CN111394287A - Preparation method of degrading microbial inoculum for kitchen waste treatment - Google Patents
Preparation method of degrading microbial inoculum for kitchen waste treatment Download PDFInfo
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Abstract
The invention provides a preparation method of a degrading microbial inoculum for treating kitchen waste, wherein the degrading microbial inoculum comprises bacillus subtilis, bacillus lysinate, bacillus licheniformis, trichoderma viride, candida glabrata, yarrowia lipolytica and a biosurfactant; the method comprises the following steps: step A, activation culture, step B, domestication culture: performing domestication culture on the six bacteria respectively, specifically performing step-by-step domestication culture for gradually increasing the concentration of sodium chloride for culture on each microorganism; c, liquid fermentation and enlarged culture, D, dehydration and drying, and E, and mixing uniformly. The degrading microbial inoculum prepared by the method can rapidly decompose macromolecular organic matters such as protein, grease, starch polysaccharide, cellulose, hemicellulose and the like in the kitchen waste, so that the macromolecular organic matters are rapidly stabilized, the kitchen waste degrading efficiency is improved, the space utilization rate of a fermentation bin is further improved, and the quality of organic fertilizers which are degradation products of the kitchen waste is improved.
Description
Technical Field
The invention relates to the field of municipal waste treatment, in particular to a preparation method of a degrading microbial inoculum for kitchen waste treatment.
Background
With the rapid growth of economy and the increasing consumption level of residents in China, the consumption of people in catering is increased day by day, and the rapid growth of kitchen waste is brought. In 2018, the national kitchen waste production is about 1.08 hundred million tons, the daily average yield reaches 29.2 million tons/day, and the annual composite growth rate reaches more than 8%.
Kitchen waste is a general term for food waste and kitchen waste. The food waste refers to waste materials generated in the food using process of various restaurants, hotels, schools, enterprises, institutions and dining halls, waste leftovers, waste edible oil, waste food residues and the like. The kitchen waste refers to waste materials generated in the process of processing kitchen food and overdue waste food, such as vegetable leaves, fruit peels, animal viscera and the like generated in a household kitchen.
Hogwash oil illegally extracted by kitchen waste contains a large amount of dangerous carcinogenic substances, wherein aflatoxin with high toxicity is one of the strongest chemical carcinogenic substances found at present, and the toxicity of aflatoxin is 100 times that of arsenic trioxide. The food processed and produced by the hogwash oil contains a large amount of benzene components and a plurality of other carcinogenic substances which are harmful to human bodies, has great harm to the health of the human bodies, and can cause various cancers such as liver cancer, gastric cancer, kidney cancer, intestinal cancer, breast cancer, ovarian cancer and the like after being eaten for a long time.
The kitchen waste has the characteristics of high water content, high organic matter, high grease, high salinity, easy decay and easy deterioration, the water accounts for 70-90% of the wet weight of the kitchen waste, the content of organic dry matters reaches more than 90% of the dry weight of the kitchen waste, wherein the total content of crude protein, crude fat, crude fiber and total starch accounts for more than 70% of the organic dry matters, the grease content accounts for 3-5% of the wet weight of the kitchen waste, the salinity accounts for 0.2-1.0% of the wet weight of the kitchen waste, and the salinity mainly contains Na+And Cl-Also contains a small amount of Ca2+、Mg2+And K+And the like.
The kitchen waste integrated treatment equipment takes kitchen waste as a raw material and carries out rapid decomposition on the kitchen waste through comprehensive treatment technologies such as sorting, crushing, squeezing and dewatering, oil-water separation, high-temperature aerobic fermentation of microorganisms and the like. The organic garbage is reduced by 80-85% in the process of front-stage pretreatment, sewage is merged into an urban pipe network or is treated on site and then is discharged after reaching the standard, grease is recycled, 15-20% of solid emissions are fermented into organic fertilizer through a fermentation bin at high temperature in an aerobic manner, and the generated odor is absorbed and converted to be discharged after reaching the standard, so that the reduction, the harmlessness, the stabilization and the resource utilization of the kitchen waste treatment are realized.
