CN109370944B - Bacillus capable of producing protease and application of bacillus in compost - Google Patents
Bacillus capable of producing protease and application of bacillus in compost Download PDFInfo
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Abstract
The invention discloses a protease-producing bacillus and application thereof in composting, belonging to the technical field of environmental biology. The Bacillus hisashii strain B207 provided by the invention has good sludge protein degradation capability, can be well symbiotic with other strains, and is scientific and reasonable in compounding ratio. The microbial agent containing the strain is used for composting, the time required for the compost to reach high temperature is only 12-24 hours, and after composting for 1.5 days, the degradation rate of protein in the compost raw materials can reach 95.2% -98.9%, so that the composting reaction speed can be effectively increased, the harmless effect of the compost can be improved, and the application prospect is extremely wide.
Description
Technical Field
The invention relates to a protease-producing bacillus and application thereof in composting, belonging to the technical field of environmental biology.
Background
By the end of 2015 6 months, there are 3802 urban sewage treatment plants in China, and the daily capacity of treating domestic sewage reaches 1.6 billion cubic meters. The amount of the generated excess sludge is about 0.3 to 0.5 percent of the sewage treatment amount along with the sewage treatment process, and the amount of the excess sludge generated in China is about 4000 ten thousand tons (according to 80 percent of water content) every day and is continuously increased by 8 to 10 percent of annual growth rate. The excess sludge of sewage treatment contains rich plant nutrient substances, such as 2-7% of nitrogen, 1-5% of phosphorus and 0.1-0.8% of potassium in municipal sludge. The potassium content of the digested sludge is less, but the nitrogen and phosphorus content of the digested sludge is similar to that of the manure. The nitrogen and phosphorus content of the activated sludge is 4-5 times of that of the manure. In addition, the sludge also contains trace elements such as sulfur, iron, calcium, sodium, magnesium, zinc, copper, molybdenum and the like, and abundant organic matters and humus. The sludge compost is utilized, so that the fertilizer efficiency is better, the soil can form a granular structure, the effect of improving the soil is achieved, and the resource utilization of the sludge is realized.
However, a sewage treatment plant produces a large amount of excess sludge every day, and the excess sludge is stacked in a plant area, periodically transported to a refuse landfill for landfill or incinerated for heat supply after the excess sludge is naturally dried and dehydrated. On one hand, the treatment and disposal modes are high in cost, and harmful components in the sludge are easy to cause secondary pollution to the environment in the process. On the other hand, the sludge contains abundant available organic matters, taking protein as an example, the protein content of the residual sludge of the sewage treatment plant in China is 35-60%, and the residual sludge comprises two parts of microbial protein and extracellular polymeric substance protein. The microorganism accounts for more than 40% of the dry weight of the activated sludge and comprises bacteria and microfauna, the bacteria accounts for 90-95% of the dry weight of the microorganism, the protein accounts for 50-60% of the dry weight of the bacteria, and the protein content in the microfauna is nearly 80%. Extracellular proteins account for about 40% of the total sludge protein.
Aiming at the properties and the compositions of the excess sludge, novel degrading bacteria of protein are screened and are compounded with other microbial strains to obtain the compound microbial agent, which has important significance for accelerating the composting process, improving the composting efficiency and improving the composting sanitary environment.
Disclosure of Invention
The first purpose of the invention is to provide a strain of Bacillus hisashii strain B207 producing protease, which is preserved in China center for type culture collection in 7-13.2018 with the preservation number of CCTCC No: M2018473 and the preservation address of China, Wuhan and Wuhan university. The fermentation culture conditions of the strain are as follows: the temperature is 30-60 ℃, the ventilation volume is 0.5-1vvm, the stirring speed is 200-600r/min, the dissolved oxygen is more than 10 percent, and the pH value is 6.5-7.5.
The second purpose of the invention is to provide a microbial agent containing the Bacillus hisashii strain B207, wherein the quantity of the Bacillus hisashii strain B207 in the microbial agent is more than or equal to 3.0 × 108CFU/g。
In one embodiment of the invention, the amount of Bacillus hisashii strain B207 in the microbial agent is more than or equal to 3.0 × 108CFU/mL。
In one embodiment of the invention, the microbial agent further comprises Bacillus fusiformis (Lysinibacillus fusiformis), Pichia kudriavzevii (Pichia kudriavzevii), Bacillus vegicus (Bacillus oleosus) and Bacillus thermolactis strain B209; the Bacillus thermophilus strain B209 is preserved in China center for type culture Collection in 7 months and 13 days in 2018, the preservation number is CCTCC No. M2018474, and the preservation address is China, Wuhan university.
