CN113736686B - Microbial deodorant for desulfurizing and deaminizing and preparation method thereof - Google Patents
Microbial deodorant for desulfurizing and deaminizing and preparation method thereof Download PDFInfo
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Classifications
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- 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
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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
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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
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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
- C05F3/00—Fertilisers from human or animal excrements, e.g. manure
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/101—Sulfur compounds
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
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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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
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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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Abstract
The present invention relates to a microbial composition, and more particularly, to a microbial composition having deodorizing effect. Comprises Bacillus altitudinis (GAU-00532) and bacillus subtilis Bacillus subtilis subsp.Stercoris (GAU-00533) and Bacillus zhangzhouensis (GAU-00534), wherein the three bacillus are preserved in China center for type culture Collection, and the preservation addresses are university of Wuhan in Wuhan, and the preservation numbers are CCTCC NO: M2021352, CCTCC NO: M2021350 and CCTCC NO: M2021351 respectively. The desulfurizing and deodorizing microbial deodorant disclosed by the invention has the advantages of low cost, less consumption, high deodorizing efficiency, no chemical additives and complex control systems, and the deodorizing products can be directly utilized without being treated again.
Description
Technical Field
The present invention relates to a microbial composition, and more particularly, to a microbial composition having deodorizing effect.
Background
With the increasing promotion of the living standard of people and the continuous expansion of the demand of meat food, the rapid development of livestock and poultry farming industry is promoted, but most farms do not have perfect manure treatment facilities, so that not only are the bad smell of surrounding environment, sewage flowing and breeding of mosquitoes and flies caused, but also the pollution of organic matters and pathogenic bacteria of soil and water is more serious. The odor of pig farms mainly comes from the smell of pig bodies, the gas discharged from respiratory tracts and the excreted fecaluria of bodies, which are the main sources of malodorous gases, contains undigested organic substances such as starch, fat, protein, non-starch carbohydrate and the like, and can generate malodorous compounds such as ammonia, sulfides (hydrogen sulfide, methyl mercaptan) and the like under the anaerobic or facultative anaerobic condition through the decomposition of microorganisms. In most cases, malodorous and offensive gas content increases gradually as the storage time of bodily wastes increases. High concentration ammonia gas can cause keratitis, conjunctivitis, dermatitis, colibacillosis and the like of pigs and poultry, while low concentration ammonia gas can cause growth and development failure, sexual maturity delay and the like, and seriously influence the production performance of pigs and poultry; the low concentration of hydrogen sulfide stimulates the nasal cavity, causes rhinitis, pulmonary edema, weak constitution and reduced immunity, and the high concentration of hydrogen sulfide causes pigs to show photophobia, tear, lose appetite and perception, and even die due to central nerve paralysis. Therefore, the method reduces the content of malodorous compounds such as ammonia, hydrogen sulfide and the like in livestock and poultry cultivation, is beneficial to improving the production performance of livestock and poultry, is beneficial to reducing the atmospheric pollution, solves the problem that the malodorous compounds cause complaints and disputes of surrounding residents, and is an important measure for controlling the environmental sanitation of farms and guaranteeing the healthy development of livestock and poultry cultivation.
In recent years, research at home and abroad shows that compared with a chemical method, the microbial deodorization method has low cost, no chemical additive and complex control system, and the deodorization product can be directly utilized without retreatment; compared with a physical method, the microbial deodorization does not need dilution or odor masking, but utilizes the physiological metabolism of microorganisms to decompose or eliminate malodorous compounds, has the advantages of high deodorization rate, no secondary pollution, safe use, good continuous effect, high economic benefit and the like, and meanwhile, the deodorized livestock and poultry manure can be used as an organic fertilizer to improve the soil aggregate structure, the organic matter content of the soil and the saline-alkali soil, inhibit crop diseases and insect pests, and becomes an important way for treating malodorous gases such as ammonia, hydrogen sulfide and the like in the current livestock and poultry cultivation.
Biological deodorization has been widely used, and the main methods are as follows:
1. biological filtration method
The biological filtration method, which is a mature biochemical method for treating odor, is a biological filtration method, namely, waste gas is introduced into a reactor filled with filter material attached microorganisms, and the microorganisms attached on the filter material are utilized to degrade volatile odor such as hydrogen sulfide. The odor is firstly absorbed by the filter material, and then is oxidized and decomposed by microorganisms attached to the filter material to form micromolecular substances, so that the odor is removed.
