CN113817641B - Decomposed leaven and application thereof - Google Patents
Decomposed leaven and application thereof Download PDFInfo
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- CN113817641B CN113817641B CN202111144960.1A CN202111144960A CN113817641B CN 113817641 B CN113817641 B CN 113817641B CN 202111144960 A CN202111144960 A CN 202111144960A CN 113817641 B CN113817641 B CN 113817641B
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- sludge
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- pseudomonas
- aerobic fermentation
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05C—NITROGENOUS FERTILISERS
- C05C9/00—Fertilisers containing urea or urea compounds
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/80—Separation, elimination or disposal of harmful substances during the treatment
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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- Chemical Kinetics & Catalysis (AREA)
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Abstract
The invention relates to the technical field of leavening agents, and discloses Pseudomonas (Pseudomonas sp.) LZ4-5, which has the preservation number as follows: CGMCC NO.21761. The invention also discloses a decomposed leaven, which comprises Pseudomonas (Pseudomonas sp.) LZ4-5. The invention also discloses application of Pseudomonas sp LZ4-5 in preparation of the sludge and mussel shell combined film-coated aerobic fermentation decomposed fertilizer. The invention also discloses a sludge and mussel shell combined film-covering aerobic fermentation decomposed fertilizer prepared by applying the decomposed leaven. Adding a proper amount of decomposed leaven into the total fermented material for aerobic fermentation and decomposition of the combined coated sludge and mussel shells, so that organic matters and mussel shell powder in the sludge can be effectively degraded, and decomposed organic fertilizer completely meeting the index requirements of products such as organic fertilizer and the like is obtained; meanwhile, the problem of recycling sludge in the Zhoushan city and the problem of the ecological environment of the shells of the mussels in the Zhoushan city are solved, and a new method is provided for recycling the shells of the mussels.
Description
Technical Field
The invention relates to the technical field of leavening agents, in particular to a decomposing leavening agent and application thereof.
Background
With the increasing importance of our country on environmental protection and the great investment on sewage treatment, the town sewage is effectively collected and treated, and a large amount of municipal sludge is generated in the process. The sludge contains various toxic substances such as pathogens, parasites, heavy metals, refractory organic matters, carcinogenic organic matters and the like, and can cause serious environmental pollution. Meanwhile, the town sludge is rich in elements such as N, P, K, ca and the like, and the sludge after harmless treatment can improve the physical and chemical properties of soil and increase the crop yield, so that the method has a certain agricultural application value. The sludge treatment comprises sludge treatment and sludge treatment, and aims to perform harmless treatment on the sludge and avoid causing harm to the environment. The sludge treatment process is a process of sludge reduction, harmlessness and stabilization, and the sludge disposal process is a process of placing sludge in a natural environment or recycling.
In terms of sludge treatment, the selection of a sludge treatment mode depends on local conditions, and in the aspect of land utilization, the investment of sludge land utilization is low, the energy consumption is low, but toxic and harmful substances such as heavy metals in sludge are limiting factors; in the aspect of sanitary landfill, sludge may cause groundwater pollution; in the aspect of building material utilization, the odor has the problem of influencing the living environment.
According to the research, the concentration of the odor in the sludge treatment system is 2 to 3 times of that in the sewage treatment system. The cause of odor is reduction of sulfate to H 2 S, the organic matters are decomposed into intermediate products such as alcohol, aldehyde and the like, and in addition, when sewage flows through the grating, large-particle floating objects contain a large amount of organic matters which are intercepted by the grating to cause pollutant accumulation, and various malodorous gases are generated by fermentation. Therefore, the sludge has odor, and there is a need to solve this problem in the utilization treatment process.
At present, the deodorization method comprises a physical method, a chemical method and a biological method.
The active carbon adsorption method is the most widely applied physical deodorization method at present, and can effectively remove a plurality of substances such as indole, sulfide and the like. However, the cost of the activated carbon material is high, the adsorption capacity is limited, the saturation point is difficult to master, and the adsorption capacity of the activated carbon is greatly influenced by the factors such as the components, the temperature, the humidity, the dust content and the like of the malodorous gas.
The chemical absorption method, also called chemical washing method, wet absorption oxidation method, etc., is a method of deodorizing by using an absorption tower as a reaction device in combination with the principles of acid-base neutralization reaction and oxidation reaction, and is the most common application method in the chemical deodorization method. The chemical absorption method has the advantages of strong operation flexibility and can reach the optimal deodorization reaction condition by controlling the adding amount and the adding speed of the liquid medicine. The limitations are as follows: the chemical absorption method needs to add a series of chemical reagents, and generates corrosion phenomena on sewage treatment structures such as a deodorization device and each pipeline; the strong acid or strong base needs to consider the operation safety factor when in use, and the waste liquid generated after absorption is easy to generate secondary pollution.
