CN110218682A - One plant of pseudomycete sample bacillus and its application in mud decrement - Google Patents

One plant of pseudomycete sample bacillus and its application in mud decrement Download PDF

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Publication number
CN110218682A
CN110218682A CN201910559162.1A CN201910559162A CN110218682A CN 110218682 A CN110218682 A CN 110218682A CN 201910559162 A CN201910559162 A CN 201910559162A CN 110218682 A CN110218682 A CN 110218682A
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bacillus
sludge
pseudomycete
pseudomycete sample
bacterial strain
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CN110218682B (en
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蔡天明
陈立伟
于洪霞
张琦
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Nanjing Agricultural University
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    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F11/00—Treatment of sludge; Devices therefor
    • C02F11/02—Biological treatment
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F11/00—Treatment of sludge; Devices therefor
    • C02F11/12—Treatment of sludge; Devices therefor by de-watering, drying or thickening
    • 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
    • C12N1/20—Bacteria; Culture media therefor
    • 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
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2303/00—Specific treatment goals
    • C02F2303/06—Sludge reduction, e.g. by lysis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Hydrology & Water Resources (AREA)
  • Environmental & Geological Engineering (AREA)
  • Water Supply & Treatment (AREA)
  • Molecular Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Treatment Of Sludge (AREA)

Abstract

The invention discloses one plant of pseudomycete sample bacillus, its classification naming is pseudomycete sample bacillus (Bacillus pseudomycoides), bacterial strain WNJL-1, China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on October 22nd, 2018, deposit number is CGMCC No.16602.The invention also discloses application of the above-mentioned pseudomycete sample bacillus in mud decrement.Bacterial strain of the invention is screened from chemical engineering sewage treatment plant secondary settling tank sludge and is obtained, and meets bio-safety regulation；The bacterial strain can crack sludge cell thallus, and the organic matters such as the polysaccharide discharged after the cell cracking that can degrade and protein, to achieve the purpose that realize sludge over dry substance decrement.

