CN113549568A - Cell-lysing strain, microbial inoculum for sludge reduction and application thereof - Google Patents

Abstract

The invention relates to a cell dissolving strain, a microbial inoculum for sludge reduction and application thereof, wherein the cell dissolving strain is bacillus terrae (Bacillus (A) (B))Geobacillus sp.) THE THE-14 with THE preservation number of CCTCC M2021514 is separated and screened from sludge matured by high-temperature acclimation, THE cell lysing strain and a microbial inoculum containing THE cell lysing strain can be applied to THE reduction treatment of THE sludge, and THE cell lysing strain separated and screened by THE invention is Geobacillus (Bacillus licheniformis) ((R))Geobacillus sp.) THE THE-14 can utilize organic matters in THE sludge to grow and reproduce, and THE enzyme generated in THE metabolic process of THE strain can accelerate THE hydrolysis of extracellular polymeric substances, thereby achieving THE purpose of sludge cell lysis.

Description

Cell-lysing strain, microbial inoculum for sludge reduction and application thereof

Technical Field

The invention belongs to the field of sludge biological treatment, and particularly relates to a cell-lysing strain, a microbial inoculum for sludge reduction and application thereof.

Background

Biological wastewater treatment plants (WWTP) have been used worldwide to treat municipal wastewater. Although effective in removing organic matter from wastewater, a large amount of excess sludge is produced. With the increasing popularity and population of sewage treatment plants, the construction of new sewage treatment plants and the stricter sewage treatment requirements, the production of sludge will continue to increase. Thus, the treatment and disposal of sludge has become an increasingly significant challenge in the field of environmental engineering.

In a conventional sewage treatment plant, there are mainly two methods of reducing the amount of remaining sludge, i.e., a wastewater line and a sludge line. The method is characterized in that return sludge after cell lysis is recycled to a mainstream bioreactor for further biodegradation, and physical, chemical and biological means are utilized to minimize the sludge discharged outwards by the whole sewage treatment system, so that the method has good application prospect. Physical and chemical methods result in additional energy consumption, high costs, secondary pollution and other economic costs. In contrast, biological pretreatment shows absolute advantages in accelerating the sludge hydrolysis process.

Extracellular Polymeric Substance (EPS) is an important component of a sludge floc matrix and is composed of various organic substances, such as organic macromolecules, such as polysaccharide, protein, humic acid, uronic acid, lipid compounds and the like. The sludge reduction by the biological pretreatment method is realized by adding hydrolase into the sludge, accelerating the hydrolysis of extracellular polymers under the catalysis of the hydrolase, destroying the cell structure of microorganisms and dissolving insoluble substances in the sludge. However, the direct addition of the hydrolase has the problems of high cost, harsh reaction conditions and high difficulty in large-scale practical application. Therefore, in order to increase the industrial applicability of the biological pretreatment method, it is necessary to isolate and screen a strain capable of reducing the sludge, and the hydrolysis of extracellular polymers is accelerated by enzymes produced by the strain in the metabolic process of the sludge, thereby achieving the purpose of reducing the sludge.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide a cell dissolving strain, a bacterial agent for sludge reduction and application thereof.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

the first technical scheme is as follows:

a cytolytic strain is Geobacillus sp (THE-14), belongs to Geobacillus sp (Geobacillus sp), and is preserved in China center for type culture Collection with THE preservation address: wuhan university in Wuhan, China, with a preservation time of 2021 year, 5 months and 11 days; the preservation number is CCTCC M2021514.

Further, THE 16SrDNA sequence of THE Geobacillus sp (Geobacillus sp.) THE-5 is shown as SEQ ID: 1 is shown.

Further, the lysate strain is obtained by separating and screening sludge matured by high-temperature acclimatization.

Further, the sludge domesticated at high temperature is obtained by domesticating sewage sludge at 55-80 ℃ under the aeration condition.

Furthermore, in the acclimatization process, the aeration rate is 0.1-1.0vvm, and the acclimatization time is 5-60 days.

The second technical scheme is as follows:

a microbial inoculum for sludge reduction comprising said lysate strain.