In the technology of deodorizing urban sewage and sludge treatment plants by a biological method, the most mature and commonly used method of the current practical engineering is that a microorganism packing layer is also used as a main core component of the biological deodorization method, which not only provides growth attachment points for microorganisms, but also provides sufficient nutrient substances for the microorganisms to ensure the activity of the microorganisms, wherein the microorganism packing layer comprises a carbon source and trace elements, and in addition, the packing layer needs to keep the relative stability of the microorganism growth environment, including humidity, oxygen content and pH value. The common fillers comprise bioactive fillers such as hay, dried withered bark, fruit shell, crushed stone, peat and the like, thereby achieving the aim of deodorization. Has the advantages of low energy consumption, flexible investment, almost no secondary pollution and the like, and has good application prospect in the field of sewage deodorization in the future. The technical advantages of the film-covered aerobic fermentation are as follows: (1) By combining biotechnology with molecular selection membrane materials, the method aims at solving the problems of large investment in fixed assets, long treatment period, high cost, low harmful bacteria killing rate, single use, poor weather condition adaptability and the like of the traditional organic solid waste treatment method, and achieves the aim of quickly and efficiently treating the organic solid waste at low cost throughout the year through the composite microbial bacteria and the special molecular selection membrane. (2) The movable and portable characteristics of the fermentation technology effectively improve the defects of insufficient movable flexibility, limited increase and decrease of fermentation capacity and the like existing in the conventional method for treating solid wastes. (3) The fermentation technology covers the molecular selection film on the stack body and compacts the film to form an air chamber, and the molecular selection film has the functions of ventilation, moisture permeability and heat preservation, so that the normal volatilization of water vapor of the stack body can be ensured, and a certain humidity and temperature of the stack body can be maintained; the continuous processing capacity of 12 months can be kept in northern areas, and the method is not influenced by regions and climates, so that the stability of processing capacity is ensured; the southern area is not influenced by large air humidity and the like, and the unidirectional water permeation of the molecular film is beneficial to the volatilization of water in the stack body in the fermentation process. (4) The membrane material adopted by the fermentation technology only can allow water vapor and air to permeate out; dust, bacteria and macromolecular peculiar smell substances in the pile body cannot leak out, so that the environment is protected; the outside rain and snow cannot permeate, so that the outdoor operation condition is provided; light weight, good flexibility, repeated use, aging resistance and long service life.
Mussel culture is an industry which is rapidly growing in the world, and at present, china is the country with the largest yield of mussels in the world. In 2012, the annual yield of the mussels in China reaches 50 ten thousand t. However, if mussel shells accounting for 1/3 of the weight of the mussels are discarded, about 15 to 18 million of mussel shells can be produced in 1 year. Such a lot of discarded mussel shells accumulate or arrive in nearby sea areas or beaches, can cause serious environmental pollution, and the expansion of the resource utilization approach of discarded mussel shells becomes an urgent problem to be solved.
Disclosure of Invention
It is an object of the present invention to address at least the above problems and to provide at least the advantages described hereinafter.
To achieve these objects and other advantages in accordance with the present invention, there is provided a pseudomonad (Pseudomonas sp.) LZ4-5 having a deposit number of: CGMCC NO.21761.
A decomposing leaven comprises Pseudomonas sp LZ4-5.
Preferably, the method further comprises the following steps: at least one of Bacillus subtilis or yeast.
Preferably, the decomposing leaven comprises 1 part of bacillus subtilis and 1 part of yeast and 2 parts of Pseudomonas (Pseudomonas sp.) LZ4-5 by weight. Wherein, the effective viable count of the LZ4-5 of Pseudomonas (Pseudomonas sp.) is 100-200 hundred million/g, the effective viable count of the Bacillus subtilis is 100-200 hundred million/g, and the effective viable count of the microzyme is 100-200 hundred million/g.
An application of Pseudomonas (Pseudomonas sp.) LZ4-5 in preparing sludge and mussel shell combined film-coated aerobic fermentation decomposed fertilizer.
An application of a decomposed starter in preparing a sludge and mussel shell combined film-covered aerobic fermentation decomposed fertilizer.
Preferably, the leavening agent accounts for 0.3 to 0.4 percent of the total fermentation raw materials by weight.
Preferably, the total fermentation raw material also comprises:
38-40% of sludge, 13-15% of rice hull powder and 12-14% of mushroom dregs; 30 to 36 percent of shell powder and 0.6 to 0.7 percent of fermentation nitrogen source.
Preferably, the water content of the sludge is 80-85%, the water content of the rice hull powder is 15-20%, the water content of the mushroom dregs is 35-40%, the shell powder comprises a fine shell powder with the particle size of 40-60 meshes and a coarse shell powder with the particle size of 2-5 meshes, and the weight ratio of the fine shell powder to the coarse shell powder is 10.
A sludge and mussel shell combined film-covering aerobic fermentation decomposed fertilizer prepared by applying a decomposed leavening agent.
The invention at least comprises the following beneficial effects:
the invention provides Pseudomonas (Pseudomonas sp.) LZ4-5 with the preservation number as follows: CGMCC NO.21761 can effectively control the odor in the combined film-covering aerobic fermentation process of the sludge and the mussel shell, and has obvious odor only in the early fermentation stage within 1 meter of the distance from the pile body, and no unpleasant odor is smelled in the middle and later fermentation stages or at a distance of more than 10 meters.