Generally, the kitchen waste in the fermentation chamber of the kitchen waste integrated treatment equipment needs to stay for at least 7-15 days to reach a basic decomposition state. At present, due to the fact that a fermentation bin of kitchen waste integrated treatment equipment on the market is limited in space, kitchen waste stays for 1-5 days, the kitchen waste stays for a short time, the decomposition degree is low, and phenomena of seedling burning, root burning and the like are easily caused. At this time, the composite rotten microbial inoculum can be added into the kitchen waste according to a certain proportion, so that the fermentation efficiency is improved, the fermentation time is shortened, and the defect of insufficient space of a fermentation bin is overcome.
The kitchen waste has high grease content, the grease wraps materials, the kitchen waste is not easy to degrade, the viscosity is high, and the materials are dense and airtight. At present, the decomposing microbial inoculum for kitchen waste compost fermentation has low efficiency of degrading grease, and the decomposing microbial inoculum seriously affects the high-temperature aerobic fermentation process of the kitchen waste, prolongs the decomposing time, prolongs the material retention time, reduces the space utilization rate of a fermentation bin, has low decomposing degree of organic fertilizer, and seriously affects the product quality.
High salt content of the kitchen waste can generate salt stress, high-concentration salt can inhibit the growth and biological activity of microorganisms, and the microorganisms can lose the biological activity in severe cases. The research finds that the salt has strong respiratory inhibition effect on aerobic microorganisms. High salinity content influences the high-temperature aerobic fermentation process of kitchen waste, leads to fermentation efficiency to reduce, prolongs the time of becoming thoroughly decomposed, leads to the material dwell time extension, and fermentation storehouse space utilization reduces.
The Chinese patent application CN201910548200.3 provides a complex microbial inoculum and a method for treating kitchen waste by using the same, wherein the complex microbial inoculum comprises the following components: irpex lacteus, rumen microorganisms, bacillus subtilis, bacillus cereus, bacillus stearothermophilus, klebsiella oxytoca, bacillus licheniformis and aspergillus niger. By using the composite microbial inoculum, the treatment equipment and the treatment method, more than 90% of organic garbage in the kitchen garbage can be degraded, and the treatment efficiency is high. But the method is still not suitable for degrading the kitchen waste with high salt content.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a degrading microbial inoculum for treating kitchen waste and a preparation method thereof, which can improve the degrading efficiency of the kitchen waste, further improve the space utilization rate of a fermentation bin, quickly decompose grease, reduce the inhibiting effect of salt on the activity of the microbial inoculum, improve the degrading efficiency and the decomposition degree of the kitchen waste and improve the quality of organic fertilizer products.
Therefore, the invention firstly provides a preparation method of a degrading microbial inoculum for treating kitchen waste, wherein the degrading microbial inoculum comprises bacillus subtilis, bacillus lysinate, bacillus licheniformis, trichoderma viride, candida glabrata, yarrowia lipolytica and a biosurfactant; the preparation method comprises the following steps: step A, activation culture: firstly, respectively inoculating six bacteria of bacillus subtilis, bacillus lysinate, bacillus licheniformis, trichoderma viride, candida glabrata and yarrowia lipolytica into activation culture media of the six bacteria for activation culture; step B, domestication culture: then, carrying out domestication culture on the six bacteria respectively, wherein the domestication culture is a step-by-step domestication culture for gradually increasing the concentration of sodium chloride for culture on each microorganism; c, liquid fermentation and enlarged culture: respectively inoculating six bacteria obtained by domestication into a fermentation tank of the six bacteria for liquid fermentation and expanded culture; step D, dehydration and drying: dehydrating and drying various single bacteria obtained by amplification culture to prepare dormant microorganism dry powder; step E, mixing uniformly: and D, uniformly mixing the six dormant body microorganism dry powders prepared in the step D, and uniformly mixing the six dormant body microorganism dry powders with a biosurfactant to obtain the degrading microbial inoculum for treating the kitchen waste.
In a specific embodiment, in the step D, 20-30 parts of bacillus subtilis, 10-15 parts of bacillus lysinate, 10-20 parts of bacillus licheniformis, 15-25 parts of trichoderma viride, 5-10 parts of candida glabrata and 5-10 parts of yarrowia lipolytica are uniformly mixed according to the weight ratio, and the dosage of the biosurfactant is 1-5 parts.
In a specific embodiment, the culture temperature of the bacillus subtilis, the bacillus lysinate and the bacillus licheniformis during the activation culture in the step A is 30-35 ℃, and the activation culture mediums are beef extract peptone liquid culture mediums; in the step A, the culture temperature of the trichoderma viride, the candida glabrata and the yarrowia lipolytica during the activation culture is 20-25 ℃, and the activation culture mediums are all potato dextrose solid culture mediums.