In one embodiment of the present invention, the Lysinibacillus fusiformis (Lysinibacillus fusiformis) may be selected from Lysinibacillus fusiformis ATCC 7055; the Pichia kudriavzevii can be selected from Pichia kudriavzevii ATCC 24210, and the Bacillus vegicus (Bacillus oleosus) can be selected from Bacillus oleosus DSM 9356.
In one embodiment of the invention, the ratio of viable bacteria number of Bacillus hisashii strain B207, Lysinibacillus fusiformis, Pichia kudriavzevii, Bacillus vegicus and Bacillus thermophilus strain B209 in the microbial agent is 700-860:3.5-6:1-2.5:700-860:1600-2200, and the total viable bacteria number of the microbial agent is 1.0 × 109-1.0×1010CFU/mL or CFU/g.
The third purpose of the invention is to provide a method for preparing the microbial agent, which comprises the steps of respectively inoculating Bacillus hisashii strain B207, lysine Bacillus fusiformis, Pichia kudriavzevii, vegetable Bacillus and Bacillus thermolactis strain B209 into a culture medium to obtain fermentation liquor of the Bacillus hisashii strain B207, the lysine Bacillus fusiformis, the Pichia kudriavzevii, the vegetable Bacillus and Bacillus thermolactis strain B209; and mixing the obtained fermentation liquor according to a ratio, and adding a carrier auxiliary material accounting for 0.1-10% of the mass of the fermentation liquor to obtain the liquid microbial agent.
In one embodiment of the present invention, the carrier auxiliary material includes, but is not limited to, one or more of lignite, zeolite, and stone powder.
In one embodiment of the invention, the preparation method of the composite microbial agent comprises the steps of mixing Bacillus hisashii strain B207, lysine Bacillus fusiformis, Pichia kudriavzevii, vegetable Bacillus and Bacillus thermolactis strain B209 fermentation liquor in proportion, adding one or more of crushed straw, carbonized straw and bran, and drying by hot air to obtain the solid composite microbial agent.
In one embodiment of the invention, Bacillus hisashii strain B207, Bacillus fusiformis, Pichia kudriavzevii, Bacillus vegicus and Bacillus thermolactis strain B209 are respectively inoculated into a nutrient broth culture medium subjected to high-temperature sterilization, the inoculation amount is 3-10% (v/v) of the volume of a fermentation culture medium, standing culture is carried out for 20-40h, the culture temperature of the Bacillus fusiformis strain B is 28-37 ℃, the culture temperature of the Bacillus hisashii strain B207, Bacillus fusiformis and Bacillus thermolactis strain B209 is 30-60 ℃, and when the viable count is more than or equal to 1 × 108And (3) obtaining the fermentation liquor of the Bacillus hisashii strain B207, the lysine Bacillus fusiformis, the vegetable Bacillus and the Bacillus thermolactis strain B209 when the concentration is CFU/mL.
In one embodiment of the invention, the nutrient broth is a nutrient broth after autoclaving.
In one embodiment of the invention, the ingredients of the nutrient broth medium comprise 3.0g of beef extract, 10.0g of peptone, 5.0g of sodium chloride and 1000mL of water.
In one embodiment of the invention, Pichia kudriavzevii is inoculated into a potato glucose medium after high-temperature sterilization, the inoculation amount is 3-10% (v/v) of the volume of a fermentation medium, standing culture is carried out for 20-30h, the culture temperature is 20-30 ℃, and when the viable count is more than or equal to 1 × 108And obtaining the fermentation liquor of the Pichia kudriavzevii yeast at CFU/mL.
In one embodiment of the present invention, the potato glucose medium is a potato glucose medium sterilized at a high temperature of 120 ℃.
In one embodiment of the invention, the ingredients of the potato dextrose medium comprise 6.0g of potato extract powder, 20.0g of glucose and 1000mL of water.
The fourth purpose of the invention is to provide the application of the microbial agent.
In one embodiment of the invention, the application is to promote mixed composting of excess sludge and straw in sewage treatment of food plants.