The decomposing leaven provided by the invention can be applied to preparing a combined film-coated aerobic fermentation decomposing fertilizer of sludge and mussel shells, and a proper amount of decomposing leaven can effectively degrade organic matters and mussel shell powder in the sludge to obtain the decomposing organic fertilizer which completely meets the index requirements of products such as organic fertilizers; meanwhile, the problem of recycling sludge in the Zhoushan city and the problem of the ecological environment of the shells of the mussels in the Zhoushan city are solved, and a new method is provided for recycling the shells of the mussels.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Drawings
FIG. 1 is a diagram of a developmental tree analysis of Pseudomonas sp LZ4-5 according to the present invention;
FIG. 2 is a smell change curve of an aerobic fermentation pile in the sludge and mussel shell combined film-covering aerobic fermentation of the invention;
FIG. 3 is a temperature change curve of an aerobic fermentation pile in the sludge and mussel shell combined film-covering aerobic fermentation of the invention;
FIG. 4 is a water content change curve of an aerobic fermentation pile in the sludge and mussel shell combined film-covering aerobic fermentation of the invention;
FIG. 5 is a curve showing the change of organic matter in the aerobic fermentation pile in the combined membrane-covering aerobic fermentation of sludge and mussel shells according to the present invention;
FIG. 6 is a pH change curve of an aerobic fermentation pile in the combined membrane-covering aerobic fermentation of sludge and mussel shells according to the present invention;
FIG. 7 is a variation curve of the carbon-nitrogen ratio of an aerobic fermentation pile in the sludge and mussel shell combined film-covering aerobic fermentation according to the invention;
FIG. 8 is a graph showing the change of nutrients in an aerobic fermentation pile in the combined membrane-covering aerobic fermentation of sludge and mussel shells according to the present invention.
Detailed Description
The present invention is described in further detail below with reference to the attached drawings so that those skilled in the art can implement the invention by referring to the description text.
It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or combinations thereof.
Example 1
Screening and characterization of Pseudomonas sp LZ4-5
1.1 screening method
(1) Sample collection
Collecting a sediment sample: collected in Laizhou gulf, and used at 0.05m 2 Stainless steel GrayO'The Hara box dredger samples seawater sediments were collected at selected survey stations, samples were removed of potential contamination of the surface, and samples were collected about 5cm below the seabed surface of the sediments using sterile 60mL syringes (luer-free tips) and immediately transferred to sterile self-bags for cryopreservation. And (4) preserving the sample at low temperature by using dry ice on a sampling ship, and separating sulfur-oxidizing bacteria after returning to a laboratory.
(2) Separation, purification and preservation of sulfur oxidizing bacteria
Screening and culturing: taking 1g sediment sample, adding sterilized seawater, diluting respectively 10 -1 ，10 -2 ，10 -3 ，10 -4 ， 10 -5 Then, 100ml of the diluted seawater-sediment mixture was aspirated and spread on the sodium thiosulfate solid and sodium sulfide solid medium. And (3) culturing the coated plate in an incubator at 16 ℃, observing the growth condition of bacterial colonies on the plate every day, and inoculating the bacterial strains to a new plate in time for purification when bacterial colonies are formed to show that the bacterial strains have the capacity of degrading sodium thiosulfate.
Purifying sulfur oxidizing bacteria: selecting a single colony formed in the screening culture medium into a newly prepared sodium thiosulfate solid culture medium by using an inoculating ring, purifying by adopting a three-section scribing method, then transferring into a 16 ℃ incubator for culture, and observing the growth condition of the colony on the surface of the culture medium every day. After a single colony is formed, the purification is repeated for one to two times, and the sulfur oxidizing bacteria are preserved after the culture medium is free from mixed bacteria.
Preserving sulfur oxidizing bacteria: and taking a sterilized glycerol tube, and adding 1.5mL of sterilized seawater glycerol. And scraping a proper amount of thalli by using a sterilized 1mL gun head, transferring the thalli into a glycerin pipe, and lightly blowing and beating to ensure that the thalli are uniformly distributed in the seawater glycerin. Two tubes of each strain were stored, marked, placed in a cryopreservation box, and frozen in a-80 ℃ freezer for later use.
1.2 characterization
1.2.1 Biochemical and morphological characterization of Pseudomonas sp LZ4-5, which indicated that Pseudomonas sp LZ4-5 was white translucent with smooth edges, a gram-negative bacterium.
1.2.2 sequence homology analysis of 16S rDNA on Pseudomonas sp LZ4-5
The 16S rDNA fragment of the obtained Pseudomonas (Pseudomonas sp.) LZ4-5 was amplified by colony PCR. The 16S rRNA gene fragment is amplified, cloned and sequenced, and the result shows that the polynucleotide sequence of the 16S rDNA of the Pseudomonas (Pseudomonas sp.) LZ4-5 is shown as SEQ ID NO. 1. Sequence alignment of the 16SrDNA of Pseudomonas sp LZ4-5 is shown in FIG. 1 for a phylogenetic tree analysis of the strain.
Pseudomonas sp LZ4-5 has been deposited at 29.01.2021 in China general microbiological culture Collection center, accession number: xilu No.1 Hospital No. 3, beijing, chaoyang, beicheng. The preservation number is: CGMCC NO.21761.
Example 2
2.1 culture method of strains
1) Adopting 2216E culture medium to culture (purchasing finished product culture medium) LZ4-5 strain, and culturing at 30 deg.C.
2.2 analysis of intermediate metabolites during sodium sulfide degradation
Transferring the strain to be detected to a flat plate with sodium thiosulfate as a substrate;
and (3) inoculating the purified degrading strain in 5% of inoculum size back to an inorganic salt basic culture solution with thiosulfate as a substrate to verify whether the degrading strain has degrading capability, and if turbidity appears, using the strain solution for the next step of operation.
