] ② Unique carbon source: 7.5g / L of kitchen waste;

[0067] ③ Signaling molecules: AI-2 self-inducible molecules: 50 μg / L; N-acylhomoserine lactones (AHLs): 3 μg / L.

[0068] ④ Growth factors: Vitamin B1 1.0 mg / L, Biotin 0.4 mg / L, L-cysteine ​​20 mg / L.

[0069] ⑤ Functional enhancer: Leachate from kitchen waste composting: 100mL / L.

[0070] (2) Culture medium preparation

[0071] Prepare 1000 mL of liquid culture medium according to the above formula and adjust the pH to 6.5–7.5. Example 2: A liquid culture medium for cultivating a highly efficient composite microbial strain for kitchen waste composting and its preparation.

[0072] (1) Raw materials

[0073] It consists of basal culture medium, a unique carbon source, signaling molecules, growth factors, and functional enhancers. The specific composition and content are as follows:

[0074] ①Basal culture medium: Same as in Example 1.

[0075] ②Special carbon source: 5g / L of kitchen waste;

[0076] ③ Signaling molecules: AI-2 self-inducible molecules: 10 μg / L; N-acylhomoserine lactones (AHLs): 1 μg / L.

[0077] ④ Growth factors: Vitamin B1 0.3 mg / L, Biotin 0.1 mg / L, L-cysteine ​​10 mg / L.

[0078] ⑤ Functional enhancer: Leachate from kitchen waste composting: 50mL / L.

[0079] (2) Culture medium preparation

[0080] Prepare 1000 mL of liquid culture medium according to the above formula and adjust the pH to 6.5–7.5. Example 3: A liquid culture medium for cultivating a highly efficient composite microbial strain for kitchen waste composting and its preparation.

[0081] (1) Raw materials

[0082] It consists of basal culture medium, a unique carbon source, signaling molecules, growth factors, and functional enhancers. The specific composition and content are as follows:

[0083] ①Basal culture medium: Same as in Example 1.

[0084] ② Unique carbon source: 7.5g / L of kitchen waste;

[0085] ③ Signaling molecules: AI-2 self-inducible molecules: 30 μg / L; N-acyl homoserine lactones (AHLs): 3 μg / L.

[0086] ④ Growth factors: Vitamin B1 0.5 mg / L, Biotin 0.2 mg / L, L-cysteine ​​20 mg / L.

[0087] ⑤ Functional enhancer: Leachate from kitchen waste composting: 75 mL / L.

[0088] (2) Culture medium preparation

[0089] Prepare 1000 mL of liquid culture medium according to the above formula and adjust the pH to 6.5–7.5. Example 4: A liquid culture medium for cultivating a highly efficient composite microbial strain for kitchen waste composting and its preparation.

[0090] (1) Raw materials

[0091] It consists of basal culture medium, a unique carbon source, signaling molecules, growth factors, and functional enhancers. The specific composition and content are as follows:

[0092] ①Basal culture medium: Same as in Example 1.

[0093] ②Special carbon source: 10g / L of kitchen waste;

[0094] ③ Signaling molecules: AI-2 self-inducible molecules: 50 μg / L; N-acylhomoserine lactones (AHLs): 5 μg / L.

[0095] ④ Growth factors: Vitamin B1 1.0 mg / L, Biotin 0.4 mg / L, L-cysteine ​​40 mg / L.

[0096] ⑤ Functional enhancer: Leachate from kitchen waste composting: 100mL / L.

[0097] (2) Culture medium preparation

[0098] Prepare 1000 mL of liquid culture medium according to the above formula and adjust the pH to 6.5-7.5. Example 5: A liquid culture medium and its preparation for cultivating a highly efficient compound microbial strain for kitchen waste composting are the same as in Example 1, except that the functional enhancer is omitted.

[0099] Comparative Example 1: Traditional carbon source culture medium and its preparation

[0100] It consists of a basal culture medium and sodium succinate, with sodium succinate serving as the carbon source.

[0101] The configuration process is the same as in Example 1.

[0102] Comparative Example 2

[0103] Except for omitting the signal molecule, the rest is the same as in Example 1.

[0104] Comparative Example 3

[0105] Except for omitting the growth factor, the rest is the same as in Example 1.