In one embodiment of the invention, the application comprises the following steps:
(1) and (3) mixing the compound microbial agent and the compost raw materials according to the weight ratio of 0.1-0.5: mixing at a ratio of 100;
(2) composting the mixture, measuring the temperature of the pile body, and turning the pile according to the temperature change.
In one embodiment of the present invention, the step (2) composts the mixture, measures the temperature of the compost at 8:00 am and 6:00 pm, turns the compost once a day when the temperature is higher than 55 ℃, starts to decrease as the composting time increases, stops turning when the temperature is lower than 40 ℃, and ends the composting.
The invention also claims the application of the Bacillus hisashii strain B207 in the fields of food, chemical industry, biology and environment.
Has the advantages that:
(1) the invention discloses Bacillus hisashii strain B207 capable of degrading protein. The strain produces protease, the enzyme activity can reach 119.8U/mL after fermentation for 26 hours, and the strain has better protein degradation capability.
(2) The compost composite microbial agent provided by the invention can be used for composting by mixing excess sludge and straws in sewage treatment of food plants, and better realizes resource utilization of compost by mixing the excess sludge and the straws in sewage treatment of the food plants.
(3) After the composite microbial agent provided by the invention is piled for 1.5 days, the degradation rate of protein in the compost raw material can reach 95.2% -98.9%, and the degradation rate of protein in a control experiment is 58.2%.
(4) The microbial agent provided by the invention has obvious advantages in composting, and can effectively shorten the temperature rise time of a compost, so that the time required by the compost to reach the high temperature of 55 ℃ is shortened from 56h of a control experiment to 12-24h, and the composting period is shortened.
(5) The microbial agent provided by the invention has obvious advantages in composting, can effectively improve the highest temperature of a compost, enables the highest temperature of the compost to be improved to 61-67 ℃ from 58 ℃ of a control experiment, and has good composting sanitary conditions.
(6) The microbial agent provided by the invention has obvious advantages in composting and can improve the harmless effect of the composting, the number of faecal coliform bacteria and the death rate of roundworm eggs are respectively 64-90/g and 95.9% -99.0% when the composting is finished, and the number of faecal coliform bacteria and the death rate of roundworm eggs are respectively 210/g and 88.9% in a control experiment.
Biological material preservation
Bacillus hisashii strain B207 has been preserved in China center for type culture Collection in 2018, 7/13.M, with the preservation number of CCTCC No. M2018473, and the preservation address of China, Wuhan university.
Bacillus thermolactis strain B209 is preserved in China center for type culture Collection in 7.13.2018, with the preservation number of CCTCC No. M2018474 and the preservation address of China, Wuhan university.
Detailed Description
(1) The detection method of the protein content comprises the following steps: and (2) adopting a Kjeldahl method for detection, namely digesting the sample by concentrated sulfuric acid to convert organic nitrogen into inorganic ammonium salt under the condition of a catalyst, then converting the ammonium salt into ammonia under the alkaline condition, distilling the ammonia with water vapor and absorbing the ammonia by excessive boric acid solution, and titrating by standard hydrochloric acid to calculate the nitrogen content in the sample so as to obtain the protein content. The degradation rate of the protein is R (%):
R＝[(m1-m2)/m1/*100％
wherein m is1Is the mass (mg) of protein in the starting material, m2Is the mass of protein (mg) in the material after a certain period of composting.
(2) The method for detecting the faecal coliform population comprises the following steps: the detection method (GB/T19524.1-2004) for the number of faecal coliform bacteria in the fertilizer is adopted for detection.
(3) The detection method of the death rate of the roundworm eggs comprises the following steps: the detection method (GB/T19524.2-2004) for the number of faecal coliform bacteria in the fertilizer is adopted for detection. In a specific embodiment, commercial strains of lysenin bacillus fusiformis, pichia kudriavzevii and bacillus vegicus are selected: lysinibacillus fusiformis ATCC 7055, Pichia kudriavzevii ATCC 24210, Bacillus oleosus DSM 9356, but not limited to the above strains.