Further, THE microbial inoculum for sludge reduction also comprises a bacillus terreus (Geobacillus sp.) THE-4 strain and/or a bacillus terreus (Geobacillus sp.) THE-5 strain;

THE Geobacillus sp (Geobacillus sp.) THE-4 belongs to Geobacillus sp (Geobacillus sp.) and is preserved in China center for type culture Collection with THE preservation address: wuhan university in Wuhan, China, with a preservation time of 2021 year, 5 months and 11 days; the preservation number is CCTCC M2021512;

THE Geobacillus sp (Geobacillus sp.) THE-5 belongs to Geobacillus sp (Geobacillus sp.) and is preserved in China center for type culture Collection with THE preservation address: wuhan university in Wuhan, China, with a preservation time of 2021 year, 5 months and 11 days; the preservation number is CCTCC M2021513.

The third technical scheme is as follows:

an application of the microbial inoculum for sludge reduction in sludge reduction treatment.

The technical scheme is as follows:

a method for sludge reduction treatment by using the microbial inoculum for sludge reduction comprises the following steps:

step one, activated culture of the lysate:

inoculating the lysate strain to a liquid medium, and performing constant temperature shaking culture for activation to obtain OD₆₀₀Then the seed liquid is added into the seed liquid of 1.0-1.2, and then the centrifugal separation is carried out, and the thalli are collected.

Step two, uniformly blowing and beating the collected thalli by using sterile water to obtain a bacterial liquid;

step three, sludge reduction treatment

Adding bacterial liquid into the organic sludge to be treated, adjusting the pH value to 6-10, and performing lysis reaction under aeration and stirring conditions.

Further, in step one, the inoculum size of the lysate in said liquid medium is 1-5%; the temperature of the shaking culture is 50-85 ℃, the shaking speed is 40-200rpm, the times of the constant temperature shaking culture are 2-3 times, and the time of each shaking culture is 6-20 h;

in the second step, the OD of the bacterial liquid₆₀₀The value is between 0.4 and 0.8,

in the third step, the adding amount of the bacterial liquid is 10-15% of the volume of the organic sludge to be treated, the temperature of the lysis reaction is 50-85 ℃, the aeration rate is 0.1-1.0vvm, the stirring rate is 50-200rpm, and the time of the lysis reaction is 1-36 h.

Compared with the prior art, the invention has the following beneficial effects:

1. THE invention adopts THE sludge domesticated and matured at THE temperature of 55-80 ℃ to separate and screen THE lysis bacterial strain, THE lysis bacterial strain Geobacillus sp THE-14 separated by THE method can invisibly grow in THE sludge to be treated, and secrete extracellular enzymes, thereby achieving THE purpose of promoting THE hydrolysis of macromolecular organic matters in THE sludge, and releasing organic matters in a solid phase of THE sludge into a liquid phase for subsequent utilization.

2. When the microbial inoculum is used for lysing organic sludge, the lysis effect of the sludge is greatly improved by adding 2-valent metal ions.

Compared with the traditional physical and chemical reduction technology, the cell-dissolving strain and the sludge treatment method provided by the invention have the characteristics of economy, high efficiency, low energy consumption, high cell-dissolving efficiency and easily controlled reaction conditions.

Drawings

FIG. 1 is a phylogenetic tree of cytolytic strains;

FIG. 2 is a graph showing changes in protease activity of each strain in example 2;

FIG. 3 is a graph comparing the degradation rate of VSS of sludge by each strain in example 3;

FIG. 4 is a graph showing the effect of temperature on the degradation rate of sludge VSS in example 4.

Detailed Description

The invention is further illustrated by the following examples.

THE lysis bacterial strain THE-14 of THE invention is separated and screened by sewage sludge which is derived from: second-phase return sludge pump house of bridge north of Nanjing City (A)²the/O process); indexes of sewage sludge: the TSS is 13285 +/-146 mg/L; 6003 +/-30 mg/L of VSS; TCOD is 4350 +/-88 mg/L; SCOD is 65 plus or minus 5 mg/L; the pH value is 6.8 plus or minus 0.2;