Inoculating 15mL of bacterial liquid (when the bacterial liquid OD600nm =0.22, namely the bacterial liquid concentration is about 0.44 x10 ^9 cfu/mL), culturing in a shaking table at 30 ℃ and 160r/min, periodically sampling in the culturing process (sampling from the shaking table and processing every 12 hours for determination, placing the inoculated conical flask in an ultra-clean workbench for standing and precipitating for 3min, then using a pipette to respectively take 1mL of culture liquid into a 1.5mL sterilized centrifuge tube for determining the OD of the culture liquid at each time point 600 . Sucking the residual liquid with a pipette into a sterilized 50mL centrifuge tube, centrifuging at 5000rpm for 10min, collecting the supernatant, vacuum filtering with 0.45um filter membrane, and vacuum filtering to obtain liquid for measuringThe pH of the culture broth and the concentration of (sulfate, sulfite, thiosulfate) in the liquid phase over time at the time points were compared with a non-inoculated medium.
2.3 detection method
Concentration of sodium thiosulfate: the iodometry procedure was as follows:
(1) Preparing diluted solution, placing 1ml of culture solution in iodine measuring flask, and diluting with distilled water to 50ml (50 times dilution)
(2) Adding 2ml of 17.5mol/L acetic acid and 1ml of 1% starch solution into an iodine measuring bottle
(3) Titrating with 0.01mol/L iodine standard solution until non-disappearing blue color appears, and calculating S by using the iodine amount consumed by titration 2 O 3 2- The content of (b).
(4) A blank test was conducted by replacing the culture medium with 1ml of distilled water.
Calculating the formula: y = (V-V0) C M X1000/50
Wherein: y is S contained 2 O 3 2- The amount of (B) is in g/L
C is the concentration of the titrated iodine standard solution (0.01 mol/L)
M is S 2 O 3 2- Molecular weight of (2) (112 g/mol)
V is the volume of iodine solution consumed by titrating the sample
V0 is the volume of iodine solution consumed for titrating distilled water
I 2 +2S 2 O 3 2- →2I - +S 4 O 6 2-
The calculated sulfur degradation rate of the strain is as follows: 94.2 percent of
Example 3
3.1 fermentation raw material ratio
The list of raw materials is given in Table 3-1 below.
TABLE 3-1 List of aerobic fermentation raw materials of shell powder
Other supporting conditions required by sludge and mussel shell combined film-covering aerobic fermentation
(1) The fermentation pile is 13 m long, 4m wide, 1.6 + -0.1 m high and about 60m long 3 Fermentation volume, the weight of the mixed materials of a single pile is 38-40 tons;
(2) 2, an exhaust pipe is used as the fermentation air pipe, and the ventilation length of the exhaust pipe is 10 meters;
(3) drawing a boundary in a fermentation area by using paint, confirming a pile position distance, and setting the length of a real fermentation pile in a paint line;
(4) the power requirement of the control system is three-phase 380V, and the maximum power of a single station is 3.7KW.
Fermentation material preparation
1) Mixing materials by using a turner in a small dry sewage plant, sampling and detecting the mixed materials, and controlling the initial fermentation water content to be 60 +/-1;
2) The mixed material was seen to be in a fluffy state with no clumping.
3) During mixing, various auxiliary materials, microbial inoculum and nutritional agents are added into a single pile.
Fermentation and piling process
1) The stacking is carried out by using a loader, and the loader is not suitable for slow dumping when dumping materials.
2) And (4) pouring materials along a paint line after building the pile, and after basically forming, manually repairing the pile to ensure the rule of the pile body.
Fermentation process control
1) The air flow film is not covered (the sand filling PE pipe and the sand bag are used for edge pressing) for 1-2 days during pile building, the temperature of the pile body is observed and recorded 4-6 times a day, and the air flow film is covered after the temperature of the whole pile is raised and is relatively uniform.
2) The temperature is observed at least more than 4 times every day, the air quantity is automatically adjusted according to the temperature condition, and proper intervention is manually carried out.
The fermentation is finished
And (4) predicting that the single pile is fermented for about 30 days, opening the airflow membrane to check the pile condition, and if the fermentation basically meets the requirement, ending the fermentation and comprehensively sampling the materials for detection. The fermentation maturity criteria are shown in tables 3-2:
TABLE 3-2 fermentation maturity criteria
3.2 fermentation Process index detection Process and method
The sludge and mussel shell combined film covering aerobic composting experiment has a period of 26 days, four points are randomly taken from a pile body respectively at 0,1,3,6,9, 12, 17, 22, 25 and 28 hours, 0.5 kg of sample is taken at a position 20-30cm away from each point, the samples are uniformly mixed, then 0.5 kg of sample is taken, the mixture is refrigerated at 4 ℃, and 11 samples are collectively transported to a smoke table for determination.