In a specific embodiment, the acclimatization culture in the step B is carried out in a solid screening culture medium containing sodium chloride, firstly, the strains which are not acclimatized are cultured under the condition that the concentration of the sodium chloride in the solid screening culture medium is 4-6 g/L, and the strains in the last step of the acclimatization culture are cultured under the condition that the concentration of the sodium chloride in the solid screening culture medium is 12-14 g/L.
In a specific embodiment, the culture temperature of the bacillus subtilis, the bacillus lysinate and the bacillus licheniformis during the domestication culture in the step B is 30-35 ℃, and the domestication culture mediums are all beef extract peptone solid culture mediums containing sodium chloride; and C, culturing the trichoderma viride, the candida glabrata and the yarrowia lipolytica during domestication culture in the step B at the temperature of 20-25 ℃, wherein domestication culture media are all potato glucose solid culture media containing sodium chloride.
In a specific embodiment, in the step C liquid fermentation expanded culture, six kinds of bacteria obtained by acclimatizing to a medium with the highest sodium chloride concentration are respectively inoculated into a fermentation tank for liquid fermentation expanded culture.
In a specific embodiment, the concentration of the bacterial liquid of the six bacterial strains in the liquid culture medium inoculated to the step C after the domestication in the step B reaches 2 × 108CFU/m L or more, preferably 5 × 108CFU/m L or more.
In a specific embodiment, in the liquid fermentation expanded culture of the step C, the culture temperature of the bacillus subtilis, the bacillus lysinate and the bacillus licheniformis is 30-35 ℃, and the liquid fermentation expanded culture mediums are all beef extract peptone liquid culture mediums; and C, in the liquid fermentation amplification culture in the step C, the culture temperature of the trichoderma viride, the candida glabrata and the yarrowia lipolytica is 20-25 ℃, and the liquid fermentation amplification culture mediums are all potato glucose liquid culture mediums.
In a specific embodiment, the drying temperature in step D is from-10 ℃ to-50 ℃.
In a specific embodiment, the degrading bacteria agent comprises 20-30 wt% of bacillus subtilis, 10-15 wt% of lysine bacillus, 10-20 wt% of bacillus licheniformis, 15-25 wt% of trichoderma viride, 5-10 wt% of candida glabrata, 5-10 wt% of yarrowia lipolytica and 1-5 wt% of biosurfactant, and the total effective viable bacteria number in the degrading bacteria agent is not lower than 9 × 109CFU/g，The biosurfactant is one or more of sophorolipid, rhamnolipid, seaweed glycolipid and saponin, and preferably the biosurfactant is a mixture of more than three of the sophorolipid, the rhamnolipid, the seaweed glycolipid and the saponin.
The invention also provides a degrading microbial inoculum for treating the kitchen waste, which comprises 20-30 parts of bacillus subtilis, 10-15 parts of lysine bacillus, 10-20 parts of bacillus licheniformis, 15-25 parts of trichoderma viride, 5-10 parts of candida glabrata, 5-10 parts of yarrowia lipolytica and 1-5 parts of a biosurfactant.
In a specific embodiment, the degrading bacterial agent is prepared by mixing dormant microbe dry powder containing six microbes of bacillus subtilis, bacillus lysinate, bacillus licheniformis, trichoderma viride, candida glabrata and yarrowia lipolytica with biosurfactant.
In a specific embodiment, the dormant microorganism dry powder is obtained by performing acclimatization culture and dehydration drying on six strains separately, and the acclimatization culture is a step-by-step acclimatization culture for gradually increasing the concentration of sodium chloride for culture on each microorganism.
Preferably, compared with the degradation efficiency of the degradation microbial inoculum prepared by mixing six directly purchased strains and mixing the six strains with the biosurfactant, the degradation efficiency of the degradation microbial inoculum prepared by using the single strain domesticated and cultured by the method is about doubled. That is to say, if the degrading bacteria agent a obtained by directly mixing six commercially available single bacteria has the degrading efficiency of 50-60% of the degrading efficiency of the degrading bacteria agent b obtained by mixing the single bacteria obtained by gradient acclimation of sodium chloride to the kitchen waste.