Description

One plant of pseudomycete sample bacillus and its application in mud decrement
Technical field
The invention belongs to microorganisms technical fields, and in particular to a kind of pseudomycete sample bacillus and its in mud decrement Application.
Background technique
Excess sludge is the Main By product of Sewage Biological Treatment.Under normal circumstances, the yield of excess sludge is with volume For about sewage volume handled by municipal sewage plant 0.3%-0.5%, on the other hand with ten thousand tons of waste water for quality The dry weight for generating excess sludge is about 2.7 tons, it can be seen that the yield of excess sludge is huge.Waste water advanced place is carried out some The sewage treatment plants of reason, the yield of sludge can also further increase 0.5-1 times.According to live build portion's data, the end of the year 2010 National cities and towns dewatered sludge yield is close to 22,000,000 tons, this numerical value increases 35,600,000 tons within 2015.If pressing above-mentioned residue If sludge yield is the 0.3%-0.5% meter of sewage quantity, the surplus sludge volume in the year two thousand twenty China will reach 5.21-8.68 × 107 Ton or so.
What Sludge Reduction Technologies exactly came into being in this context.Sludge Reduction Technologies refer to and are guaranteeing at sewage Under the premise of managing effect, the various technologies for handling the reduction of sludge quantity caused by same amount of sewage are made using adequate measures. Existing method includes physics decrement, chemistry decrement, biology decrement.Compared with physics, chemical Subtraction method, biological Decrement Technique exists Economical rationality, easy to operate, environmental protection and energy saving corrosion-free to equipment etc. show biggish advantage, therefore sludge organism is reduced Technology is current research hotspot and development trend.
Summary of the invention
Technical problem to be solved by the invention is to provide the biologies that a kind of pseudomycete sample bacillus can be applied to sludge Decrement.
The present invention also technical problems to be solved are to provide above-mentioned strain in excess sludge reduction and improve dehydration property The application of energy aspect.
In order to solve the above technical problems, The technical solution adopted by the invention is as follows:
One plant of pseudomycete sample bacillus, classification naming are pseudomycete sample bacillus (Bacillus Pseudomycoides), bacterial strain WNJL-1 has been preserved in Chinese microorganism strain preservation management committee on October 22nd, 2018 Member's meeting common micro-organisms center, deposit number are CGMCC No.16602, depositary institution address are as follows: Chaoyang District, Beijing City North Star west The institute 3 of road 1.
Above-mentioned bacterial strains WNJL-1 is that in August, 2017 is screened from Nanjing Sewage Disposal sludge concentration tank One plant of obtained pseudomycete sample bacillus.The biological property of above-mentioned bacterial strains WNJL-1 is that bacterium colony is long filament dress, thallus stock Shape has gemma, sees Fig. 1.Gram-positive (Fig. 2), oxidase positive contact enzyme positive, and indoles is negative, can hydrolyze starch.It surveys 16S rDNA major part sequence is obtained, as shown in SEQ ID No:1.Column will be sequenced and carry out base sequence with the BLAST of the website NCBI The comparison of column, the results showed that bacterial strain and pseudomycete sample bacillus (Bacillus pseudomycoides) homology highest, Reach 99.79%.
A kind of pseudomycete sample gemma bacillus agent contains above-mentioned pseudomycete sample bacillus.
The LB Liquid Culture after improvement is added in bacterial strain WNJL-1 by the preparation method of above-mentioned pseudomycete sample gemma bacillus agent The bacterial suspension that base is formed.
Wherein, the LB liquid medium formula after the improvement be peptone 8g, NaCl 8g, yeast powder 4.0g, go from Sub- water 1000mL, pH 6.0.
Application of the above-mentioned pseudomycete sample bacillus in excess sludge reduction and/or dehydration is also in protection model of the invention Within enclosing.
Wherein, the excess sludge initial concentration be 5000mg/L~15000mg/L, the excess sludge it is initial PH is 6~8.
Specific application method is to mix pseudomycete sample gemma bacillus agent with 1: 50 volume ratio with excess sludge, bacterium Strain WNJL-1 dosing mass ratio is 0.01wt%~0.03wt%, 20 DEG C~40 DEG C conditions according to dry bacterium weight/wet mud re-computation Lower aeration 72-96h.
The invention has the advantages that:
(1) bacterial strain is suitable for growing in activated Sludge System, can be applied directly in biochemical wastewater treatment system to improve system The ability of system processing sewage, and the generation of sludge quantity in system is reduced simultaneously, it can also be added directly and be carried out into mudpan Aerobic biodegradation realizes mud decrement.
(2) its principle: 1) by biological lysis, constantly decomposing the microbial cells progress of excess sludge again, from And the yield of excess sludge is reduced, realize sludge in-situ decrement.2) by having to the polysaccharide and protein etc. that are discharged after cracking Machine object is degraded, and realizes that sludge over dry substance decrement and dewatering improve.
Detailed description of the invention
Growthform of Fig. 1 bacterial strain WNJL-1 on LB solid medium.
Fig. 2 bacterial strain WNJL-1 violet staining photo.
Influence of Fig. 3 microbial inoculum dosage to mud decrement treatment effect.
Fig. 4 microbial inoculum adds the sludge specific resistance variation diagram after processing 72h.
Specific embodiment
According to following embodiments, the present invention may be better understood.However, as it will be easily appreciated by one skilled in the art that real It applies content described in example and is merely to illustrate the present invention, without sheet described in detail in claims should will not be limited Invention.
In following embodiment, detection method is as follows:
(1) bacterial concentration measures: taking culture bacterium solution to measure its OD600 value, while using gravimetric detemination bacterium dry weight, and roll over Calculating is mycoplasma amount (mg/L) in every liter；
(2) handle the measurement of Sludge Property effect: during Treatment of Sludge every timing sampling for 24 hours to the MLSS of sludge, MLVSS, SV30 and sludge specific resistance.
Sludge MLSS is defined as sludge suspension solid concentration, unit mg/L.
Sludge MLVSS is defined as sludge volatile suspended solids concentration, unit mg/L.
Sludge SV30 is defined as: mud mixed liquid is stirred evenly, 100mL mixed liquor is taken to be placed in 100mL cleaning graduated cylinder, The volume that precipitating sludge formed in free settling process is recorded after standing 30min accounts for the percentage of former mixed liquor volume.
Sludge specific resistance is defined as: the resistance when sludge of position quality filters under a certain pressure in unit filter area Power.
Embodiment 1: bacteria fermentation production
The original seed of pseudomycete sample bacillus WNJL-1 of the invention is activated on LB solid medium, is inoculated in test tube It is spare on inclined-plane.Test tube species are inoculated in the culture medium of LB containing 200ml (LB culture medium prescription g/L: yeast extract 4.00, peptone 8.00, NaCl 8.00, pH6.0,1000ml shaking flask in, constant-temperature shaking culture to logarithmic phase, prepare inoculation seeding tank.Seed 500 liters of tank, 400 liters of inventory, culture medium prescription are as follows: glucose 1%, (NH4)2SO40.05%, K2HPO40.15%, MgSO40.01%, peptone 0.05%, pH 7.2-7.5.115 DEG C of high pressure moist heat sterilizations after feeding intake, after being cooled to 30 DEG C, Above-mentioned cultured shaking flask strain is accessed into 500 liters of seeding tanks, culture to logarithmic growth phase, mixing speed by 10% inoculum concentration It is 180 revs/min, filtrated air intake is 1: 0.8.The seed liquor for being up to logarithmic growth phase is given birth to by 10% inoculum concentration access Tank culture is produced, tank used medium ingredient is produced and seed tank culture base phase is same.5 tons of tankage size, 4.5 tons of inventory of production.It throws Tank 1.1kg/cm is produced after material2Pressure under, 115 DEG C of high pressure moist heat sterilizations are cooled to 30 DEG C hereinafter, logical filtrated air after sterilizing Keep germ-free condition spare.Production tank temperature degree after inoculation is controlled at 30 DEG C, produces the logical of filtrated air in the incubation of tank Tolerance is 1: 0.8-1.0, and mixing speed is 180 revs/min, and entire process flow incubation time is 48-60 hours.After fermentation Thalline quantity reaches 1,000,000,000/ml or more.
Embodiment 2: pseudomycete sample bacillus WNJL-1 handles Sludge Property measurement