[0106] Experimental Example 1: Microbial Cultivation of Kitchen Waste

[0107] Highly efficient complex microbial strains in the composting products of kitchen waste were screened, enriched, and cultured using the liquid culture media of Examples 1-5 and Comparative Examples 1-3, respectively. The composting products of kitchen waste were inoculated into the liquid culture media of Examples 1-5 and Comparative Examples 1-3 at an inoculation concentration of 20 g / L. The cultures were incubated at 30°C with shaking at 180 rpm for 48 h. After incubation, the content of humus, sulfides in the water, and microbial biomass (OD) were measured. 600 ).

[0108] The results are shown in Table 1.

[0109] Table 1. Effects of differences in microbial culture medium composition on organic waste humus, water sulfides, and biomass.

[0110] Comparative Example 1 <![CDATA[180.12±25.96 e ]]> <![CDATA[1.03±0.24 de ]]> <![CDATA[2.2±0.12 a ]]> Comparative Example 2 <![CDATA[200.47±13.5 e ]]> <![CDATA[0.85±0.22 e ]]> <![CDATA[1.80±0.12 cd ]]> Comparative Example 3 <![CDATA[218.42±11.8 de ]]> <![CDATA[0.92±0.20 e ]]> <![CDATA[1.91±0.21 abc ]]> Example 1 <![CDATA[295.78±24.5 a ]]> <![CDATA[1.70±0.13 a ]]> <![CDATA[1.95±0.31 ab ]]> Example 2 <![CDATA[255.71±21.5 b ]]> <![CDATA[1.60±0.28 ab ]]> <![CDATA[1.42±0.15 e ]]> Example 3 <![CDATA[248.90±18.9 bc ]]> <![CDATA[1.25±0.15 cd ]]> <![CDATA[1.63±0.10 e ]]> Example 4 <![CDATA[235.22±16.3 cd ]]> <![CDATA[1.45±0.24 c ]]> <![CDATA[1.71±0.17 de <!-- 7 -->]]> Example 5 <![CDATA[230.61±12.1 cd ]]> <![CDATA[1.64±0.31 a ]]> <![CDATA[1.87±0.12 bc ]]>

[0111] Note: ANOVA analysis and Tukey HSD post-hoc test were used, with a significance level of α = 0.05.

[0112] As can be seen from Table 1, although the OD of Example 1 was lower than that of Comparative Example 1, Example 1 showed a lower OD. 600 While the levels of carbon sources decreased, they significantly promoted humus formation and the retention of sulfides in the water. This demonstrates that while traditional carbon sources in microbial culture media can increase microbial biomass (OD), they also significantly promote humus formation and the retention of sulfides in the water. 600 However, due to the use of traditional carbon sources, the microorganisms that accumulate are likely to metabolize easily degradable organic matter, resulting in a poor ability to promote the composting of kitchen waste. Compared with Example 1, Comparative Examples 2 and 3 lacked signal molecules and growth factors in their microbial culture media. The absence of these components led to a decrease in the content of humic substances and water sulfides by 26.15–32.22% and 45.88–50.00%, respectively, indicating that these two components have a substantial effect on promoting the composting and deodorizing of organic solid waste. In contrast, Example 5 lacked a functional enhancer, resulting in a decrease in the content of humic substances and the retention of water sulfides, although not as high as in Example 1, but an increase of 5.58–28.03% and 59.22–92.94%, respectively, compared with Comparative Examples 1–3. The difference between Examples 2–4 lies in the difference in the content of each component of the microbial culture media. As can be seen from the table, the difference in the content of each component of the microbial culture media compared with Example 1 led to a certain degree of weakening in the content of humic substances and the retention of water sulfides. However, compared with Comparative Examples 1-3, Examples 2-4 increased the humic content and water sulfide content by 7.69-41.97% and 21.36-88.24%, respectively.

[0113] Comparative Example 5: Traditional Liquid Culture Medium A

[0114] Formula: per 1000 mL, it consists of the following components: NH4Cl 0.4 g, MgSO4·7H2O 0.05 g, K2HPO4 0.2 g, NaCl 0.12 g, FeSO4·7H2O 0.01 g, MnSO4·H2O 0.01 g and peptone 5 g.