Example 1 screening of Bacillus hisashii Strain B207
(1) Mixing the residual sludge of the food plant sewage treatment and the corn straws according to the mass ratio of 1.5: 1, inoculating 5% (w/w) of summer rotten leaves in a certain area of Wuxi city of Jiangsu province, uniformly mixing, composting, measuring the temperature of a compost at 8:00 am and 6:00 pm every day, turning the compost once every day when the temperature is higher than 55 ℃, beginning to reduce the temperature of the compost along with the extension of composting time, stopping turning the compost when the temperature is lower than 40 ℃, and sampling;
(2) taking the compost sample prepared in the step (1) as a sample for strain screening, weighing 10g of compost sample, putting the compost sample into a triangular flask filled with 90mL of sterilized water, and shaking for 1h by a 120r/min shaking table to obtain 10-1Diluting, and sequentially making into 10-4、10-5And 10-6Respectively sucking 100 mu L of the three compost sample diluents, uniformly coating the three compost sample diluents on nutrient agar culture media, Gao's first agar culture media and potato agar culture medium plates, respectively putting the three compost sample diluents into an incubator at 45 ℃ for inverted culture for 1-7 days, and then repeatedly scribing and separating until pure strains are obtained;
(3) and (3) culturing each pure strain by adopting a gelatin identification culture medium, if gelatin is liquefied, expressing that protease can be produced, judging the protease producing capability of the strain according to the liquefaction degree of the gelatin, and identifying the strain with the strongest protease producing capability when the liquefaction degree is heavier and the protease producing capability is stronger.
Example 2 identification of Bacillus hisashii Strain B207
After being cultured on a nutrient agar medium plate at 55 ℃ overnight, a beige flat and round colony with clear edge is formed, and is observed by an optical microscope to be gram-positive, arranged in single or paired cells, rod-shaped, spore-forming and acid-producing by utilizing carbohydrate.
Taxonomic molecular identification is carried out by adopting 16SrDNA sequencing, the sequencing result (1493bp) is subjected to Blast comparison in NCBI, and the similarity of the 16SrDNA base sequence and the Bacillus hishii (Bacillus hishii strain) is the highest and is 99%.
By combining the morphological characteristics, physiological and biochemical characteristics and hereditary characteristics, the protease-producing strain can be identified as Bacillus hishii strain (Bacillus hishii strain), named as Bacillus hishii B207 and preserved in China center for type culture collection in 7-13 months in 2018, with the strain preservation number being CCTCC No. M2018473.
Inoculating the strain to a nutrient agar culture medium plate, culturing at 45 ℃ for 26h, then inoculating to a nutrient broth culture medium, culturing at 45 ℃ for 26h, centrifuging the fermentation liquor, and determining the activity of protease by adopting a Folin phenol method, wherein the activity of the protease is determined to be 119.8U/mL.
Mixing the residual sludge of food plant sewage treatment and corn straws according to the weight ratio of 1.5: 1, mixing the materials in proportion, wherein the water content of the mixed materials is 55 percent, the C/N ratio is 30, uniformly mixing 0.3 percent (by mass) of fermentation liquor of Bacillus hisashii strain B207 with the materials, composting, measuring the temperature of two heaps at 8:00 am and 6:00 pm every day, turning the heaps once every day when the temperature is higher than 55 ℃, beginning to reduce the temperature of the heaps along with the extension of composting time, stopping turning the heaps when the temperature is lower than 40 ℃, and sampling and measuring. The results show that the protein content of the feedstock was reduced by 90.2% after 1.5 days of composting.
Example 3 Bacillus hisashii Strain B207 activation and culture
Bacillus hisashii strain B207 was inoculated on nutrient agar medium plates and cultured at 45 ℃ for 26 h. The nutrient agar culture medium comprises 3.0g of beef extract, 10.0g of peptone and 5.0g of sodium chloride.
Inoculating the activated strain into a 250mL shaking flask filled with 100mL liquid culture medium, and performing shaking culture at 45 ℃ at 160r/min for 26 h. The shake flask culture medium comprises 3.0g of beef extract, 10.0g of peptone and 5.0g of sodium chloride, and the pH value is 7.0.
Inoculating the shake flask culture solution into a 3L fermentation tank according to the volume ratio of 3-10%, stirring at the rotation speed of 200-600r/min, ventilation of 0.5-1.0vvm and dissolved oxygen of more than 10%, and performing fermentation culture at 30-60 ℃ for 20-30 h. The culture medium of the fermentation tank comprises 3.0g of beef extract, 10.0g of peptone, 30g of sodium acetate, 2.0g of potassium dihydrogen phosphate and 5.0g of sodium chloride, and the pH value is 7.0.