It follows that the choice of filter material is critical, and that a good quality filter material must have a large surface area for the growth of microorganisms while meeting the growth of a variety of microorganisms; has better adsorption performance and fully absorbs odor substances. In addition to this, a large porosity and a better water absorption are required.
2. Biological washing method
The main principle of the biological washing method (also called as a biological absorption method) is that the main component of the odor is transferred into a liquid phase, and then the wastewater with the odor component absorbed is treated by an activated sludge method, and the biological washing method according to a gas-liquid contact mode currently has two methods, namely an aeration type and a washing type. The aeration biological washing method is similar to the activated sludge method, except that aeration air is replaced by malodorous gas in a sewage treatment station, and compared with the common equipment, the aeration tank is adopted, the malodorous gas is fully contacted with the activated sludge through the aeration tank, and the malodorous gas is removed through the metabolism of microorganisms in newcastle. The method is suitable for treating odor substances such as hydrogen sulfide, lower alcohol and the like.
The washing type deodorization method is generally divided into two parts, namely an absorption tower and an activated sludge tower, the main function of the absorption tower is to transfer the main components of odor into a liquid phase, thereby realizing a mass transfer process, and then the part of wastewater with the odor components absorbed is treated by the activated sludge method through metabolism of microorganisms. In contrast to the biofiltration process, the liquid phase of the bioscouring process is not stationary but is mobile. Thus, the reaction conditions can be well controlled, and the addition of buffering agents and the like is facilitated.
At present, most of the microbial deodorant developed at home and abroad is a mixed bacterial system formed by fungi and bacteria according to a certain proportion, the possibility of spore mutation of the fungi is quite high in the process of expanding the culture stage, and the growth of the bacteria can be inhibited due to the fact that the individual fungi are large, so that the properties of deodorizing products are changed, the quality is reduced, the deodorizing effect is affected, and even pathogenic bacteria are generated. The bacteria have the advantages of difficult mutation, easy culture, no pathogenicity of most saprophytic bacteria, strong adaptability to the bacteria and the like, and the high-efficiency deodorizing bacteria not only can improve the decomposition rate of malodorous substances, but also can effectively remove substances difficult to biochemically degrade.
In addition, the traditional screening method has poor directivity, and the deodorizing microorganisms are easy to generate variation, have poor environment adaptability and are easy to be interfered by other miscellaneous bacteria due to the limitations of the screening method, the specificity of the microorganisms and other reasons, and meanwhile, the deodorizing effect of the screened deodorizing microorganisms is not ideal. The invention adopts the directional enrichment domestication method, has the advantages of strong directivity during screening, difficult mutation of the screened deodorant microorganisms, strong anti-interference capability, obvious deodorant effect and the like, and effectively solves the problems faced by the traditional screening method.
Disclosure of Invention
In order to remove malodorous gases in small quantity and high efficiency, the invention provides various desulfurizing and ammonia-removing microbial deodorant and a preparation method thereof. The technical scheme is as follows:
a desulfurizing and ammonia-removing microbial deodorant is characterized by comprising Bacillus altitudinis (GAU-00532), bacillus subtilis Bacillus subtilis subsp. Stercoris (GAU-00533) and Bacillus zhangzhouensis (GAU-00534), wherein the three bacillus are preserved in China center for type culture Collection, and the preservation addresses are university of Wuhan in Wuhan, and the preservation numbers are CCTCC NO: M2021352, CCTCC NO: M2021350 and CCTCC NO: M2021351 respectively.
Wherein, in the desulfurization and deamination microbial deodorant, the ratio of Bacillus altitudinis (GAU-00532) to bacillus subtilis Bacillus subtilis subsp.
Or more preferably, the Bacillus altitudinis (GAU-00532), bacillus subtilis Bacillus subtilis subsp. Stercoris (GAU-00533) and Bacillus zhangzhouensis (GAU-00534) are cultured with LB medium at 30℃respectively, and then mixed.