[0115] Comparative Example 6: Traditional Liquid Culture Medium B

[0116] Formula: per 1000 mL, it consists of the following components: NH4Cl 5g, KCl 5g, MgSO4 2.5g, CaCl2 1.85g, NaCl 3g, NaHCO3 4.2g, peptone 30g / L and meat extract 20g / L.

[0117] Comparative Example 7: Traditional Liquid Culture Medium C

[0118] Formula: per 1000 mL, it consists of the following components: 1.0 g K2HPO4·3H2O, 0.20 g KH2PO4, 0.05 g CaCl2, 0.5 g MgCl2, 0.01 g FeCl2, 1.0 g (NH4)2SO4, 5.0 g NaCl and 1.0 g yeast extract.

[0119] Comparative Example 8: Traditional Liquid Culture Medium D

[0120] Formula: per 1000 mL, it consists of the following components: Na2HPO4 1.2 g, K2HPO4 1.8 g, MgSO4·7H2O 0.1 g, (NH4)2SO4 0.1 g, CaCl2 0.03 g, FeCl3 0.02 g, MnSO4 0.02 g and Na2S2O3 1.0 g.

[0121] Experimental Example 2

[0122] The liquid culture medium (patented culture medium) of Example 1 and the liquid culture media A, B, C, and D of Comparative Examples 5-8 were used as microbial culture media, respectively. 20 g / L of kitchen waste composting product was used as inoculum, and the natural microbial flora was enriched through incubation at 30°C for 48 hours (150 rpm) on a shaker. During the microbial culture process, such as... Figure 1 As shown, the microbial growth curve was determined. The results showed that the liquid culture medium in Example 1 had a faster microbial growth rate compared to other culture media, with an OD of [value missing] after 12 hours of culture. 600 The pH reached 2.38, and after 18 hours, the natural microbial community entered a plateau phase, while the growth rate of microorganisms cultured in other media was relatively slow. This indicates that the liquid culture medium in Example 1 was more conducive to microbial recovery and growth.

[0123] Experimental Example 3

[0124] Sequencing analysis was performed on the microbial communities enriched in the liquid culture medium (patented medium) of Example 1 and the liquid culture media A, B, C, and D of Comparative Examples 5-8. The Shannon index and Chao1 index revealed significant differences in community diversity and community richness among the different liquid culture media. Figure 2 and Figure 3 ).from Figure 2 and Figure 3It can be seen that the Shannon index and Chao1 index of the liquid culture medium treatment group in Example 1 were higher than those of other groups, indicating that the liquid culture medium in Example 1 can effectively improve the diversity and richness of the microbial community of the natural inoculant. This is because the liquid culture medium in Example 1 contains not only basic nutrients, but also trace elements, growth factors and signaling molecules, which allow some unculturable microorganisms to be cultured and enriched.

[0125] Test Example 4

[0126] Metagenomic sequencing technology was used to determine the microbial communities enriched in the liquid culture medium (patented culture medium) of Example 1 and the liquid culture media A, B, C, and D of Comparative Examples 5-8. Figure 4 The results showed that the microorganisms in the liquid culture medium treatment group of Example 1 significantly increased the abundance of key functional microorganisms at the genera level, including Bacillus, Parageobacillus, Gordonia, and Georgenia, compared to other treatment groups. This may be due to the addition of kitchen waste as a carbon source or the addition of substances such as signaling molecules and growth factors. Furthermore, the microorganisms in the liquid culture medium treatment group of Example 1 contained relatively high abundances of microbial communities related to organic matter transformation (Bacillus, Parageobacillus), odor control, and greenhouse gas emission reduction (Sulfurovum, Gordonia, Georgenia, Thiobacillus).

[0127] Experiment 5: Verification of the efficacy of microbial culture in different microbial culture media

[0128] To further verify the ability of the liquid culture medium (patented culture medium) of Example 1 to cultivate key functional microorganisms in mature compost, this experiment verified the efficacy of the microorganisms enriched and cultured in the liquid culture medium of Example 1 and the liquid culture media A, B, C, and D of Comparative Examples 5-8. Specifically, the microbial agents prepared from the liquid culture medium of Example 1 and the liquid culture media A, B, C, and D of Comparative Examples 5-8 were inoculated into kitchen waste at an inoculation rate of 10 mL / kg (v / w) and a composting experiment was conducted for 15 days. Figures 5-8 The results showed:

[0129] The microbial inoculant cultured in the liquid culture medium of Example 1 shortened the composting cycle by 13.33% compared to other treatments. Regarding seed germination index (… Figure 5In Example 1, the liquid culture medium group showed >70% filtration efficiency on day 13 of composting, and by day 15 of composting, the filtration efficiency of the liquid culture medium in Example 1 (84.22%) was > that of culture medium A (72.63%) > culture medium B (72.57%) > culture medium C (72.17%) > culture medium D (71.32%). Regarding the degree of polymerization (DP)... Figure 6 The ratios of the liquid culture medium in Example 1 (2.5g) > Culture medium B (2.4g) > Culture medium C (2.4g) > Culture medium A (2.01g) > Culture medium D (1.9g) showed that the liquid culture medium in Example 1 increased DP by 4.02%–30.15%. Furthermore, the liquid culture medium group in Example 1 reduced odorous gas emissions of NH3 by 11.62%–41.89%, Me2S by 9.27%–30.19%, Me2SS by 9.45%–38.21%, and H2S by 12.16%–40.09%. Figure 8 ); while greenhouse gas emissions of N2O and CH4 will be reduced by 10.05%–13.49% and 6.33%–9.53%, respectively. Figure 7 The results show that the highly efficient composite microbial strain cultured using the liquid culture medium of Example 1 significantly shortens the composting cycle, increases the degree of humification in compost, and reduces the emission of odors and greenhouse gases in the compost.

[0130] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A liquid culture medium for a highly efficient composite microbial system used in the composting of organic solid waste, characterized in that, The medium includes a basal culture medium for microbial culture and the following components at the following concentrations: organic solid waste 5-10 g / L, self-inducing agent-2 10-50 μg / L, N-acylhomoserine lactone 1-5 μg / L, vitamin B1 0.3-1 mg / L, biotin 0.1-0.4 mg / L, and L-cysteine ​​20-40 mg / L; the pH of the liquid culture medium is 6.5-7.

5. The liquid culture medium also includes byproducts of organic solid waste composting; The byproducts of the organic solid waste composting include leachate from the composting of kitchen waste; the organic solid waste is kitchen waste.

2. The liquid culture medium according to claim 1, characterized in that, The liquid culture medium comprises the following components at the following concentrations: organic solid waste 7.5~10 g / L, AI-2 30~50 μg / L, N-acylhomoserine lactone 3~5 μg / L, vitamin B1 0.5~1 mg / L, biotin 0.2~0.4 mg / L, and L-cysteine ​​20~40 mg / L; the concentration of the compost leachate from the kitchen waste in the liquid culture medium is 50~100 mL / L.

3. The use of the liquid culture medium according to claim 1 or 2 in at least one of the following: screening, enrichment, cultivation and utilization of highly efficient compound microbial strains for composting; The high-efficiency compound microbial system for composting includes compound functional microorganisms; the compound functional microorganisms include one or more species at the level of Bacillus, Parageobacillus, Sulfurovum, Gordonia, and Georgenia.

4. The application according to claim 3, characterized in that, The composite functional microorganisms include microbial communities related to organic matter degradation and / or microbial communities related to greenhouse and odorous gas regulation; the microbial communities related to organic matter degradation and transformation include one or more of the genus Bacillus and Parageobacillus; the microbial communities related to greenhouse and odorous gas regulation include one or more of the genus Sulfurovum and Gordonia.

5. A highly efficient composite microbial system for composting, characterized in that, Obtained by cultivation in the liquid culture medium according to claim 1 or 2; The high-efficiency compound microbial system for composting includes compound functional microorganisms; the compound functional microorganisms include species at the level of Bacillus, Parageobacillus, Sulfurovum, Gordonia, and Georgenia.

6. A method for screening, enriching, and / or cultivating highly efficient compound microbial strains for composting, characterized in that, include: The composted product is inoculated into the liquid culture medium described in claim 1 or 2 and cultured to obtain a culture containing a highly efficient composite microbial system for composting. The composting raw materials of the composting products and the target degradation products of the high-efficiency composite microbial system for composting are the same. The composting product is the composting product of kitchen waste.

7. The application of the high-efficiency composite microbial system for composting according to claim 5 or the culture obtained by the method according to claim 6 in the composting of organic solid waste, characterized in that, The organic solid waste is kitchen waste.

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