The sample is divided into two parts, one part is used for measuring physicochemical properties such as pH, conductivity (EC), water content (MC), volatile Solid (VS), water soluble organic carbon (WSC), ammonium Nitrogen (NH) 4+ -N), nitrate Nitrogen (NO) 3 -N) and Germination Index (GI), etc. The other part of the sample was freeze-dried, ground to a powder, stored at-20 ℃ and used to measure cellulose, lignin content, microbial biomass (-80 ℃), kjeldahl nitrogen and Total Phosphorus (TP) total potassium (TK). The specific measurement method is as follows:
(1) Water content and organic matter
Water content: placing 30mL of clean crucible in a 105 ℃ oven for 0.5 hour, cooling to room temperature, and repeatedly weighing until constant weight is recorded as m 1 (g) Taking a proper amount of fresh compost sample to a crucible, and recording the weight of the sample and the crucible as m 2 (g) Drying in a 105 deg.C oven for 24 hr, cooling to room temperature, weighing and recording as m 3 (g) Continuing the oven drying for 24 hours, and marking the weighing mark as m 3 Several times until Δ mx-1<5% Δ mx-2, that is to say that the change in the weighed mass of the day is less than 5% of the change in the mass of the previous day. The water content of the compost is calculated according to the following formula:
MC(％)＝(m 2 -m 3 )/(m 2 -m 1 )×100％
organic matter: the organic matter of the experiment is expressed by VS content, the dried sample is placed in a muffle furnace, temperature is programmed to 550 ℃, ignition is carried out for 6 hours, and the sample is weighed and recorded as m after being cooled to room temperature 4 (g) Then, the calculation formula of VS and the degradation efficiency is as follows:
VS(％)＝(m 3 -m 4 )/(m 2 -m 1 )×100％
VS degradation efficiency＝(VS i -VS t )/VS i ×100％
wherein VS is i (g) The amount of organic matter at the beginning of composting, VS t (g) The amount of organic matter is the amount of organic matter over time t.
(2)pH
Weighing 3.00g of a fresh compost sample, adding the fresh compost sample into a 50mL conical flask, adding 30mL of distilled water according to a solid-liquid ratio of 1.
(3) Water soluble carbon and water soluble nitrogen
Centrifuging the compost extract at 10000rpm for 10min, filtering the supernatant with a 0.45-micrometer needle filter, diluting with distilled water by a proper multiple, and measuring the content of water-soluble carbon in the sample with a TOC analyzer; with ammonia nitrogen kit (LH-N) 2 /N 3 -100, lian-huaTech.Co., ltd.) colorimetry and ultraviolet spectrophotometer can be used for measuring the water-soluble ammonium nitrogen and the water-soluble nitrate nitrogen content ammonium nitrogen and nitrate nitrogen by ion chromatography.
(4) Index of germination
Sucking 2mL of the filtrate, injecting the filtrate into a disposable culture dish with filter paper spread at the bottom, uniformly placing 10 full-grain pakchoi seeds with approximate size on each piece of filter paper, performing a seed germination experiment by taking equivalent distilled water as a control, and repeating the steps for two times in each group. Placing the culture dish in a constant temperature box at 20 ℃ for culturing for 48 hours in a dark place, and counting the germination rate and the root length of the seeds, wherein the calculation formula of the germination rate and the germination index is as follows:
in the formula:
GI: germination index,%;
GR: germination percentage of seeds cultured by the sample leaching liquor;
l: the average root length of the seeds cultured by the sample leaching liquor is cm;
GR 0 : the germination percentage of the seeds cultured in distilled water is percent;
L 0 : average root length, cm, of seeds cultured in distilled water.
The results are retained to integers.
(5) Index of smell
A sensory evaluation method is adopted, and according to a preset aerobic fermentation odor evaluation standard, more than 3 odor evaluation groups evaluate and score the odor condition of a fermentation site at positions 1 meter, 10 meters, 50 meters, 100 meters and 200 meters away from the fermentation site, wherein the odor evaluation standard is shown in the following table.
Tables 3-3 odor evaluation criteria
3.3 Pilot plant test results and analysis of aerobic fermentation
(1) Odor change during aerobic fermentation
As shown in FIG. 2, only in the early stage of fermentation, the odor is more obvious within 1 meter from the stack, and no unpleasant odor is smelled in the distance of more than 10 meters or in the middle and later stages of fermentation. Therefore, the film-covering fermentation process has better control on smell in the fermentation process.
(2) Temperature change in aerobic fermentation process
As shown in figure 3, the fermentation process is started quickly due to the addition of the special leaven which is suitable for the characteristics of the sludge and the shells, the fermentation process starts to enter the high-temperature fermentation process from 3 days, the high-temperature fermentation process is basically maintained at more than 50 ℃, and the high-temperature period is maintained for more than 10 days.
(3) Aerobic fermentation water content and organic matter change
As shown in FIG. 4, the water content and organic matter index of the material continuously decreased at a faster rate during the fermentation process, indicating that the fermentation process was better controlled.
(4) Aerobic fermentation pH and C/N ratio variation
As shown in FIG. 5, the pH decreased and then gradually increased slightly during composting due to the decrease in organic acids produced by the decomposition of macromolecular organic materials during the initial stage of fermentation. According to the determination that the mussel shell powder is an alkaline material, which may be a reason for the gradual small increase of pH in the middle and later stages of fermentation, in addition, organic nitrogen in the fermentation process is continuously converted into ammonium nitrogen, and the ammonium nitrogen is also converted into nitrate nitrogen to form a dynamic balance, so that the gradual small increase of pH is caused.