THE lysis bacterial strain THE-4 of THE invention is separated and screened by sewage sludge which is derived from: nanjing, Qiaobei processing plant A²the/O process reflows the sludge in the sludge pump room; indexes of sewage sludge: the TSS is 13285 +/-146 mg/L; 6003 +/-30 mg/L of VSS; TCOD is 4350 +/-88 mg/L; SCOD is 65 plus or minus 5 mg/L; the pH value is 6.8 plus or minus 0.2;

THE lysis bacterial strain THE-5 of THE invention is separated and screened by sewage sludge which is derived from: a process of a Jiangxin continent sewage treatment plant (A/O) in Nanjing; indexes of sewage sludge: 10147 +/-210 mg/L TSS; VSS 7416 + -170 mg/L; TCOD 5476 + -180 mg/L; SCOD is 75 +/-10 mg/L; the pH value is 6.8 +/-0.2.

The sewage sludge used in the lysis test of the invention comes from: a. the²the/O process operates residual sludge from sewage plants, the properties of which are as follows: total Suspended Solids (TSS) is 14685 + -146 mg/L, Volatile Suspended Solids (VSS) is 6021 + -25 mg/L, Total Chemical Oxygen Demand (TCOD) is 4350 + -88 mg/L, dissolved chemical oxygen demand is 65 + -5 mg/L, and pH is 7.0-7.3. Sieving the sludge to remove particles with the particle size of more than 1mm, and then storing in a refrigerator at 4 ℃, wherein the storage period is not more than 1 week.

The reference strain YL-1 is preserved in China center for type culture Collection with the preservation number: CCTCC AB 2021109;

liquid culture medium: macroelements: nitrilotriacetic acid 100mg, NaCl 8mg, KH₂PO₄ 111mg， MgSO₄·7H₂O100 mg, peptone 1g, yeast extract 1 g; trace elements: na (Na)₂MoO₄·2H₂O 0.025 mg，FeCl₃ 0.28mg，CuSO₄ 0.16mg，MnSO₄·H₂O 2.2mg，H₃BO₃ 0.5mg， ZnSO₄·7H₂O 0.5mg，CoCl₂·6H₂O 0.046mg，CaSO₄60 mg. Adding 1L deionized water, adjusting pH to 7-7.5, placing in autoclave, and sterilizing at 121 deg.C for 20 min.

Example 1: separation, screening, identification and application of dominant cytolytic bacteria

The embodiment mainly comprises the following parts, namely, separating and screening the domesticated sludge to obtain dominant strains, and identifying the dominant strains; determining the species relationship of the dominant strains; and thirdly, obtaining the optimal process for carrying out sludge reduction treatment by utilizing the screened dominant bacterial strain through process optimization.

Firstly, separating and screening the domesticated sludge to obtain the dominant bacterial strain, and specifically comprises the following steps:

step 1, sludge domestication: acclimating the sewage sludge at the aeration condition of 0.1-1.0vvm and the temperature of 55-80 ℃ for 5-60 days;

step 2, after the domesticated mature sludge is subjected to gradient dilution, a dilution pouring method and a three-zone scribing method are adopted for separation and purification, a strain of a single colony is obtained, and the strain is named and numbered respectively;

step 2.1, pouring the plate: solid medium: macroelements: nitrilotriacetic acid 100mg, NaCl 8mg, KH₂PO₄111mg，MgSO₄·7H₂O100 mg, peptone 1g, yeast extract 1 g; 20g of agar; trace elements: na (Na)₂MoO₄·2H₂O 0.025mg，FeCl₃ 0.28mg，CuSO₄ 0.16mg， MnSO₄·H₂O 2.2mg，H₃BO₃ 0.5mg，ZnSO₄·7H₂O 0.5mg，CoCl₂·6H₂O 0.046mg， CaSO₄60 mg. Adding 1L deionized water, adjusting pH to 7-7.5, placing in autoclave, and sterilizing at 121 deg.C for 20 min. And when the temperature is cooled to 50-55 ℃, pouring the mixture into a sterile culture dish in sequence, ensuring that the mixture is fully paved at about 2/3 of the dish bottom, and inversely buckling the flat plate on a sterile table when agar is solidified.