The technical scheme provided by the invention has the beneficial effects that:
bacillus altitudinis (GAU-00532), bacillus subtilis (GAU-00533) and Bacillus zhangzhouensis (GAU-00534) are bacteria of the phylum Firmicutes, and have strong capability of resisting adverse environments such as high temperature, drought, barren nutrition and the like. Compared with chemical deodorization, the mixed desulfurizing and ammonia-removing microbial deodorant has the advantages of low cost, no chemical additives and complex control system, and the deodorizing products can be directly utilized without reprocessing.
Compared with a physical method, the compound microorganism deodorant obtained by the invention does not need dilution and odor masking, but utilizes the physiological metabolism of microorganisms to decompose or eliminate malodorous compounds, has the advantages of high deodorization rate, no secondary pollution, safe use, good continuous effect, high economic benefit and the like, and meanwhile, the deodorized livestock and poultry manure can be used as an organic fertilizer, thereby not only improving the soil aggregate structure, improving the soil organic matter content and improving the saline-alkali soil, but also inhibiting crop diseases and insect pests.
Compared with other similar microbial deodorants, the invention can remove hydrogen sulfide and ammonia. Has the advantages of low dosage and high deodorizing efficiency.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the present invention more apparent, the embodiments of the present invention will be described in further detail with reference to the accompanying drawings.
Embodiment one: bacterial screening
In order to obtain bacteria for high-efficiency desulfurization and ammonia removal and avoid the interference of other miscellaneous bacteria, pig manure compost and pig manure urine in different habitats are used as test materials, and oriented enrichment, domestication and desulfurization and ammonia removal microorganisms are carried out under the conditions of 28-35 ℃ and 160-220 r/min. Firstly, placing an acquired sample in an ammonia selective liquid culture medium with a certain concentration and a basic inorganic salt culture medium containing sodium sulfide, directionally enriching and domesticating the desulfurizing and ammonia-removing microorganisms, sequentially subculturing to 42 th generation, diluting and coating an enriched liquid on a Luria-Bertani culture medium (LB culture medium for short), picking single bacterial colonies, separating and purifying by a plate streaking method, and finally combining the single bacterial strains and screening the desulfurizing and ammonia-removing strains to obtain the high-efficiency desulfurizing and ammonia-removing bacterial strain.
Embodiment two: determination of deodorizing Performance of desulfurizing and deodorizing microbial deodorant
Bacillus altitudinis (GAU-00532) selected in example 1: bacillus subtilis Bacillus subtilis subsp. Stercoris (GAU-00533): bacillus zhangzhouensis (GAU-00534) and (0.8-1.2) and (1.6-2.5) are mixed according to the mass ratio of (1.0-1.8).
The test of the composite strain system shows that the 3 strains have no antagonistic reaction, the OD value is 0.764 after 12 hours of culture, and the increasing range is 38.61-51.96%; NH (NH) 4 + - (48 h) absorptivity is 79.8%, NH 4 + -an increase in the absorption rate of N (48 h) ranging from 16.79% to 20.18%; s is S 2- The yield of the adsorbent (48 h) is 60.5%, S 2- The yield of the adsorption (48 h) ranged from 29.09% -68.18% (Table 1).
Table 1: the main index content of the mixed strain system and the main index content of each strain before mixing are compared
Embodiment III: preparation and application of desulfurizing and deaminizing microbial deodorant
(1) Refrigerating strains through a freezing storage tube, activating respectively, performing primary expansion, performing secondary expansion, performing shaking table expansion, preparing, detecting, packaging liquid and obtaining the microbial deodorant; the specific preparation method comprises the following steps:
(1) microorganism selection, 3 microorganisms belonging to the genus Bacillus altitudinis (GAU-00532), bacillus subtilis Bacillus subtilis subsp. Stercoris (GAU-00533), bacillus zhangzhouensis (GAU-00534) were used as the pig farm microorganism deodorant.