In a specific embodiment, the degrading bacterial agent comprises 20-30 wt% of bacillus subtilis, 10-15 wt% of lysine bacillus, 10-20 wt% of bacillus licheniformis, 15-25 wt% of trichoderma viride, 5-10 wt% of candida glabrata, 5-10 wt% of yarrowia lipolytica and 1-5 wt% of biosurfactant.
In a specific embodiment, the degrading bacteriaThe agent is powder, and the total effective viable count is not less than 9 × 109CFU/g。
In a specific embodiment, the biosurfactant is one or more selected from sophorolipids, rhamnolipids, algal glycolipids and saponins, and preferably the biosurfactant is a mixture of three or more thereof.
In a specific embodiment, the biosurfactant is prepared by adding water to 100 parts by weight of sophorolipid 20-25 parts, rhamnolipid 5-10 parts, algal glycolipid 5-10 parts and saponin 1-5 parts, and mixing and dissolving; the mass concentration of the biosurfactant is preferably 45% or more.
In a specific embodiment, the preparation method of the degrading microbial inoculum comprises the following steps: step A, activation culture: firstly, respectively inoculating six bacteria of bacillus subtilis, bacillus lysinate, bacillus licheniformis, trichoderma viride, candida glabrata and yarrowia lipolytica into activation culture media of the six bacteria for activation culture; step B, domestication culture: then, carrying out domestication culture on the six bacteria respectively, wherein the domestication culture is a step-by-step domestication culture for gradually increasing the concentration of sodium chloride for culture on each microorganism; c, liquid fermentation and enlarged culture: respectively inoculating six bacteria obtained by domestication into a fermentation tank of the six bacteria for liquid fermentation and expanded culture; step D, dehydration and drying: dehydrating and drying various single bacteria obtained by amplification culture to prepare dormant microorganism dry powder; step E, mixing uniformly: and D, uniformly mixing the six dormant body microorganism dry powders prepared in the step D, and uniformly mixing the six dormant body microorganism dry powders with a biosurfactant to obtain the degrading microbial inoculum for treating the kitchen waste.
In a specific embodiment, in the step B, domestication culture is carried out in a solid screening culture medium containing sodium chloride, firstly, the strains which are not domesticated are cultured under the condition that the concentration of sodium chloride in the solid screening culture medium is 4-6 g/L, in the last step of domestication culture, the strains are cultured under the condition that the concentration of sodium chloride in the solid screening culture medium is 12-14 g/L, in the step B, the culture temperature of bacillus subtilis, bacillus lysinate and bacillus licheniformis is 30-35 ℃, the domestication culture medium is a beef extract peptone solid culture medium containing sodium chloride, in the step B, the culture temperature of trichoderma viride, candida glabrata and yarrowia lipolytica is 20-25 ℃, and the domestication culture medium is a potato glucose solid culture medium containing sodium chloride.
The invention also provides an application of the degrading microbial inoculum, which comprises the steps of mixing 50-80 g of the degrading microbial inoculum in each ton of kitchen waste in the kitchen waste integrated treatment equipment, controlling the temperature of the kitchen waste degradation treatment to be more than 45 ℃, and carrying out ventilation stirring treatment on the kitchen waste, wherein the temperature of the kitchen waste degradation treatment is preferably controlled to be more than 50 ℃, and the time of the degradation treatment is 3-5 days.
The beneficial effects of the invention at least comprise the following contents:
1. in the degrading microbial agent, bacillus subtilis, bacillus lysinate, bacillus licheniformis, trichoderma viride, candida glabrata and yarrowia lipolytica all have tolerance to the degradation of salt in the kitchen waste, and particularly, the salt-tolerant strains domesticated by increasing the concentration gradient of sodium chloride can better tolerate the high-concentration salt in the kitchen waste. The degrading microbial inoculum provides a solid guarantee for exerting high-efficiency biological activity in high-concentration salt kitchen waste.
2. The bacillus subtilis and the yarrowia lipolytica in the degrading microbial inoculum can degrade grease in kitchen waste, and the bacillus subtilis and the yarrowia lipolytica are compounded to ensure that the grease degrading microbial inoculum has high grease degrading rate.
3. The biosurfactant of the invention not only has the common properties of surfactants such as solubilization, emulsification, wetting, foaming, dispersion, surface tension reduction and the like. Compared with other surfactants produced by chemical synthesis or petroleum refining methods, the biosurfactant also has the advantages of no toxicity, biodegradability, ecological safety, high surface activity and the like, and the biosurfactant also has the following advantages: the application range is wide, and the method can be almost used in various fields; the molecular structure types are various, and part of the molecular structures have a plurality of special functional groups, so that the surface performance is excellent; the biological toxicity is extremely low, the environment is friendly, and the biodegradable performance is 100 percent; is suitable for extreme temperature, pH value and salinity.
