CN114874938B - Bacillus cereus S5 for degrading hydrogen sulfide gas and application thereof - Google Patents
Bacillus cereus S5 for degrading hydrogen sulfide gas and application thereof Download PDFInfo
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- CN114874938B CN114874938B CN202210498523.8A CN202210498523A CN114874938B CN 114874938 B CN114874938 B CN 114874938B CN 202210498523 A CN202210498523 A CN 202210498523A CN 114874938 B CN114874938 B CN 114874938B
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- bacillus cereus
- hydrogen sulfide
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- 241000193755 Bacillus cereus Species 0.000 title claims abstract description 33
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 title claims abstract description 20
- 229910000037 hydrogen sulfide Inorganic materials 0.000 title claims abstract description 19
- 230000000593 degrading effect Effects 0.000 title claims abstract description 16
- 239000007789 gas Substances 0.000 title claims abstract description 14
- 238000004321 preservation Methods 0.000 claims abstract description 3
- 241001660259 Cereus <cactus> Species 0.000 claims abstract 2
- -1 sulfide ions Chemical class 0.000 claims description 9
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 abstract description 28
- 230000015556 catabolic process Effects 0.000 abstract description 14
- 238000006731 degradation reaction Methods 0.000 abstract description 14
- 230000000694 effects Effects 0.000 abstract description 9
- 238000000034 method Methods 0.000 abstract description 9
- 210000003608 fece Anatomy 0.000 abstract description 8
- 239000010871 livestock manure Substances 0.000 abstract description 8
- 239000002361 compost Substances 0.000 abstract description 7
- 238000009264 composting Methods 0.000 abstract description 7
- 244000144972 livestock Species 0.000 abstract description 6
- 244000144977 poultry Species 0.000 abstract description 6
- 239000001963 growth medium Substances 0.000 description 12
- 238000012216 screening Methods 0.000 description 11
- 239000011593 sulfur Substances 0.000 description 8
- 229910052717 sulfur Inorganic materials 0.000 description 8
- 239000007788 liquid Substances 0.000 description 7
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 6
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 5
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 230000001651 autotrophic effect Effects 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 238000006477 desulfuration reaction Methods 0.000 description 4
- 230000023556 desulfurization Effects 0.000 description 4
- 230000003009 desulfurizing effect Effects 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 150000004763 sulfides Chemical class 0.000 description 3
- 239000012855 volatile organic compound Substances 0.000 description 3
- 241000605118 Thiobacillus Species 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 235000019270 ammonium chloride Nutrition 0.000 description 2
- 238000010170 biological method Methods 0.000 description 2
- 150000004770 chalcogenides Chemical class 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 229940050906 magnesium chloride hexahydrate Drugs 0.000 description 2
- DHRRIBDTHFBPNG-UHFFFAOYSA-L magnesium dichloride hexahydrate Chemical compound O.O.O.O.O.O.[Mg+2].[Cl-].[Cl-] DHRRIBDTHFBPNG-UHFFFAOYSA-L 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 229920001021 polysulfide Polymers 0.000 description 2
- 239000005077 polysulfide Substances 0.000 description 2
- 150000008117 polysulfides Polymers 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- 229910052979 sodium sulfide Inorganic materials 0.000 description 2
- GRVFOGOEDUUMBP-UHFFFAOYSA-N sodium sulfide (anhydrous) Chemical compound [Na+].[Na+].[S-2] GRVFOGOEDUUMBP-UHFFFAOYSA-N 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 description 1
- 108020000946 Bacterial DNA Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical compound OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 1
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 description 1
- 241001509286 Thiobacillus denitrificans Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006065 biodegradation reaction Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000005059 dormancy Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000004630 mental health Effects 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- XTQHKBHJIVJGKJ-UHFFFAOYSA-N sulfur monoxide Chemical class S=O XTQHKBHJIVJGKJ-UHFFFAOYSA-N 0.000 description 1
- 229910052815 sulfur oxide Inorganic materials 0.000 description 1
- DHCDFWKWKRSZHF-UHFFFAOYSA-N sulfurothioic S-acid Chemical compound OS(O)(=O)=S DHCDFWKWKRSZHF-UHFFFAOYSA-N 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 210000002268 wool Anatomy 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
- B01D53/46—Removing components of defined structure
- B01D53/48—Sulfur compounds
- B01D53/52—Hydrogen sulfide
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
- B01D53/74—General processes for purification of waste gases; Apparatus or devices specially adapted therefor
- B01D53/84—Biological processes
-
- 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
Landscapes
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Health & Medical Sciences (AREA)
- Environmental & Geological Engineering (AREA)
- Analytical Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
The invention belongs to the technical field of livestock and poultry manure harmless treatment, and particularly discloses bacillus cereus S5 for degrading hydrogen sulfide gas and application thereof. The invention screens out a bacillus cereus strainBacillus cereus) S5, the preservation number is: the invention screens out odor degrading bacteria, which can effectively degrade hydrogen sulfide gas generated in the composting process and achieve the effect of purifying compost odor. Good degradation effect and simple operation, and provides an effective method for treating compost odor.