Step 2.2, separation: taking a plurality of sterilized EP tubes, respectively adding 0.9ml of sterile water, taking 0.1ml of sludge domesticated and matured at 65 ℃, adding the sludge into a first tube of sterile water, fully mixing and shaking up, taking 0.1ml of diluent from the first tube to the next sterilized EP tube, mixing and shaking up, so diluting to a fourth tube and a fifth tube, wherein the dilution times are respectively as follows from the first tube: 10^-1、10^-2、10^-3、10^-4、10^-5(ii) a Dropwise addition 10^-4And 10^-50.1ml of each of the two tubes of diluent was placed on the corresponding plate and the diluent was coated uniformly with a sterilized coating bar. Placing the coated flat plate on a sterile table for 20-25 min, and pouring the flat plate into an incubator for 12h when the bacterial liquid permeates into the culture medium, wherein the culture temperature is 50-85 ℃. Selecting a single colony by using the sterilized inoculating loop, scribing on a flat plate by adopting a three-region scribing method, repeating for 3-4 times to obtain a purified colony, and totally separating 15 strains of the strain in the embodiment;

step 3, primarily screening the separated single bacterial colony strains through a hydrolysis ring test, retaining the bacterial strains with larger hydrolysis rings, further screening by adopting a sludge lysis test and an extracellular enzyme activity test, and retaining the bacterial strains with higher sludge VSS degradation rate and higher extracellular enzyme activity, namely the dominant bacterial strains; THE dominant strains obtained by screening are THE-14, THE-4 and THE-5;

hydrolysis ring test: inoculating the cell-dissolving strain separated from the domesticated mature sludge into a skim milk powder solid culture medium, placing the cell-dissolving strain in a constant temperature incubator, and observing the change of a hydrolysis ring after 12 hours of each strain.

The solid culture medium of the skimmed milk powder comprises the following components: macroelements: nitrilotriacetic acid 100mg, NaCl 8mg, KH₂PO₄111mg,MgSO₄·7H₂100mg of O, 1g of peptone, 1g of yeast extract, 20g of agar, 20g of skimmed milk powder and 1L of distilled water; trace elements: na (Na)₂MoO₄·2H₂O 0.025mg，FeCl₃ 0.28mg， CuSO₄ 0.16mg，MnSO₄·H₂O 2.2mg，H₃BO₃0.5mg，ZnSO₄·7H₂O 0.5mg， CoCl₂·6H₂O 0.046mg，CaSO₄60 mg. Adjusting the pH value to 7.0-7.5.

Preliminary dissolution test of sludge: taking a plurality of conical flasks, and respectively adding sludge with a solid content of 1.2-1.5%; respectively culturing the primarily screened strains for 12-15h, uniformly blowing the strains with sterile water, respectively inoculating the strains into corresponding conical flasks, carrying out shaking table culture at a constant temperature of 50-85 ℃ and an oscillation speed of 100rpm, simultaneously taking ultrapure water as a blank control, measuring the content of Volatile Suspended Solids (VSS) of sludge in each conical flask after 24h, and calculating the VSS reduction rate of the sludge.

Extracellular enzyme activity assay: and respectively inoculating the strains after primary screening into a liquid culture medium according to the inoculation amount of 1%, and determining the activity of the protease by adopting an azocasein method.

Secondly, identifying the dominant strains and determining the species relationship of the dominant strains; the method specifically comprises physiological and biochemical identification and 16S rDNA identification:

1) gram staining microscopy: gram staining microscopic examination is carried out on the dominant strain, then oxalic acid crystal violet staining solution dip dyeing is carried out for 1min, iodine solution dip dyeing is carried out for 1min, 95% ethanol decoloration is carried out for 30s, safranine solution dip dyeing is carried out for 2min-3min, a cover glass is covered, the observation is carried out under a microscope, and the observation result is that:

THE reaction conditions of THE THE-14: the thallus is dark purple and is gram positive.

THE content of THE-4: the cells were red and gram-negative.

THE reaction conditions of THE THE-5: the thallus is dark purple and is gram positive.

2)16S rDNA identification: THE base sequences of THE 16S rDNA of THE dominant strains THE-14, THE-4 and THE-5 are respectively shown in SEQ ID: 01-03, respectively.

Through identification, THE dominant strains THE-14, THE-4 and THE-5 belong to Geobacillus, and a phylogenetic tree constructed by adopting an adjacent method is shown in figure 1.