(2) And (3) activating, namely taking the 3 bacterial frozen tube seeds out of a refrigerator at the temperature of minus 80 ℃, immediately placing the tube seeds in a water bath at the temperature of 38-40 ℃ for rapid recovery and proper shaking until the inner ice is completely dissolved, wherein 50-100 seconds are generally required. Under the aseptic condition, wiping the frozen tube by absorbent cotton soaked with 75% alcohol, sucking 50 mu L of proper strain by a liquid transfer device, dripping the strain near one edge of a solid LB culture medium, inoculating the strain to the culture medium by a four-area line method, and placing the inoculated culture medium in an incubator at 30 ℃ for culturing for 24-48 hours to test the purity and activation condition of the strain, so that the strain has good growth and reproduction capability;
(3) the primary expansion is carried out, and a Luria-Bertani culture medium (LB culture medium for short) is selected: 10g/L of Tryptone (Tryptone), 5g/L of Yeast extract (Yeast extract), 10g/L of sodium chloride (NaCl), regulating the pH of the culture medium to 7.2-7.4 by using 5% NaOH, supplementing distilled water to 1.0 g, sterilizing at 121 ℃ for 20min, taking out, placing into an inclined plane, inoculating under aseptic condition to respectively inoculate Bacillus altitudinis (GAU-00532), and a mother strain of Bacillus subtilis Bacillus subtilis (GAU-00533), placing in a constant temperature bacterial incubator, maintaining the temperature at 30 ℃, culturing for 48-72 h, and growing fungus fur plump, thus taking out for secondary strain production;
(4) expanding a secondary and shaking tables, selecting LB culture medium, inoculating 3 primary strains respectively under aseptic conditions, culturing at the temperature, placing on the shaking tables, and shake culturing for 12-24 h, wherein under normal conditions, the strain grows well, the bacterial liquid is viscous and has a small amount of foam;
(5) preparing a microorganism strain composition Bacillus altitudinis (GAU-00532) and a bacillus subtilis Bacillus subtilis subsp.Stercoris (GAU-00533) and a bacillus zhangzhouensis (GAU-00534) in a ratio of (0.8-1.2) to (1.6-2.5) to (1.0-1.8);
(6) detecting, wherein the effective viable count is determined by a spectrophotometry colorimetric method, and when OD is more than or equal to 0.5 and less than or equal to 1.0, the bacterial growth and reproduction capability is in a vigorous period;
(7) and (5) metering and packaging, wherein the error is 500+/-0.05 ml, and packaging by using an opaque plastic bottle.
(2) The results of the pig manure deodorization test showed that the removal rates of ammonia and hydrogen sulfide were 44% and 24.9%, respectively, after 72 hours (table 2).
TABLE 2 biological desulfurizing and deaminating deodorant pair H 2 S and NH 3 Is to (1) the treatment effect
Note that: CK means adding the same dose of sterile medium
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (3)
1. A desulfurizing and ammonia-removing microbial deodorant is characterized by comprising Bacillus altitudinis GAU-00532 and bacillus subtilis Bacillus subtilis subsp.Stercoris GAU-00533 and Bacillus zhangzhouensis GAU-00534, wherein the three bacillus are preserved in China center for type culture Collection, the preservation addresses are university of Wuhan in Wuhan, and the preservation numbers are CCTCC NO: M2021352, CCTCC NO: M2021350 and CCTCC NO: M2021351 respectively.
2. The desulfurizing and deaminizing microbial deodorant according to claim 1, wherein the ratio of Bacillus altitudinis GAU-00532 to Bacillus subtilis GAU-00533 and Bacilluszhangzhouensis GAU-00534 in the desulfurizing and deaminizing microbial deodorant is 0.8-1.2:1.6-2.5:1.0-1.8.
3. A method for producing the desulfurizing and ammonia-removing microbial deodorant according to claim 1 or 2, characterized by comprising: the Bacillus altitudinis GAU-00532 and Bacillus subtilis GAU-00533, bacilluszhangzhouensis GAU-00534 were cultured in LB medium at 30℃respectively, and then mixed.
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| CN101856505A (en) * | 2010-06-17 | 2010-10-13 | 宁津县瑞丰食品有限责任公司 | Biological deodorant for pig farm and preparation method thereof |
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| CN101856505A (en) * | 2010-06-17 | 2010-10-13 | 宁津县瑞丰食品有限责任公司 | Biological deodorant for pig farm and preparation method thereof |
| CN107325992A (en) * | 2017-08-31 | 2017-11-07 | 成都友益佳环保设备工程有限公司 | A kind of bacillus subtilis strain and its application |
| CN109609401A (en) * | 2018-12-18 | 2019-04-12 | 山东省科学院生态研究所 | A kind of complex microorganism deodorizing microorganism and the preparation method and application thereof |
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