As shown in FIG. 6, the carbon-nitrogen ratio in the fermentation process is in a gradually decreasing trend, which accords with the general rule of aerobic composting and shows that carbon and nitrogen are being utilized by fermenting microorganisms according to a certain ratio, and the decrease of the carbon-nitrogen ratio indicates that the sludge fermentation tends to mature.
(5) Nutrient change in aerobic fermentation
As shown in fig. 7, the total nutrient (total nutrient means the sum of nitrogen, phosphorus and potassium nutrients in the fermentation material) content in the fermentation process is increased by a small amount as a whole, which is caused by that part of organic materials are decomposed and utilized and the water content of the materials is reduced to relatively increase inorganic nutrients such as nitrogen, phosphorus and potassium.
(6) Wheat germination rate experiment of aerobic fermentation product
Taking sludge and mussel shells on the 26 th day, and covering with a film, and performing aerobic fermentation and decomposition to obtain fertilizers, wherein the concentration of each group of experimental groups is 0.05mg/mL; after standing overnight, the supernatant was collected by centrifugation, and the supernatant was aspirated into a petri dish loaded with wheat malt and observed for three days, the results are shown in FIG. 8. The result finally shows that the germination rate and germination index indexes of the sludge and mussel shell combined film-covered aerobic fermentation decomposed fertilizer completely meet the safety requirements of agricultural application.
(7) Physicochemical and heavy metal indexes of aerobic fermentation
And (4) carrying out heavy metal index detection on the aerobic fermentation sample (fermented to 26 days). The lower graph shows the contrast between the test data and each standard. The indexes show that harmful indexes such as heavy metal completely meet the index requirements of products such as organic fertilizers, but indexes of nitrogen, phosphorus and potassium and organic matters are slightly lower than the index requirements of the organic fertilizers, because the main component of the shell powder is calcium carbonate, organic matters are basically not contained, and the organic matter content of the whole fermentation product is reduced, so that the shell powder and sludge combined fermentation product needs to be sold as the organic fertilizer, a subsequent aging process is needed, and nitrogen, phosphorus and potassium and a small part of high organic matter material need to be properly supplemented.
Tables 3-4 physicochemical indices and heavy Metal comparison
Example 4
4.1 list of raw materials see Table 4-1:
TABLE 4-1 list of aerobic fermentation raw materials of shell powder
4.2 Experimental results and analysis of aerobic fermentation pilot scale (the analysis method is the same as that in example 3), and the period of the experiment of covering membrane aerobic composting by combining the sludge and the mussel shell is 26 days.
(1) Odor change during aerobic fermentation
Only in the initial fermentation stage, the odor is obvious within 1 meter of the stack body, and no unpleasant odor is smelled in the middle and later fermentation stages or at a distance of more than 10 meters. Therefore, the film-covering fermentation process has better control on smell in the fermentation process.
(2) Temperature change during aerobic fermentation
The special leaven adapted to the characteristics of the sludge and the shells is added, so that the fermentation process is started quickly, the fermentation process starts to enter a high-temperature fermentation process on the 2 nd day from the beginning of fermentation, the high-temperature fermentation process is basically maintained at more than 53 ℃, and the high-temperature period is maintained for more than 10 days.
(3) Aerobic fermentation water content and organic matter change
The water content and organic matter indexes of the materials in the fermentation process continuously decrease at a higher speed, which indicates that the fermentation process is better controlled.
(4) Aerobic fermentation pH and C/N ratio variation
In the composting process, the pH value is reduced and then gradually and slightly increased, which is caused by the reduction of organic acid generated by the decomposition of macromolecular organic materials in the initial fermentation stage.
The carbon-nitrogen ratio in the fermentation process is in the trend of gradual decrease, and the general rule of aerobic composting is met.
(5) Nutrient change of aerobic fermentation
The total nutrient (the total nutrient refers to the sum of nitrogen, phosphorus and potassium nutrients in the fermentation material) content in the fermentation process is improved to a small extent on the whole, because part of organic materials are decomposed and utilized and the water content of the materials is reduced, so that the inorganic nutrients such as nitrogen, phosphorus and potassium are relatively increased.
(6) Wheat germination rate experiment of aerobic fermentation product
Taking a sample on the 27 th day, and configuring the concentration of each experimental group to be 0.04mg/mL; standing overnight, centrifuging to obtain supernatant, sucking the supernatant, placing into a culture dish with wheat malt, and observing for three days until the final germination rate is 72.5% and the germination index is 91.3%; the result shows that the germination rate and germination index indexes of the sludge and mussel shell combined film-coated aerobic fermentation decomposed fertilizer finally completely meet the safety requirement of agricultural application.
(7) Aerobic fermentation physicochemical and heavy metal indexes of shell powder
And (4) carrying out heavy metal index detection on the shell powder aerobic fermentation sample (fermented to 26 days). Harmful indexes such as heavy metal and the like completely meet the index requirements of products such as organic fertilizers and the like.
Example 5
5.1 List of materials see Table 5-1:
TABLE 5-1 list of aerobic fermentation raw materials of shell powder
5.2 aerobic fermentation pilot-scale experiment result and analysis (the analysis method is the same as that in example 3), the sludge and mussel shells are combined to be coated with a film for aerobic composting experiment, and the period is 25 days.