4. In general, the degrading microbial inoculum or the degrading microbial inoculum prepared by the method can rapidly decompose macromolecular organic matters such as protein, grease, starch polysaccharide, cellulose, hemicellulose and the like in the kitchen waste, so that the macromolecular organic matters are rapidly stabilized, the degrading efficiency of the kitchen waste is improved, and the space utilization rate of a fermentation bin is further improved. According to the invention, the biological surfactant is added into the degrading microbial inoculum to solubilize, emulsify, wet, disperse and reduce the surface tension of the grease in the kitchen waste, and on the basis, the grease is rapidly decomposed by the bacillus subtilis and the yarrowia lipolytica which have the function of efficiently biodegrading the grease, so that the degrading efficiency of the kitchen waste is improved, and the product quality of the organic fertilizer is improved. Through tests, in the process of treating the kitchen waste by the kitchen waste integrated treatment equipment, the degrading bacteria agent is added in a certain proportion, the oil content can be reduced from 4.5% to 0.3-0.5% within 72 hours, and the degradation rate reaches more than 87%. The result of the germination index of the seeds shows that the addition of a certain proportion of the degrading microbial inoculum can effectively improve the maturity of the materials, improve the fermentation efficiency and shorten the fermentation period. The kitchen waste treatment method is simple and rapid, obvious in effect and high in degradation efficiency, shortens the fermentation time of the kitchen waste, is low in treatment cost, has no biotoxicity, can be biodegraded, does not cause secondary pollution to the environment, and is high in quality of the degraded organic fertilizer product.
Drawings
In the three conical flasks in FIG. 1, the Bacillus subtilis seed solution is obtained after the activation culture of the strains.
Fig. 2 and fig. 3 are both gradient screening culture dishes of bacillus subtilis in the acclimatization culture process, wherein three culture dishes in the gradient screening culture dishes represent 3 parallel tests with the same sodium chloride concentration gradient arrangement, wherein fig. 2 shows the result of 48h culture, and both culture dishes in fig. 3 show the result of 24h culture.
FIG. 4 is a photograph showing the germination of seeds in the treated group using the degrading microbial inoculum according to the present invention, wherein three plates represent three parallel experiments.
Detailed Description
The invention is further illustrated by the following examples and figures, but the scope of protection of the invention is not limited to the following examples.
Example 1
The degrading bacterial agent, the preparation method and the application thereof comprise the following steps:
(1) activated culture of strain
Firstly, respectively inoculating bacillus subtilis, bacillus lysinate and bacillus licheniformis to beef extract peptone liquid culture medium for activation culture, placing the beef extract peptone liquid culture medium in a constant-temperature shaking table with the temperature of 30-35 ℃ and the rotational speed of 180-.
The beef extract peptone liquid culture medium comprises 3g of beef extract, 10g of peptone, 2g of yeast extract, 5g of sodium chloride, 1000m of ultrapure water L, and has a pH value of 7.4-7.6.
Then inoculating Trichoderma viride, Candida glabrata and yarrowia lipolytica strains into a potato glucose culture medium for activation culture, placing the potato glucose culture medium in a constant temperature shaking table with the temperature of 20-25 ℃ and the rotational speed of 180-220rpm for shake culture for 2-4 days until logarithmic growth period, and obtaining activated liquid seed liquid.
The solid culture formula of potato glucose comprises potato 200g, glucose 20g, ultrapure water 1000m L, and pH 7.4-7.6.
For example, the case of Bacillus subtilis seed solution after the seed activation culture is shown in each of three Erlenmeyer flasks in FIG. 1.
(2) Domestication and culture of strain
And respectively inoculating the activated seed liquid into solid screening culture media with the sodium chloride concentration increased in a gradient manner, wherein the initial concentration of the sodium chloride of the unacclimated strain is set to be 5.0 g/L, the sodium chloride concentration is increased by 2.0 g/L each time, and the other four gradients are respectively 7.0 g/L, 9.0 g/L, 11.0 g/L and 13.0 g/L.