And thirdly, obtaining the optimal process for carrying out sludge reduction treatment by utilizing the screened dominant bacterial strain through process optimization.

In this example, the optimal process for sludge reduction treatment using dominant strains:

step 1, activating and culturing dominant strains:

inoculating THE dominant strain THE-14 and/or THE-4 and/or THE-5 into a liquid culture medium according to THE inoculation amount of 1-5%, and simultaneously adding sterilized sludge into THE liquid culture medium according to THE inoculation amount of 2-5%; then performing shake culture at 50-85 deg.C for 2-3 times for activation to obtain OD₆₀₀Centrifuging in 1.0-1.2 seed liquid, and collecting thallus; the time of each shaking culture is 12-15h, and the shaking speed is 100-120 rpm.

Step 2, uniformly blowing and beating the collected thalli with sterile water to obtain a bacterial liquid for later use; OD of the bacterial liquid₆₀₀0.7-0.8;

step 3, sludge reduction treatment

Adding bacterial liquid into the organic sludge to be treated according to 10-15% of the volume of the organic sludge to be treated, adjusting the pH value to 6-10, and then, stirring at 50-85 ℃ and carrying out cell lysis reaction for 1-36h under the aeration condition; the aeration rate is 0.1vvm to 1.0vvm, and the stirring rate is 40rpm to 200 rpm.

Example 2: exo-enzyme Activity analysis of dominant Strain

The residual sludge is organic wastewater containing suspended solids with high concentration, and the main components of the residual sludge comprise small-molecule soluble organic matters (monosaccharides, amino acids and the like) and non-degradable macromolecular organic matters (mainly comprising proteins, polysaccharides and lipid compounds), wherein the proportion of carbohydrates reaches 20% of the total organic matters. It was found that during the solubilization of excess sludge, glucosidases, proteases and alpha-amylases are closely related to the degradation of extracellular polymers of the sludge, and the hydrolysis of proteins is more critical. Therefore, the activity of extracellular proteases secreted by bacteria is of crucial importance.

THE dominant strains of THE invention, namely, Geobacillus sp (THE-14), Geobacillus sp (THE-4) and Geobacillus sp (THE-5), are inoculated into corresponding liquid culture media according to THE inoculation amount of 1%, and THE activity of protease is determined by adopting an azo casein method. Taking bacterial liquid at different time points, centrifuging in a desktop centrifuge (4 ℃, 10 minutes, 14,000 Xg), taking supernatant and a substrate azocasein, fully reacting for 30 minutes at 50-85 ℃, immediately adding 5% trichloroacetic acid (TCA), fully mixing for 5 seconds by using a vortex oscillator, placing under ice blocks for balancing for 10 minutes, centrifuging, taking supernatant, dropwise adding NaOH until the concentration is 0.4M, and measuring the absorbance value under 440nm by using a UV/VIS spectrophotometer. 1 protease activity unit is defined as the number of enzymes required to hydrolyze soluble casein to 1 micromole tyrosine in a unit time under certain experimental conditions. The research results show that: THE overall enzyme activity of THE THE-4 tends to be stable, 587.5U/L is reached in 14h, THE enzyme activities of THE THE-5 and THE THE-14 reach peak values which are 1500U/L and 3215U/L respectively after 6h of culture, and THE extracellular enzyme activity of THE strain begins to be in a descending state due to THE rapid consumption of nutrient substances. THE average enzyme activities of THE THE-4, THE THE-5 and THE THE-14 at 14h are 455.5U/L, 1000U/L and 1880.5U/L respectively, and THE capability of THE THE-14 for secreting extracellular enzyme activity is strongest (see figure 2).