Example 4 preparation of liquid Complex microbial agent containing Bacillus hisashii strain B207
The compost composite microbial agent is prepared by compounding Bacillus hisashii strain B207 fermentation liquor and other strain fermentation liquor, and the preparation method comprises the following steps:
(1) fermentation liquor of Bacillus hisashii strain B207, fusiform lysine Bacillus, vegetable Bacillus and Bacillus thermolactis strain B209 is prepared by respectively inoculating Bacillus hisashii strain B207, fusiform lysine Bacillus, vegetable Bacillus and Bacillus thermolactis strain B209 into a nutrient broth culture medium subjected to high-temperature sterilization, wherein the inoculation amount is 3-10% (v/v) of the volume of the fermentation culture medium, the culture temperature of the fusiform lysine Bacillus is 28-37 ℃, the culture temperature of the Bacillus hisashii strain B207, vegetable Bacillus and Bacillus thermolactis strain B209 is 30-60 ℃, and when the number of viable bacteria is more than or equal to 1 × 108Obtaining fermentation liquor of the Bacillus hisashii strain B207, the lysine Bacillus fusiformis, the vegetable Bacillus and the Bacillus thermolactis strain B209 when the concentration is CFU/mL;
(2) the fermentation liquid of the Pichia kudriavzevii is prepared by inoculating the Pichia kudriavzevii into a potato glucose agar culture medium after high-temperature sterilization, wherein the inoculation amount is 3-10% (v/v) of the volume of the fermentation culture medium, the culture time is 20-30h, the culture temperature is 20-30 ℃, and when the viable count is more than or equal to 1 × 108Obtaining fermentation liquor of the Pichia kudriavzevii yeast when CFU/mL;
(3) mixing the obtained fermentation liquor of Bacillus hisashii strain B207, lysine Bacillus fusiformis, Pichia kudriavzevii, vegetable Bacillus and Bacillus thermolactis strain B209 according to a certain proportion to ensure that the strains liveThe ratio of the number of bacteria is 700-860:3.5-6:1-2.5:700-860:1600-2200, one or more of lignite, zeolite and stone powder which are commercially available carrier auxiliary materials and are 0.1-10% of the mass of the fermentation liquor are added, and the total number of the viable bacteria is 1.0 × 109～1.0×1010CFU/mL liquid compound microbial agent.
Example 5 preparation of solid Complex microbial preparation containing Bacillus hisashii strain B207
Mixing the obtained fermentation liquor of Bacillus hisashii strain B207, the obtained fermentation liquor of lysine Bacillus fusiformis, the obtained fermentation liquor of Pichia kudriavzevii, the obtained vegetable Bacillus and the obtained fermentation liquor of Bacillus thermolactis strain B209 according to a certain proportion, adding one or more of commercially available crushed straws, carbonized straws and wheat bran, and drying by hot air to obtain the solid-state composite microbial agent containing the Bacillus hisashii strain B207.
Example 6 application of composite microbial Agents for composting
Mixing the residual sludge of food plant sewage treatment and corn straws according to the weight ratio of 1.5: 1, mixing the mixture according to the mass ratio of 0.3: 100, the addition amount of the microbial inoculum is 0.3 percent, composting is carried out after uniform mixing, the temperature of two heaps is measured at 8:00 in the morning and 6:00 in the afternoon every day, when the temperature is higher than 55 ℃, the heaps are turned once every day, the temperature of the heaps begins to drop along with the extension of composting time, when the temperature is lower than 40 ℃, the turning is stopped, sampling is carried out, and various quality indexes of the compost are measured. The result shows that the time required for the pile body to reach the highest temperature is 15h, and the highest temperature is 66 ℃; after being stacked for 1.5 days, the protein degradation rate is 95.9 percent; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 17.6, and the number of faecal coliform bacteria and the death rate of roundworm eggs are 77/g and 98.2% respectively.