Description
Technical Field
The invention belongs to the technical field of livestock and poultry manure harmless treatment, and particularly relates to bacillus cereus S5 for degrading hydrogen sulfide gas and application thereof.
Background
With the expansion of the livestock and poultry cultivation scale and the enhancement of environmental awareness, the odor problem in the cultivation process is more and more emphasized. The livestock and poultry raising scale is continuously increased, and a large amount of livestock and poultry manure is produced. The composting technology is one of important technologies for livestock and poultry manure treatment, and has the characteristic of environmental friendliness. However, a large amount of malodorous substances are often generated during composting. Ammonia, hydrogen sulfide and most Volatile Organic Compounds (VOCs) generated during composting cause pollution of the air environment and affect physical and mental health of surrounding residents.
The primary means for treating odor problems in composting plants is by biological methods. The biological method has good removal effect on ammonia, hydrogen sulfide and water-soluble malodorous substances in the composting process. However, the handling performance of bacteria-based bioreactors is generally poor for highly hydrophobic VOCs produced in compost. This is because such contaminants have low mass transfer efficiency between the gas and liquid phases, severely affecting the subsequent utilization of them by the bacteria.
In the microbial oxidation of H 2 In the S process, microorganisms for biodegradation are called as sulfur-removing bacteria, namely sulfur bacteria, and two major types of autotrophic bacteria and heterotrophic bacteria exist, wherein the autotrophic bacteria have strong capability of synthesizing organic matters and are beneficial to the conversion of inorganic matters. There are many bacteria reported in the literature to decompose sulfur and sulfides, and the bacteria that obtain energy from sulfur oxides and sulfides are mainly chalcogenides and chalcogenides.
The genus Thiobacillus (Thiobacillus) is a small bacillus, monopolar wool exercise, no dormancy stage, gram positive. The catalyst is mainly used for oxidizing sulfides, including sulfite, elemental sulfur, thiosulfate, polysulfide and the like. The oxidation results in sulfate production and extracellular accumulation of sulfur particles and polysulfide compounds. The pH range of the seed is wide, the optimum temperature is 28-30 ℃, and the seed is aerobic except for thiobacillus denitrificans. The bacteria belong to three types of autotrophic bacteria, autotrophic bacteria and heterotrophic bacteria.
The bacillus cereus can produce antibacterial substances, inhibit the reproduction of harmful microorganisms, degrade nutrient components in soil and improve ecological environment. At present, there is no report on degradation of hydrogen sulfide gas by bacillus cereus.
Disclosure of Invention
The invention aims to provide Bacillus cereus S5 for degrading hydrogen sulfide gas in compost, and the preservation number of the strain is as follows: cctccc No. M2022347.
Another object of the invention is to provide the use of bacillus cereus S5 for degrading hydrogen sulphide gas.