Example 3: exploration on sludge dissolving performance of dominant strains

Respectively inoculating a mixed strain of THE dominant strain Geobacillus sp (THE-14), Geobacillus sp (THE-4), Geobacillus sp (Geobacillus sp.) THE-5, THE-14, THE-4 and THE-5 and a control strain YL-1 into corresponding solid culture media, activating and culturing for 12h at 50-85 ℃, inoculating THE activated Geobacillus sp (THE-14), Geobacillus sp (Geobacillus sp.) THE-4, Geobacillus sp (Geobacillus sp.) THE-5 and THE mixed strain of THE three into a 250ml conical flask containing 50ml of liquid culture media by using a sterile gun head, culturing for 12h at 50-85 ℃, repeatedly activating for 3 times at 120 rpm. After the activated and matured thallus is centrifuged by a desk centrifuge (6000rpm, 10min), the thallus is collected and blown uniformly by sterile water. The volume of dissolved sludge of each group is 360ml, the volume of added bacterial liquid is 40ml, an uninoculated blank control group is additionally arranged, and 40ml of distilled water is added into the uninoculated blank control group, so that the volume of the bacterial liquid accounts for 10% of the total reaction system. Then, stirring and reacting the groups for 36 hours at 50-85 ℃, pH6.8-7.0 and under aeration conditions; the aeration rate was 0.2vvm and the stirring rate was 60. + -.10 rpm.

The VSS degradation rate in each sample was measured using the national standard method (see fig. 3). The research results show that: the degradation rate of the sludge VSS in the blank control group is 4.7%. THE degradation rates of THE THE-4, THE THE-5 and THE THE-14 to THE sludge VSS are respectively 20.2%, 24.5% and 26.4%, which are respectively increased by 15.5%, 19.8% and 22% compared with a blank control group, and are respectively increased by 12.9%, 17.2% and 19.1% compared with a control strain YL-1; three strains are combined and added, and the VSS reduction rate can reach 28.8%. Referring to the protease activity in example 2, it can be seen that the higher the extracellular enzyme protease activity secreted by the bacteria, the stronger the solubility to sludge VSS.

Example 4: effect of temperature on lysis

THE dominant bacterial strains, namely Geobacillus sp (THE-14), Geobacillus sp (THE-4), Geobacillus sp (Geobacillus sp) THE-5 and THE three bacterial strains are mixed and then cultured for 15-18h at 60 ℃, 65 ℃ and 70 ℃ respectively, and are repeatedly activated for 3 times to ensure that THE OD600 is between 1.0 and 1.2, and then THE mixture is centrifuged (6000rpm and 10min) to collect THE thalli for THE subsequent lysis experiment. The temperature of each reactor was 60 ℃, 65 ℃ and 70 ℃. The volume of dissolved sludge is 360ml, the volume of added bacterial liquid is 40ml, and the volume of the bacterial liquid accounts for 10% of the total reaction system. The degradation rate of VSS at different temperatures is shown in fig. 4. It can be seen that the lysis of the sludge by the lysate was the best at 65 ℃. THE high temperature inhibits THE lysis of THE THE-4 and THE-5 to some extent. In a reactor at 65 ℃, THE degradation rates of THE THE-4, THE THE-5 and THE THE-14 to THE sludge VSS reach 20.2 percent, 24.5 percent and 26.4 percent respectively. In a three-strain composite reaction system, the degradation rate of VSS reaches 28.8%.

The embodiments described above are only preferred embodiments of the invention and are not exhaustive of the possible implementations of the invention. Any obvious modifications to the above would be obvious to those of ordinary skill in the art, but would not bring the invention so modified beyond the spirit and scope of the present invention.

Sequence listing

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agggaatctt ccgcaatgga cgaaagtctg acggagcgac gccgcgtgag cgaagaaggt 420

cttcggatcg taaagctctg ttgttaggga agaagaagta ccgttcgaat agggcggtac 480

ggtgacggta cctaacgaga aagccccggc taactacgtg ccagcagccg cggtaatacg 540

tagggggcga gcgttgtccg gaattattgg gcgtaaagcg cgcgcaggcg gtcccttaag 600

tctgatgtga aagcccacgg ctcaaccgtg gagggtcatt ggaaactggg ggacttgagt 660

gcagaagagg agagcggaat tccacgtgta gcggtgaaat gcgtagagat gtggaggaac 720

accagtggcg aaggcggctc tctggtctgt aactgacgct gaggcgcgaa agcgtgggga 780

gcaaacagga ttagataccc tggtagtcca cgccgtaaac gatgagtgct aagtgttaga 840

ggggttaaac cctttagtgc tgtagctaac gcgttaagca ctccgcctgg ggagtacggc 900

cgcaaggctg aaactcaaag gaattgacgg gggcccgcac aagcggtgga gcatgtggtt 960

taattcgaag caacgcgaag aaccttacca ggtcttgaca tcccctgaca accctggaga 1020

cagggcgttc ccccttcggg gggacagggt gacaggtggt gcatggttgt cgtcagctcg 1080

tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac cctcgcccct agttgccagc 1140