Example 7 use of compost composite microbial Agents
Mixing the residual sludge of food plant sewage treatment and corn straws according to the weight ratio of 2.5: 1, mixing the mixture according to a mass ratio of 0.5 to 0.5, wherein the water content of the mixture is 55 percent, the C/N ratio is 25, and the ratio of viable bacteria of Bacillus hisashii strain B207, lysine Bacillus fusiformis, Pichia kudriavzevii, Bacillus vegicus and Bacillus thermolactis strain B209 is 700:3.5:1:700: 1600: 100, the addition amount of the microbial inoculum is 0.5 percent, composting is carried out after uniform mixing, the temperature of two heaps is measured at 8:00 in the morning and 6:00 in the afternoon every day, when the temperature is higher than 55 ℃, the heaps are turned once every day, the temperature of the heaps begins to drop along with the extension of composting time, when the temperature is lower than 40 ℃, the turning is stopped, sampling is carried out, and various quality indexes of the compost are measured. The result shows that the temperature of the pile body is rapidly increased, the time required for reaching the highest temperature is 24 hours, and the highest temperature is 61 ℃; after being stacked for 1.5 days, the protein degradation rate is 95.2 percent; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 18.1, and the number of faecal coliform bacteria and the death rate of roundworm eggs are 89/g and 95.9 percent respectively.
Example 8 use of compost composite microbial Agents
Mixing the residual sludge of food plant sewage treatment and corn straws according to the weight ratio of 2.5: 1, mixing the mixture according to a mass ratio of 0.5: 100, the addition amount of the microbial inoculum is 0.5 percent, composting is carried out after uniform mixing, the temperature of two heaps is measured at 8:00 in the morning and 6:00 in the afternoon every day, when the temperature is higher than 55 ℃, the heaps are turned once every day, the temperature of the heaps begins to drop along with the extension of composting time, when the temperature is lower than 40 ℃, the turning is stopped, sampling is carried out, and various quality indexes of the compost are measured. The result shows that the temperature of the pile body rises rapidly, the time required for reaching the highest temperature is 12 hours, and the highest temperature is 67 ℃; after being stacked for 1.5 days, the protein degradation rate is 98.9 percent; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 12.6, and the number of faecal coliform bacteria and the death rate of roundworm eggs are respectively 64/g and 99.0 percent.
Example 9 use of compost composite microbial Agents
Mixing the residual sludge of food plant sewage treatment and corn straws according to the weight ratio of 2.0: 1, mixing the materials in a ratio of 55 percent of water content and 27 percent of C/N, wherein the solid compound microbial inoculum and the mixed materials in a mass ratio of 860:6:2.5:860:2200 of viable count of Bacillus hisashii strain B207, fusiform lysine Bacillus, Pichia kudriavzevii, vegetable Bacillus and Bacillus thermolactis strain B209 are mixed according to a mass ratio of 0.1: 100, the addition amount of the microbial inoculum is 0.1 percent, composting is carried out after uniform mixing, the temperature of two heaps is measured at 8:00 in the morning and 6:00 in the afternoon every day, when the temperature is higher than 55 ℃, the heaps are turned once every day, the temperature of the heaps begins to drop along with the extension of composting time, when the temperature is lower than 40 ℃, the turning is stopped, sampling is carried out, and various quality indexes of the compost are measured. The result shows that the temperature of the pile body is rapidly increased, the time required for reaching the highest temperature is 24 hours, and the highest temperature is 61 ℃; after being stacked for 1.5 days, the protein degradation rate is 95.2 percent; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 18.9, and the number of faecal coliform bacteria and the death rate of roundworm eggs are respectively 90/g and 95.9 percent.
Comparative example 1
Mixing the residual sludge of food plant sewage treatment and corn straws according to the weight ratio of 1.5: 1, mixing the mixture according to the mass ratio of 0.3: 100, and the adding amount of the microbial inoculum is 0.3 percent, wherein the ratio of the viable count of the lysine Bacillus fusiformis, the pichia kudriavzevii, the Bacillus vegicus and the Bacillus thermolactis strain B209 is 4:1.8:780:2000, composting is carried out after uniform mixing, the temperature of two heaps is measured at 8:00 am and 6:00 pm every day, when the temperature is higher than 55 ℃, the heaps are turned once every day, the temperature of the heaps begins to decrease along with the extension of composting time, when the temperature is lower than 40 ℃, the turning is stopped, sampling is carried out, and various quality indexes of the compost are measured. The result shows that the time required for the pile body to reach the highest temperature is 56 hours, and the highest temperature is 58 ℃; after being stacked for 1.5 days, the protein degradation rate is 58.2 percent; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 21.5, and the number of faecal coliform bacteria and the death rate of roundworm eggs are 210/g and 88.9 percent respectively.