In order to achieve the above object, the present invention adopts the following technical measures:
acquisition of Bacillus cereus S5:
the Bacillus cereus S5 is obtained by separating, screening and identifying fillers of a biotrickling filter which is used for degrading ammonia gas and hydrogen sulfide gas and runs stably for a long time, extracting bacterial DNA, amplifying a 16SrDNA sequence by PCR, submitting an NCBI database for comparison, and determining the Bacillus cereus by referring to a Bojie system bacteriology handbook and a common bacterial system identification handbook, wherein the Bacillus cereus accords with physiological and biochemical characteristics of the Bacillus cereus. Bacillus cereus is a gram positive and milky white colony, the colony size is 25-108 mm, the colony is in the shape of a moniliform, the edge is neat, and the optimal growth temperature is 37 ℃.
The strain was sent to China center for type culture Collection, and was named after classification, at 3.30 of 2022: bacillus cereus S5, accession number: CCTCC NO: M2022347, address: university of martial arts in chinese.
The application of bacillus cereus S5 in degrading sulfide ions, in particular to degradation of hydrogen sulfide gas.
Compared with the prior art, the invention has the following advantages:
the invention screens out odor degrading bacteria, can effectively degrade hydrogen sulfide gas generated in the composting process, and achieves the effect of purifying compost odor. The method has good degradation effect and simple operation, and provides an effective method for degrading compost odor.
Drawings
FIG. 1 is a schematic diagram showing the screening results of the sulfur-removing bacteria;
wherein: a) Measuring the degradation capacity of the sulfur removal bacteria; b) Degradation rate of sulfide ion.
FIG. 2 is S 2- Content variation curve and S 2- Degradation rate change curve.
Fig. 3 is a schematic diagram of an effect test of bacillus cereus S5 on degrading hydrogen sulfide in pig manure.
Detailed description of the preferred embodiments
In the technical scheme of the invention, the reagents are purchased from a biochemical store unless specifically indicated, and the technical scheme is a conventional technology in the field unless specifically indicated.
Example 1:
separation, purification and identification of bacillus cereus S5:
the applicant separates, screens and identifies the filler in the biological trickling filter which is used for degrading ammonia gas and hydrogen sulfide gas and stably operates for a long time. The specific separation and purification method comprises the following steps: weighing compost sample 5g in 150mL conical flask, adding water to 100mL shaking table 180r/min and shaking at 37deg.C for 1 hr, collecting supernatant 2mL, inoculating into desulfurizing bacteria screening culture medium, standing at 30deg.C for culturing in 100mL/250mL system, and detecting S 2- After the ion consumption is finished, 5mL of enrichment culture solution is transferred to fresh enrichment culture solution, and continuous enrichment transfer is carried out for 4 times. Culturing the culture solution in the final screening culture medium on a solid plate with corresponding components in a constant temperature incubator at 30 ℃ for 3-5 d, picking out bacterial colonies with different characteristics, separating by streaking, and storing in a refrigerator at 4 ℃ for later use.
Re-inoculating the strain with different characteristics which can grow in the desulfurizing bacteria screening culture medium, standing at 30deg.C in 100mL/250mL system, culturing for 48 hr, repeating for 3 times, and selecting S 2- The strain with the highest ion degradation rate.
Desulfurizing bacteria screening culture medium: 2.0g of monopotassium phosphate, 0.4g of ammonium chloride, 0.4g of sodium carbonate, 0.2g of magnesium chloride hexahydrate, 1000mL of distilled water and pH 7.0. Substrate sodium sulfide is added into the culture medium, S 2- The initial concentration was set at 500mg/L.
After enrichment culture, 8 strains which have different characteristics and can grow on a desulfurization bacteria screening culture medium are obtained through separation, and the strains are named S1-S8 in sequence. Inoculating 8 strains of bacteria to a desulfurization bacteria screening culture medium, and measuring the degradation effect of 8 strains of bacteria on sulfur ions after shake culture for 48 hours in a shaking table. The effect of 8 strains on degrading sulfide ions is shown in figure 1. After 48h of culture, the average content of sulfide ions in the strain S5 group is 71.33mg/L, and the degradation rate of the strain S5 group can reach 74.6% compared with the degradation rate of the strain CK group; the sulfur ion degradation rates of the other test groups S3 and S7 were 10.69% and 10.75%, respectively. The strain S5 has significantly stronger degradability to sulfide ions than other test groups.