In the domestication process, the chloride ion concentration is gradually increased according to the increase of the average colony number per unit, each time the chloride ion concentration is increased by 100 mg/L, after the growth of the bacteria is stable, the chloride ion concentration is gradually increased, and the bacteria are gradually domesticated to obtain the strains.
The domestication culture medium for the bacillus subtilis, the bacillus lysimachiae and the bacillus licheniformis is prepared from 3g of beef extract, 10g of peptone, 2g of yeast extract, 5.0g/7.0g/9.0g/11.0g/13.0g of sodium chloride, 15-20g of agar, 1000m of ultrapure water L and pH7.4-7.6, wherein the seed liquid is taken 1m L respectively, diluted by 200 times with ultrapure water, uniformly coated in each gradient screening culture medium culture dish, numbered respectively, uniformly placed in a constant temperature incubator at 30-35 ℃ for standing culture for 1-2 days, and each gradient of each strain is provided with 3 parallel tests.
The domestication culture medium for Trichoderma viride, Candida glabrata and yarrowia lipolytica comprises 200g of potato, 20g of glucose, 5.0g/7.0g/9.0g/11.0g/13.0g of sodium chloride, 15-20g of agar, 1000m L of ultrapure water and pH7.4-7.6, respectively taking the seed liquid 1m L, diluting with ultrapure water by 200 times, uniformly coating the seed liquid in each gradient screening culture medium culture dish, respectively numbering, uniformly placing in a constant temperature culture box at the temperature of 20-25 ℃ for standing culture for 2-4 days, and setting 3 parallel tests for each gradient of each strain.
FIGS. 2 and 3 are gradient screening culture dishes of Bacillus subtilis in the acclimatization culture process, wherein three culture dishes represent 3 parallel tests with the same sodium chloride concentration gradient arrangement, wherein FIG. 2 shows the result of 48h culture, and both culture dishes in FIG. 3 show the result of 24h culture.
TABLE 1
(3) Counting and analyzing
The number of colonies in the culture medium of each strain gradient screening is shown in table 1, the results in table 1 show that the number of the colonies in the culture dish is obviously reduced along with the increasing of the concentration of the sodium chloride of different strains, which shows that the high concentration of the sodium chloride has obvious inhibition effect on the growth of the microorganism, and the colonies on the culture dish with the concentration of the sodium chloride of 13.0 g/L are taken as the strains for amplification culture through the gradient screening.
(4) Expanding culture
Respectively picking out sodium chloride with concentration of 13.0 g/L for cultureInoculating the bacterial colony on the dish into a liquid culture medium which is the same as the activated culture medium, wherein the concentration of the bacterial liquid is required to reach 2 × 108CFU/m L and above, preferably 5 × 10 colony number8CFU/mL。
(5) Drying at low temperature, and mixing
Drying the culture expanding solution at low temperature to obtain dormant microorganism dry powder. And then uniformly mixing the components according to the weight ratio of 20-30 parts of bacillus subtilis, 10-15 parts of lysine bacillus, 10-20 parts of bacillus licheniformis, 15-25 parts of trichoderma viride, 5-10 parts of candida glabrata and 5-10 parts of yarrowia lipolytica for later use.
(6) Preparation of biosurfactant
25 parts of sophorolipid, 5 parts of rhamnolipid and 5 parts of algal glycolipid with the mass concentration of 45 percent are sequentially added into 65 parts of ultrapure water and are uniformly mixed to prepare the biosurfactant.
(7) Preparation of degrading microbial inoculum for kitchen waste integrated treatment equipment
Uniformly mixing 95-99 parts of the mixed microbial inoculum with 1-5 parts of a biosurfactant to prepare a degrading microbial inoculum for integrated treatment equipment for kitchen waste, controlling the mass water content of the microbial inoculum to be below 3-5%, and controlling the effective viable count to be more than or equal to 9 × 109CFU/g。
(8) Adding degradation microbial inoculum
Adding a degrading microbial inoculum into kitchen waste integrated treatment equipment, mixing the degrading microbial inoculum with kitchen waste, and adding 50-80 g of degrading microbial inoculum per ton of waste; adding degrading bacteria, and fermenting the kitchen waste for 3-5 days (treatment group) by keeping the temperature above 50 ℃ and ventilating and stirring.