(1) Odor change during aerobic fermentation
Only in the initial fermentation stage, the odor is obvious within 1 meter of the stack body, and no unpleasant odor is smelled when the distance is more than 10 meters or in the middle and later fermentation stages. Therefore, the film-covering fermentation process has better control on the smell in the fermentation process.
(2) Temperature change during aerobic fermentation
The special leaven adapting to the characteristics of the sludge and the shells is added, so that the fermentation process is started quickly, the fermentation starts to enter the high-temperature fermentation process 2 days after the fermentation starts, the high-temperature fermentation process is basically maintained at more than 55 ℃, and the high-temperature period is maintained for more than 10 days.
(3) Aerobic fermentation water content and organic matter change
The water content and organic matter indexes of the materials in the fermentation process continuously decrease at a higher speed, which indicates that the fermentation process is better controlled.
(4) Aerobic fermentation pH and C/N ratio variation
The pH value is reduced and then gradually increased slightly in the composting process, which is caused by the reduction of organic acid generated by the decomposition of macromolecular organic materials in the early fermentation stage.
The carbon-nitrogen ratio in the fermentation process is in a gradual descending trend, and the general rule of aerobic composting is met.
(5) Nutrient change in aerobic fermentation
The total nutrient (the total nutrient refers to the sum of nitrogen, phosphorus and potassium nutrients in the fermentation material) content in the fermentation process is improved to a small extent on the whole, because part of organic materials are decomposed and utilized and the water content of the materials is reduced, so that the inorganic nutrients such as nitrogen, phosphorus and potassium are relatively increased.
(6) Wheat germination rate experiment of aerobic fermentation product
Taking a sample on the 25 th day, and configuring the concentration of an experimental group to be 0.04mg/mL; standing overnight, centrifuging to obtain supernatant, sucking the supernatant, placing into a culture dish with wheat malt, and observing for three days to obtain a final germination percentage of 83.9% and a germination index of 111.7%; the result shows that the germination rate and germination index indexes of the sludge and mussel shell combined film-covered aerobic fermentation decomposed fertilizer finally completely meet the safety requirement of agricultural application.
(7) Physicochemical and heavy metal indexes of aerobic fermentation
And (4) carrying out heavy metal index detection on the shell powder aerobic fermentation sample (fermented to the 25 th day). Harmful indexes such as heavy metal and the like completely meet the index requirements of products such as organic fertilizer and the like.
Example 6
6.1 List of raw materials see Table 6-1:
TABLE 6-1 List of aerobic fermentation raw materials of shell powder
6.2 Experimental results and analysis of aerobic fermentation pilot scale (the analysis method is the same as that in example 3), and the period of the experiment of covering membrane aerobic composting by combining the sludge and the mussel shell is 24 days.
(1) Odor change during aerobic fermentation
Only in the initial fermentation stage, the odor is obvious within 1 meter of the stack body, and no unpleasant odor is smelled in the middle and later fermentation stages or at a distance of more than 10 meters. Therefore, the film-covering fermentation process has better control on the smell in the fermentation process.
(2) Temperature change in aerobic fermentation process
Due to the addition of the special decomposing leaven which is suitable for the characteristics of the sludge and the mussel shells, the fermentation process is started quickly, the fermentation process starts to enter the high-temperature fermentation process at the 2 nd day, the high-temperature fermentation process is basically maintained at more than 57 ℃, and the high-temperature period is maintained for more than 10 days.
(3) Aerobic fermentation water content and organic matter change
The water content and organic matter indexes of the materials in the fermentation process continuously decrease at a higher speed, which indicates that the fermentation process is better controlled.
(4) Aerobic fermentation pH and C/N ratio variation
The pH value is reduced and then gradually increased slightly in the composting process, which is caused by the reduction of organic acid generated by the decomposition of macromolecular organic materials in the early fermentation stage.
The carbon-nitrogen ratio in the fermentation process is in the trend of gradual decrease, and the general rule of aerobic composting is met.
(5) Nutrient change in aerobic fermentation
The total nutrient (the total nutrient refers to the sum of nitrogen, phosphorus and potassium nutrients in the fermentation material) content in the fermentation process is improved to a small extent on the whole, because part of organic materials are decomposed and utilized and the water content of the materials is reduced, so that the inorganic nutrients such as nitrogen, phosphorus and potassium are relatively increased.
(6) Wheat germination rate experiment of aerobic fermentation product
Taking a sample on the 24 th day, and configuring the concentration of each group of experimental groups to be 0.04mg/mL; standing overnight, centrifuging to obtain supernatant, sucking the supernatant, placing into a culture dish with wheat malt, and observing for three days until the final germination rate is 100% and the germination index is 142.3%; the result shows that the germination rate and germination index indexes of the sludge and mussel shell combined film-coated aerobic fermentation decomposed fertilizer finally completely meet the safety requirement of agricultural application.
(7) Physicochemical and heavy metal indexes of aerobic fermentation
And (4) carrying out heavy metal index detection on the sludge and mussel shell combined film-coated aerobic fermentation decomposed fertilizer (fermented to the 24 th day). Harmful indexes such as heavy metal and the like completely meet the index requirements of products such as organic fertilizer and the like.