Extracting DNA of bacteria S5, amplifying a 16SrDNA sequence by PCR, submitting the sequence to NCBI database for comparison, and determining the bacillus cereus by referring to the physiological and biochemical characteristics of bacillus cereus in Bojie' S system bacteriology handbook and the common bacterial system identification handbook.
Bacillus cereus is a gram positive white colony, a milky white colony, the colony size is 25-108 mm, the bacillus cereus is in the shape of a moniliform, the edge is neat, and the optimal growth temperature is 37 ℃.
The strains are sent to China center for type culture Collection (China) for 3 months and 30 days in 2022, and are named after classification: bacillus cereus S5, accession number: CCTCC NO: M2022347, address: university of martial arts in chinese.
Example 2:
bacillus cereus S5 fermentation culture:
resuscitating Bacillus cereus, inoculating 100 μ L S5 bacteria liquid into 5mL LB liquid medium, culturing at 37deg.C with shaking table 180r/min for 12 hr to stationary phase, and counting to obtain effective bacteria concentration of 2.5X10 8 cfu/ml。
Example 3:
bacillus cereus S5 pair S 2- Degradation of ions:
desulfurizing bacteria screening culture medium
2.0g of monopotassium phosphate, 0.4g of ammonium chloride, 0.4g of sodium carbonate, 0.2g of magnesium chloride hexahydrate and 1000mL of distilled water are weighed, the pH is regulated to 7+/-0.05, and the mixture is autoclaved for 20min at 121 ℃. Adding substrate sodium sulfide into the culture medium after cooling the temperature, S 2- The initial concentration was set at 500mg/L.
Inoculation of bacterial liquid
Inoculating bacillus cereus into the desulfurization screening culture medium, and setting a blank control, wherein the blank control is the desulfurization screening culture medium without inoculating microorganisms. And respectively measuring the sulfur ion content in the culture medium at 2h, 4h, 6h, 12h, 24h and 48 h.
Calculation of the Sulfur ion degradation Rate
RE％＝(C CK -C S5 )/C CK ×100％
C CK -blank groupS 2- Content of
C S5 S of group S5 of microbial agents 2- Content of
The results are shown in FIG. 2, and the specific values are as follows:
| 0h | 2h | 4h | 6h | 12h | 24h | 48h | |
| S5(mg/L) | 500.00 | 280.33 | 252.50 | 245.50 | 228.50 | 201.33 | 71.33 |
| CK(mg/L) | 500.00 | 289.50 | 285.00 | 283.67 | 283.50 | 283.67 | 269.83 |
| RE(％) | - | 3.17 | 11.4 | 13.46 | 19.4 | 29.03 | 72.56 |
example 4:
effect test of bacillus cereus S5 for removing hydrogen sulfide in pig manure
200g of fresh pig manure was placed in a 1000mL beaker, and Bacillus cereus S5 was inoculated in the beaker in an inoculum size of 5%. Uniformly mixing bacterial liquid and pig manure by using a glass rod, filling a small built-in beaker with 20mL of hydrogen sulfide absorption liquid, sealing by using a preservative film, supplementing an equal amount of sterile water to a control group, repeating each group for 3 times, taking the small beaker every 2 days to measure hydrogen sulfide in the absorption liquid, replacing the absorption liquid, measuring the release amount of the hydrogen sulfide by using a methylene blue spectrophotometry, and performing difference significance analysis with a blank control group.
The test results are shown in the following table:
Claims (3)
1. isolated bacillus cereusBacillus cereus) The preservation number of the bacillus cereus is as follows: cctccc No. M2022347.
2. The use of bacillus cereus of claim 1 for degrading negative divalent sulfide ions.
3. The use of bacillus cereus of claim 1 for degrading hydrogen sulfide gas.
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| CN106676046A (en) * | 2017-02-21 | 2017-05-17 | 宁波大学 | Bacillus cereus and application thereof in oxidizing inorganic sulfide |
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