(9) Comparative test
The commercial decomposing inoculant of the Rew decomposing accelerator of the Reyuanbi Biotechnology development Limited company of externally purchased wall market has the effective viable count of more than or equal to 200 hundred million/g (namely the viable count of more than or equal to 2 × 10)10CFU/g). Adding a commodity decomposing microbial inoculum into kitchen waste integrated treatment equipment, mixing the kitchen waste with the commodity decomposing microbial inoculum, and adding 50-80 g of the commodity decomposing microbial inoculum per ton of waste; after adding the commercial decomposing microbial inoculum, the kitchen waste is subjected to heat preservation at the temperature of more than 50 ℃ and ventilation stirring fermentation for 3-5 days to complete the fermentation process (comparison group).
(10) Determination of oil content
The degrading bacteria and the commercial decomposition bacteria in a comparative test are respectively added, 10g (dry weight) of organic fertilizer semi-finished products are formed in the fermentation process, the content of grease in the fermentation materials is measured by a Soxhlet extraction method, the Soxhlet extraction method executes the standard of 'method for measuring fat in food' (GB 5009.6-85), the specific measuring method also refers to the standard of 'method for measuring fat in food' (GB 5009.6-85), and each treatment is repeated for 3 times. Table 2 lists the statistics of the determination of oil content in the treatment groups of the degrading microbial inoculum of the invention and the control group using the commercial decomposing microbial inoculum.
TABLE 2
As can be seen from Table 2, after the kitchen waste raw materials are degraded, the oil content is reduced to 0.45%, and the degradation rate exceeds 87%. The degradation rate of the comparison group is only 30%, and the degradation rate of the grease of the treatment group is far higher than that of the comparison group.
(11) Determination of seed germination index
Respectively taking the degrading microbial inoculum and a commercial decomposition microbial inoculum for a comparative test, completing the fermentation process to form 20g (dry weight) of an organic fertilizer semi-finished product, adding 200ml of distilled water, oscillating for 20min, leaching at 30 ℃ for one day and night, filtering supernate by using slow filter paper, and standing filtrate for later use;
respectively spreading one piece of filter paper with corresponding size in a 9cm culture dish, uniformly putting 20 full small vegetable seeds with similar size, sucking 5.0m L leaching liquor into the culture dish by a pipette, and performing a control experiment by using distilled water, wherein each treatment is repeated for 3 times;
placing the culture dish in an incubator with 25 ℃ and 80% humidity for culturing for 24 hours;
the germination rate and root length of the seeds were measured and GI was calculated.
If GI > 60%, the material is considered to be essentially decomposed, and when GI reaches 80% -85%, the material is considered to be completely decomposed and not toxic to plants.
Table 3 shows statistics of seed germination indices in the treatment group with the degrading microbial inoculum of the present invention and the control group with the commercial decomposing microbial inoculum.
TABLE 3
As can be seen from Table 3, the average Germination Index (GI) of the seeds in the treated group reaches 88.5%, which indicates that the material is completely decomposed and has no toxicity to plants, and the application of the material to soil has no toxic or side effect on the growth of the plants. And the average value of the Germination Index (GI) of the seeds in the comparison group is only 64.6 percent, which indicates that the materials are basically decomposed and do not reach the standard of complete decomposition. It can be known through comparative analysis that the comparative group can not reach the complete decomposition standard of the treatment group because the strains in the comparative group are not properly compounded, the efficiency of decomposing the grease is low, the salt has an inhibiting effect on microbial fermentation and other factors, the fermentation efficiency is not high, the grease is not completely degraded, and the fermentation period is not completely decomposed within 3-5 days.
FIG. 4 shows the germination of seeds from the treated group of the present invention using a degrading microbial inoculum, in which three plates represent three replicates.
The foregoing is a more detailed description of the invention in connection with specific preferred embodiments and it is not intended that the invention be limited to these specific details. For those skilled in the art to which the invention pertains, several simple deductions and substitutions can be made without departing from the spirit of the invention, and all shall be considered as belonging to the protection scope of the invention.