Take uniformly mixed secondary settling tank excess sludge (saving at 11215mg/L, pH=7.46,4 DEG C of sludge concentration) 200mL is added in 250mL conical flask, and merging constant-temperature table activates for 24 hours in 28 DEG C, under the conditions of 150r/min, throws by 0.02wt% After pseudomycete sample bacillus WNJL-1 bacteria suspension is added in dosage, with breathing freely, sterile sealed membrane is sealed, and makes it at 28 DEG C, 150r/ Aerobic biodegradation is carried out in the constant-temperature table of min, during which no longer adds any nutrients, using sludge as sole carbon source, and Same period blank control is set.Every timing sampling for 24 hours to MLSS, SV of sludge during culture30, centrifugal dehydration rate is measured. The performance test results are that sludge MLSS removal rate reaches peak after pseudomycete sample bacillus WNJL-1 handles 72h 20.18%, control group sludge MLSS removal rate is 2.6% at this time；Sludge MLVSS removal rate reaches 28.35%；Sludge SV30By Initial value 93.5% drops to 60.8%, and control group sludge (sludge that control group is not inoculating strain WNJL-1) SV30 is at this time 78.5%.
Influence of the 3 microbial inoculum dosage of embodiment to mud decrement treatment effect
Take uniformly mixed secondary settling tank excess sludge (saving at 11215mg/L, pH=7.46,4 DEG C of sludge concentration) 200mL is added in 250mL conical flask, and merging constant-temperature table activates for 24 hours in 28 DEG C, under the conditions of 150r/min, presses respectively Pseudomycete sample bacillus is added in 0.01wt%, 0.02wt%, 0.03wt%, 0.04wt% and 0.05wt% (v/v) dosage After WNJL-1 bacteria suspension, with breathing freely, sterile sealed membrane is sealed, it is made to carry out aerobic life in 28 DEG C, the constant-temperature table of 150r/min Object degradation, during which no longer adds any nutrients, using sludge as sole carbon source, and same period blank control is arranged.MLSS after 72h Fig. 3 is seen with MLVSS removal rate.As seen from Figure 3, when microbial inoculum dosage range is in the range of 0.01wt%-0.05wt% Interior, raising microbial inoculum adds ratio and can effectively improve the removal rate of MLSS and MLVSS in sludge in 72h, i.e. raising mud decrement Rate；Continue to improve and adds ratio to 8%.Removal rate does not obviously increase, and considers from operating cost angle, and 0.02wt% is Best microbial inoculum adds ratio.
Embodiment 4
Take uniformly mixed secondary settling tank excess sludge (saving at 11215mg/L, pH=7.46,4 DEG C of sludge concentration) 200mL is added in 250mL conical flask, and merging constant-temperature table activates for 24 hours in 28 DEG C, under the conditions of 150r/min, presses respectively After pseudomycete sample bacillus WNJL-1 bacteria suspension is added in 0.02wt% dosage, with breathing freely, sterile sealed membrane is sealed, make its 28 DEG C, aerobic biodegradation is carried out in the constant-temperature table of 150r/min, any nutrients is during which no longer added, using sludge as only One carbon source, and same period blank control is set.Fig. 4 is shown in sludge specific resistance variation after 72h post-processing.After handling as seen from Figure 4 Sludge specific resistance be decreased obviously, plate and frame filter press can be directly entered and be carried out dehydrating.
Sequence table
<110>Agricultural University Of Nanjing
<120>one plants of pseudomycete sample bacillus and its application in mud decrement
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1451
<212> DNA
<213>pseudomycete sample bacillus (Bacillus pseudomycoides)
<400> 1
tggattgcgg tgctatacat gcagtcgagc gaatggatta agagcttgct cttatgaagt 60
tagcggcgga cgggtgagta acacgtgggt aacctgccca taagactggg ataactccgg 120
gaaaccgggg ctaataccgg ataacatttt gcaccgcatg gtgcgaaatt caaaggcggc 180
ttcggctgtc acttatggat ggacccgcgt cgcattagct agttggtgag gtaacggctc 240
accaaggcaa cgatgcgtag ccgacctgag agggtgatcg gccacactgg gactgagaca 300
cggcccagac tcctacggga ggcagcagta gggaatcttc cgcaatggac gaaagtctga 360
cggagcaacg ccgcgtgagt gatgaaggct ttcgggtcgt aaaactctgt tgttagggaa 420
gaacaagtgc tagttgaata agctggcacc ttgacggtac ctaaccagaa agccacggct 480
aactacgtgc cagcagccgc ggtaatacgt aggtggcaag cgttatccgg aattattggg 540
cgtaaagcgc gcgcaggtgg tttcttaagt ctgatgtgaa agcccacggc tcaaccgtgg 600
agggtcattg gaaactggga gacttgagtg cagaagagga aagtggaatt ccatgtgtag 660
cggtgaaatg cgtagagata tggaggaaca ccagtggcga aggcgacttt ctggtctgta 720
actgacactg aggcgcgaaa gcgtggggag caaacaggat tagataccct ggtagtccac 780
gccgtaaacg atgagtgcta agtgttagag ggtttccgcc ctttagtgct gaagttaacg 840
cattaagcac tccgcctggg gagtacggcc gcaaggctga aactcaaagg aattgacggg 900
ggcccgcaca agcggtggag catgtggttt aattcgaagc aacgcgaaga accttaccag 960
gtcttgacat cctctgacaa ccctagagat agggcttccc cttcgggggc agagtgacag 1020
gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc 1080
gcaacccttg atcttagttg ccatcattaa gttgggcact ctaaggtgac tgccggtgac 1140
aaaccggagg aaggtgggga tgacgtcaaa tcatcatgcc ccttatgacc tgggctacac 1200
acgtgctaca atggacggta caaagagctg caagaccgcg aggtggagct aatctcataa 1260
aaccgttctc agttcggatt gtaggctgca actcgcctac atgaagctgg aatcgctagt 1320
aatcgcggat cagcatgccg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca 1380
caccacgaga gtttgtaaca cccgaagtcg gtggggtaac cttttggagc cagccgctaa 1440
gtgacagaag g 1451