Comparative example 2
The specific implementation manner is the same as that in example 6, except that the viable bacteria ratio of Bacillus hisashii strain B207, lysine Bacillus fusiformis, Pichia kudriavzevii, Bacillus vegicus and Bacillus thermolactisstrain B209 is different, and is specifically shown in Table 1. The result shows that the time required for the pile body to reach the highest temperature is prolonged to 45-52h, and the highest temperature is 54-56 ℃; after being piled for 1.5 days, the protein degradation rate is only 52.1 to 62.4 percent; when the composting is finished, the carbon-nitrogen ratio of the mixed material is 20.0-23.4, and the number of faecal coliform and the death rate of ascaris ova are 670/g and 66.8-79.6% respectively.
TABLE 1
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. A strain of Bacillus thuringiensis (Bacillus hisashii) B207 producing protease is preserved in China center for type culture collection (CCTCC No: M2018473) in 13.7.2018, and the preservation address is China, Wuhan university.
2. A microbial agent containing Bacillus thuringiensis (Bacillus hisashii) B207 of claim 1, wherein the amount of Bacillus thuringiensis B207 in the microbial agent is more than or equal to 3.0 × 108CFU/g or CFU/mL.
3. The microbial agent according to claim 2, further comprising Bacillus fusiformis (Lysinibacillus fusiformis), Pichia kudriavzevii (Pichia kudriavzevii), Bacillus vegicus (Bacillus oleosus) and Bacillus thermolactis strain B209; the Bacillus thermophilus strain B209 is preserved in China center for type culture Collection in 7 months and 13 days in 2018, the preservation number is CCTCC No. M2018474, and the preservation address is China, Wuhan university.
4. The microbial agent according to claim 3, wherein the ratio of viable bacteria count of Bacillus flexus B207, Bacillus fusiformis, Pichia kudriavzevii, Bacillus vegicus and Bacillus thermolactisstrain B209 in the microbial agent is 700-860:3.5-6:1-2.5:700-860:1600-2200, and the total viable bacteria count of the microbial agent is 1.0 × 109-1.0×1010CFU/mL or CFU/g.
5. The microbial agent according to claim 4, which is prepared by the following steps: respectively inoculating the B207, the Lysinibacillus fusiformis, the Pichia kudriavzevii, the vegetable Bacillus and the Bacillus thermolactis strain B209 into a culture medium to obtain fermentation liquor of the B207, the Lysinibacillus fusiformis, the Pichia kudriavzevii, the vegetable Bacillus and the Bacillus thermolactis strain B209; mixing the obtained fermentation liquor in proportion, and adding a carrier auxiliary material to obtain a liquid microbial agent; the carrier auxiliary materials include but are not limited to one or more of lignite, zeolite and stone powder.
6. The microbial agent according to claim 4, which is prepared by the following steps: mixing Bacillus village B207, lysine bacillus fusiformis, Pichia kudriavzevii, vegetable bacillus and Bacillus thermophilus strain B209 fermentation liquor in proportion, adding one or more of crushed straw, carbonized straw and bran, and drying by hot air to obtain the solid compound microbial agent.
7. Use of Bacillus outflow (Bacillus hisashii) B207 according to claim 1 for environmental treatment of excess sludge.
8. A method for promoting mixed composting of excess sludge and straw in sewage treatment of food factories, which is characterized in that composting is carried out by using a microbial agent containing Bacillus villagens B207 of claim 1.
9. The method of claim 8, comprising the steps of:
(1) mixing the microbial agent as claimed in any one of claims 2 to 6 with compost raw materials according to the ratio of 0.1-0.5: mixing at a ratio of 100;
(2) composting the mixture, measuring the temperature of the pile body, and turning the pile according to the temperature change.
10. The method according to claim 9, wherein the step (2) is composting the mixture by measuring a temperature of the compost 1 to 2 times each day in the morning and afternoon; turning the pile once every day when the temperature is higher than 55 ℃; and (3) the temperature of the stack body begins to fall along with the extension of the stacking time, and when the temperature is lower than 40 ℃, the stack turning is stopped, and the stacking is finished.
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