While embodiments of the invention have been described above, it is not intended to be limited to the details shown, described and illustrated herein, but is to be accorded the widest scope consistent with the principles and novel features herein disclosed, and to such extent that such modifications are readily available to those skilled in the art, and it is not intended to be limited to the details shown and described herein without departing from the general concept as defined by the appended claims and their equivalents.
<110> institute of environmental science of China
<120> decomposed leaven and application thereof
<160>1
<210>1
<211>1400
<212>DNA
<213> Artificial sequence
<220>
<400>1
agaagcttgc ttctcttgag agcggcggac gggtgagtaa tgcctaggaa tctgcctagt 60
ggtgggggat aacgttcgga aacggacgct aataccgcat acgtcctacg ggagaaagcg 120
ggggaccttc gggcctcgcg ccattagatg agcctaggtc ggattagcta gttggtgagg 180
taaaggctca ccaaggcgac gatccgtaac tggtctgaga ggatgatcag tcacactgga 240
actgagacac ggtccagact cctacgggag gcagcagtgg ggaatattgg acaatgggcg 300
aaagcctgat ccagccatgc cgcgtgtgtg aagaaggtct tcggattgta aagcacttta 360
agttgggagg aagggtagta acttaatacg ttgctacttt gacgttaccg acagaataag 420
caccggctaa cttcgtgcca gcagccgcgg taatacgaag ggtgcaagcg ttaatcggaa 480
ttactgggcg taaagcgcgc gtaggtggtt cagtaagttg gatgtgaaat ccccgggctc 540
aacctgggaa ctgcatccaa aactgctgaa ctagagtacg gtagagggtg gtggaatttc 600
ctgtgtagcg gtgaaatgcg tagatatagg aaggaacacc agtggcgaag gcgaccacct 660
ggactgatac tgacactgag gtgcgaaagc gtggggagca aacaggatta gataccctgg 720
tagtccacgc cgtaaacgat gtcaactagc cgttgggagt cttgaactct tagtggcgca 780
gctaacgcat taagttgacc gcctggggag tacggccgca aggttaaaac tcaaatgaat 840
tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc 900
ttacctggcc ttgacatgct gagaactttc tagagataga ttggtgcctt cgggaactca 960
gacacaggtg ctgcatggct gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgt 1020
aacgagcgca acccttgtcc ttagttacca gcacgtaatg gtgggaactc taaggagact 1080
gccggtgaca aaccggagga aggtggggat gacgtcaagt catcatggcc cttacggcca 1140
gggctacaca cgtgctacaa tggtcggtac aaagggttgc caagccgcga ggtggagcta 1200
atcccataaa accgatcgta gtccggatcg cagtctgcaa ctcgactgcg tgaagtcgga 1260
atcgctagta atcgtgaatc agaatgtcac ggtgaatacg ttcccgggcc ttgtacacac 1320
cgcccgtcac accatgggag tgggttgcac cagaagtagc tagtctaacc gcaaggggga 1380
cggttaccac ggtgtgatcg 1400
Claims (10)
1. Pseudomonas sp LZ4-5, characterized in that it has the deposit number:
CGMCC NO.21761。
2. a decomposed leaven comprising the Pseudomonas sp as set forth in claim 1
LZ4-5。
3. The decomposing inoculant according to claim 2, further comprising: at least one of bacillus subtilis and yeast.
4. The decomposing starter as claimed in claim 3, wherein the decomposing starter comprises 1 part of bacillus subtilis, 1 part of yeast and 2 parts of Pseudomonas (Pseudomonas sp.) LZ4-5 by weight.
5. Use of Pseudomonas (Pseudomonas sp.) LZ4-5 according to claim 1 in preparation of sludge and mussel shell combined film-covered aerobic fermentation decomposed fertilizer.
6. The use of the decomposed starter culture according to any one of claims 2 to 4 in the preparation of a sludge and mussel shell combined film-coated aerobic fermentation decomposed fertilizer.
7. The use according to claim 6, wherein the starter culture is present in an amount of 0.3% to 0.4% by weight of the total fermentation feedstock.
8. The use of claim 7, wherein the total fermentation feedstock further comprises, in weight percent:
38-40% of sludge, 13-15% of rice hull powder and 12-14% of mushroom dregs; 30 to 36 percent of shell powder and 0.6 to 0.7 percent of urea.
9. The use according to claim 8, wherein the sludge has a water content of 80-85%, the rice hull powder has a water content of 15-20%, the mushroom dregs have a water content of 35-40%, the shell powder comprises a fine shell powder having a particle size of 40-60 mesh and a coarse shell powder having a particle size of 2-5 mesh, and the weight ratio of the fine shell powder to the coarse shell powder is 5.
10. A sludge prepared by applying the decomposed leaven of any one of claims 2 to 4 and mussel shells to be combined with a film for aerobic fermentation of decomposed fertilizer.
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| CN102559544A (en) * | 2011-12-14 | 2012-07-11 | 青岛思普润水处理有限公司 | Pseudomonas sp. for removing ammonia nitrogen in sewage under low-temperature condition and isolated culture method |
| CN108101320A (en) * | 2017-12-18 | 2018-06-01 | 佛山市铁人环保科技有限公司 | A kind of Sludge deodorant and preparation method thereof, deodorizing method |
| CN110316751A (en) * | 2019-07-03 | 2019-10-11 | 中国建筑材料科学研究总院有限公司 | A kind of conch meal and preparation method thereof |
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