Claims (7)

1. one plant of pseudomycete sample bacillus, classification naming is pseudomycete sample bacillus (Bacillus Pseudomycoides), bacterial strain WNJL-1 has been preserved in Chinese microorganism strain preservation management committee on October 22nd, 2018 Member's meeting common micro-organisms center, deposit number are CGMCC No.16602.
2. a kind of pseudomycete sample gemma bacillus agent, which is characterized in that contain pseudomycete sample gemma bar described in claim 1 Bacterium.
3. the preparation method of pseudomycete sample gemma bacillus agent as claimed in claim 2, which is characterized in that add bacterial strain WNJL-1 The bacterial suspension formed after LB liquid medium fermentation after entering improvement.
4. preparation method according to claim 3, which is characterized in that the LB liquid medium formula after the improvement is Peptone 8g, NaCl 8g, yeast powder 4.0g, deionized water 1000mL, pH 6.0.
5. application of the pseudomycete sample bacillus described in claim 1 in excess sludge reduction and/or dehydration.
6. application according to claim 5, which is characterized in that the excess sludge initial concentration be 5000mg/L~ 15000mg/L, the initial pH of the excess sludge are 6~8.
7. application according to claim 5, which is characterized in that by pseudomycete sample gemma bacillus agent and excess sludge with 1: The mixing of 50 volume ratios, bacterial strain WNJL-1 dosing mass ratio are 0.01wt%~0.03wt% according to dry bacterium weight/wet mud re-computation, 72-96h is aerated under the conditions of 20 DEG C~40 DEG C.
CN201910559162.1A 2019-06-26 2019-06-26 Pseudomycosis bacillus and application thereof in sludge reduction Active CN110218682B (en)

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Cited By (3)

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CN114456982A (en) * 2022-03-03 2022-05-10 青岛蔚蓝赛德生物科技有限公司 Bacillus brevis and application thereof in degradation or digestion of sludge
CN114703094A (en) * 2022-03-18 2022-07-05 山东野田生物科技有限公司 Bacillus pseudomycoides and application thereof in preparation of plant pathogenic bacteria bacteriostat
CN115960777A (en) * 2022-12-06 2023-04-14 江苏省中国科学院植物研究所 Pseudomycobacillus sp CNBG-PGPR-20 and application thereof in prevention and treatment of vegetable epidemic diseases

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HASSIMI ABU HASAN 等: "Removal of ibuprofen, ketoprofen, COD and nitrogen compounds from pharmaceutical wastewater using aerobic suspension-sequencing batch reactor (ASSBR)", 《SEPARATION AND PURIFICATION TECHNOLOGY》 *
胡怡杉 等: "MBR和CAS工艺污泥在贫营养培养条件下的微生物群落结构研究", 《环境科学学报》 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114456982A (en) * 2022-03-03 2022-05-10 青岛蔚蓝赛德生物科技有限公司 Bacillus brevis and application thereof in degradation or digestion of sludge
CN114456982B (en) * 2022-03-03 2023-09-26 青岛蔚蓝赛德生物科技有限公司 Brevibacillus brevis and application thereof in degrading or digesting sludge
CN114703094A (en) * 2022-03-18 2022-07-05 山东野田生物科技有限公司 Bacillus pseudomycoides and application thereof in preparation of plant pathogenic bacteria bacteriostat
CN114703094B (en) * 2022-03-18 2023-09-05 山东野田生物科技有限公司 Bacillus pseudomycosis and application thereof in preparation of plant pathogenic bacteria bacteriostat
CN115960777A (en) * 2022-12-06 2023-04-14 江苏省中国科学院植物研究所 Pseudomycobacillus sp CNBG-PGPR-20 and application thereof in prevention and treatment of vegetable epidemic diseases
CN115960777B (en) * 2022-12-06 2023-09-12 江苏省中国科学院植物研究所 Bacillus pseudomycoides and application thereof in prevention and treatment of vegetable epidemic disease

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