Claims (10)
1. A preparation method of a degrading microbial inoculum for treating kitchen waste comprises bacillus subtilis, bacillus lysinate, bacillus licheniformis, trichoderma viride, candida glabrata, yarrowia lipolytica and a biosurfactant; the preparation method comprises the following steps:
step A, activation culture: firstly, respectively inoculating six bacteria of bacillus subtilis, bacillus lysinate, bacillus licheniformis, trichoderma viride, candida glabrata and yarrowia lipolytica into activation culture media of the six bacteria for activation culture;
step B, domestication culture: then, carrying out domestication culture on the six bacteria respectively, wherein the domestication culture is a step-by-step domestication culture for gradually increasing the concentration of sodium chloride for culture on each microorganism;
c, liquid fermentation and enlarged culture: respectively inoculating six bacteria obtained by domestication into a fermentation tank of the six bacteria for liquid fermentation and expanded culture;
step D, dehydration and drying: dehydrating and drying various single bacteria obtained by amplification culture to prepare dormant microorganism dry powder;
step E, mixing uniformly: and D, uniformly mixing the six dormant body microorganism dry powders prepared in the step D, and uniformly mixing the six dormant body microorganism dry powders with a biosurfactant to obtain the degrading microbial inoculum for treating the kitchen waste.
2. The method according to claim 1, wherein the step D comprises mixing the components in a weight ratio of 20-30 parts of Bacillus subtilis, 10-15 parts of Bacillus lysinate, 10-20 parts of Bacillus licheniformis, 15-25 parts of Trichoderma viride, 5-10 parts of Candida glabrata and 5-10 parts of yarrowia lipolytica, and the amount of the biosurfactant is 1-5 parts by weight.
3. The method according to claim 1, wherein the culture temperature of the bacillus subtilis, the bacillus lysinate and the bacillus licheniformis during the activation culture in the step A is 30-35 ℃, and the activation culture mediums are beef extract peptone liquid culture mediums; in the step A, the culture temperature of the trichoderma viride, the candida glabrata and the yarrowia lipolytica during the activation culture is 20-25 ℃, and the activation culture mediums are all potato dextrose solid culture mediums.
4. The method of claim 1, wherein the acclimatization culture in step B is performed in a solid screening culture medium containing sodium chloride, the first unacclimated strain is cultured in the solid screening culture medium at a sodium chloride concentration of 4-6 g/L, and the last step of the acclimatization culture is performed at a sodium chloride concentration of 12-14 g/L.
5. The method according to claim 4, wherein the culture temperature of the bacillus subtilis, the bacillus lysinate and the bacillus licheniformis in the domestication culture in the step B is 30-35 ℃, and the domestication culture mediums are all beef extract peptone solid mediums containing sodium chloride; and C, culturing the trichoderma viride, the candida glabrata and the yarrowia lipolytica during domestication culture in the step B at the temperature of 20-25 ℃, wherein domestication culture media are all potato glucose solid culture media containing sodium chloride.
6. The method according to claim 4, wherein in the step C liquid fermentation scale-up culture, six kinds of bacteria obtained by acclimatizing to a medium having a maximum sodium chloride concentration are respectively inoculated into fermentation tanks thereof for liquid fermentation scale-up culture.
7. The method of claim 1, wherein the concentration of the six bacterial strains in the liquid culture medium inoculated to the step C after acclimatization in the step B is 2 × 108CFU/m L or more, preferably 5 × 108CFU/m L or more.
8. The method according to claim 1, wherein in the step C, the culture temperature of the bacillus subtilis, the bacillus lysinate and the bacillus licheniformis is 30-35 ℃ in the liquid fermentation expanding culture, and the liquid fermentation expanding culture mediums are all beef extract peptone liquid culture mediums; and C, in the liquid fermentation amplification culture in the step C, the culture temperature of the trichoderma viride, the candida glabrata and the yarrowia lipolytica is 20-25 ℃, and the liquid fermentation amplification culture mediums are all potato glucose liquid culture mediums.
9. The method of claim 1, wherein the drying temperature in step D is from-10 ℃ to-50 ℃.
10. The method according to any one of claims 1 to 9, wherein the degrading bacteria agent comprises 20 to 30 wt% of bacillus subtilis, 10 to 15 wt% of lysine bacillus, 10 to 20 wt% of bacillus licheniformis, 15 to 25 wt% of trichoderma viride, 5 to 10 wt% of candida glabrata, 5 to 10 wt% of yarrowia lipolytica and 1 to 5 wt% of biosurfactant, and the total effective viable count in the degrading bacteria agent is not less than 9 × 109CFU/g, the biosurfactant is one or more of sophorolipid, rhamnolipid, seaweed glycolipid and saponin, and preferably the biosurfactant is a mixture of more than three of the sophorolipid, the rhamnolipid, the seaweed glycolipid and the saponin.
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