attcagttgg gcactctagg gggactgccg gctaaaagtc ggaggaaggt ggggatgacg 1200

tcaaatcatc atgcccctta tgacctgggc tacacacgtg ctacaatggg cggtacaaag 1260

ggctgcgaac ccgcgagggg gagcgaatcc caaaaagccg ctctcagttc ggattgcagg 1320

ctgcaactcg cctgcatgaa gccggaatcg ctagtaatcg cggatcagca tgccgcggtg 1380

aatacgttcc cgggccttgt acacaccgcc cgtcacacca cgagagcttg caacacccga 1440

agtcggtgag gtaacccttt aagggagcca gccgccgaag gtggggcaag tgattggggt 1500

gaagtcgtaa caaggtaacc a 1521

Claims (10)

1. A cell-lysing strain, which is characterized by comprising Bacillus terreus (Geobacillus sp.) THE THE-14 is preserved in China center for type culture Collection with THE preservation number of CCTCC M2021514.

2. The lysate strain of claim 1, wherein the Geobacillus bacterium (C.), (C.), (C.Geobacillus sp.) THE 16S rDNA sequence of THE THE-5 is shown in SEQ ID: 1 is shown.

3. The lysate strain of claim 1, wherein the lysate strain is isolated and screened from a high temperature acclimated mature sludge.

4. The lysate strain according to claim 1, wherein the high-temperature acclimated mature sludge is obtained by acclimating sewage sludge at 55-80 ℃ under aeration conditions.

5. A lysate strain according to claim 4, characterised in that during acclimatisation, the aeration rate is between 0.1 and 1.0vvm and the acclimatisation time is between 5 and 60 days.

6. A microbial inoculum for sludge reduction comprising the lysate strain according to claim 1.

7. The microbial agent for sludge reduction according to claim 6, further comprising a Bacillus terreus strain (Geobacillus) as a lysate strainGeobacillus sp.) THE THE-4 and/or THE lytic strain Geobacillus (Geobacillus)Geobacillus sp.）THE-5；

Said Bacillus terrae: (A), (B), (C)Geobacillus sp.) THE THE-4 is preserved in China center for type culture Collection with a preservation number of CCTCC M2021512;

said Bacillus terrae: (A), (B), (C)Geobacillus sp.) THE THE-5 is preserved in China center for type culture Collection with a preservation number of CCTCC M2021513.

8. Use of the microbial agent for sludge reduction according to any one of claims 6 to 7 in sludge reduction treatment.

9. A method for sludge reduction treatment using the microbial agent for sludge reduction according to any one of claims 6 to 7, comprising the steps of:

step one, activated culture of the lysate:

inoculating the lysate strain to a liquid medium, and performing constant temperature shaking culture for activation to obtain OD₆₀₀Centrifuging in 1.0-1.2 seed liquid, and collecting thallus;

step two, uniformly blowing and beating the collected thalli by using sterile water to obtain a bacterial liquid;

step three, sludge reduction treatment

Adding bacterial liquid into the organic sludge to be treated, adjusting the pH value to 6-10, and performing lysis reaction under aeration and stirring conditions.

10. The method for sludge reduction treatment using a lysate strain according to claim 9,

in step one, the inoculum size of the lysate in said liquid medium is 1-5%; the temperature of the shaking culture is 50-85 ℃, the shaking speed is 40-200rpm, the times of the constant temperature shaking culture are 2-3 times, and the time of each shaking culture is 6-20 h;

in the second step, the OD of the bacterial liquid₆₀₀The value is between 0.4 and 0.8,

in the third step, the adding amount of the bacterial liquid is 10-15% of the volume of the organic sludge to be treated, the temperature of the lysis reaction is 50-85 ℃, the aeration rate is 0.1-1.0vvm, the stirring rate is 50-200rpm, and the time of the lysis reaction is 1-36 h.

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