CN114540240A - Microecological preparation for sewage treatment of farm, preparation method and application - Google Patents

Microecological preparation for sewage treatment of farm, preparation method and application Download PDF

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Publication number
CN114540240A
CN114540240A CN202210233525.4A CN202210233525A CN114540240A CN 114540240 A CN114540240 A CN 114540240A CN 202210233525 A CN202210233525 A CN 202210233525A CN 114540240 A CN114540240 A CN 114540240A
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China
Prior art keywords
bacillus subtilis
farm
preparation
sewage
treatment
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Inventor
佘德勇
何燕飞
刘琼
王一凡
汪雅
孙奎
韩伟峰
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Anhui Guonong Livestock And Poultry Breeding Technology Co ltd
West Anhui University
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Anhui Guonong Livestock And Poultry Breeding Technology Co ltd
West Anhui University
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Priority to CN202210233525.4A priority Critical patent/CN114540240A/en
Publication of CN114540240A publication Critical patent/CN114540240A/en
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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
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • 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
    • 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
    • C02F2101/30—Organic compounds
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
    • C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
    • 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
    • Y02W10/00—Technologies for wastewater treatment
    • Y02W10/10—Biological treatment of water, waste water, or sewage

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Microbiology (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Water Supply & Treatment (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Environmental & Geological Engineering (AREA)
  • Hydrology & Water Resources (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention discloses a microecological preparation for treating sewage in a farm, which comprises the active ingredient of bacillus subtilis and the dosage form of a bacterial liquid, a microbial agent or a carrier microbial agent. The invention also discloses a preparation method of the microecological preparation for the sewage treatment of the farm, which comprises the following steps: activating bacillus subtilis, inoculating the activated bacillus subtilis into an LB liquid culture medium, and culturing at 20-45 ℃ for 16-24 hours to obtain a bacterial liquid; centrifuging the bacterial liquid to obtain the microbial inoculum; and mixing the microbial inoculum with a carrier to obtain the carrier microbial inoculum. The bacillus subtilis can effectively treat the farm sewage with high COD concentration under the natural condition; the prepared bacterial liquid, bacterial agent and carrier bacterial agent can be applied to the treatment of sewage in a farm; and the treatment process is simple to operate, the effluent quality is good, and the treatment cost is low.

Description

Microecological preparation for sewage treatment of farm, preparation method and application
Technical Field
The invention relates to the technical field of sewage treatment, in particular to a microecological preparation for sewage treatment of a farm, and a preparation method and application thereof.
Background
The live pig is the main component of domestic animal breeding in China. In recent years, the annual culture quantity of live pig culture reaches 4200 thousands of pigs, 4000 thousands of high-quality pork products can be provided for residents in China every year, the consumption demand of meat products of people in China is guaranteed, the annual output value reaches more than 1 trillion, and the pork products account for 32.3% of animal husbandry economy. Therefore, the live pig breeding has important significance for realizing the rural economic development.
At present, the large-scale degree of pig breeding in China reaches 32.1%. The development bottleneck problem also appears in the pig industry development, and a large amount of waste is inevitably produced in the pig large-scale breeding, and particularly in a large-scale breeding farm which is raised in a highly concentrated manner, the annual production amount of the waste is very large.
COD (chemical Oxygen demand) is the amount of reducing substances to be oxidized in a water sample measured by a chemical method, and reflects the amount of reducing substances to be oxidized in sewage, and is an important organic pollution parameter which can be quickly measured in the research of river pollution and industrial sewage properties and the operation management of sewage treatment plants. With the rapid development of large-scale farms in recent years, dry-wet separation is realized in most farms, and excrement is returned to the field through composting, however, no good measure is provided for sewage treatment, and the discharge of the sewage causes pollution to the environment such as atmosphere, soil, water and the like to different degrees, so that the environment quality is seriously influenced.
Therefore, the development of relevant research aiming at the sewage treatment of the farm is of great significance.
Disclosure of Invention
In view of the above, the present invention aims to provide a microecological preparation for farm sewage treatment, and a preparation method and an application thereof, so as to solve the defects in the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
a microecological preparation for treating sewage in a farm comprises the active ingredient of bacillus subtilis, and the dosage form of the microecological preparation is bacterial liquid, a microbial agent or a carrier microbial agent.
Further, the Bacillus subtilis is Bacillus subtilis (Bacillus sp.) ACCC 62075.
The bacillus subtilis has the beneficial effects that: the bacillus subtilis can effectively treat the farm sewage with high COD concentration under natural conditions.
Further, the method for separating and screening the bacillus subtilis comprises the following steps:
(1) collecting a sample from sewage in a farm, inoculating the sewage sample on an LB solid culture medium by adopting a limiting dilution method, culturing at 37 ℃, selecting a single colony and purifying a strain;
(2) adding the strain into artificial sewage prepared in a laboratory according to the addition of 0.1 percent, and screening the strain;
(3) detecting the content of COD, total nitrogen and ammonia nitrogen in the artificial sewage, selecting a strain capable of efficiently treating the sewage, and performing sequencing identification.
The separation and screening method of the bacillus subtilis has the beneficial effects that: the method is efficient and rapid.
Further, the bacillus subtilis shows positive by gram staining, bacterial colonies are round when the bacillus subtilis is cultured on a flat plate, the surface of the bacillus subtilis is smooth and slightly convex, the edges of the bacillus subtilis are regular, the bacillus subtilis is red in color, and the bacillus subtilis is spherical after microscopic examination.
A preparation method of a microecological preparation for sewage treatment of a farm comprises the following steps: activating bacillus subtilis, inoculating the activated bacillus subtilis into an LB liquid culture medium, and culturing at 20-45 ℃ for 16-24h to obtain a bacterial liquid.
The bacterial liquid has the beneficial effects that: the bacterial liquid can be applied to the treatment of sewage in a farm.
A preparation method of a microecological preparation for sewage treatment of a farm comprises the following steps:
(1) activating bacillus subtilis for treating sewage in a farm, inoculating the activated bacillus subtilis into an LB liquid culture medium, and culturing at 20-45 ℃ for 16-24 hours to obtain a bacterial liquid;
(2) and centrifuging the bacterial liquid to obtain the microbial inoculum.
The microbial inoculum of the invention has the beneficial effects that: the microbial inoculum can be applied to the treatment of sewage in a farm.
A preparation method of a microecological preparation for sewage treatment of a farm comprises the following steps:
(1) activating bacillus subtilis for treating sewage in a farm, inoculating the activated bacillus subtilis into an LB liquid culture medium, and culturing at 20-45 ℃ for 16-24 hours to obtain a bacterial liquid;
(2) centrifuging the bacterial liquid to obtain a microbial inoculum;
(3) and mixing the microbial inoculum with a carrier to obtain the preparation.
The preparation of the invention has the beneficial effects that: the preparation can be applied to the treatment of sewage in a farm.
The invention also claims the application of the preparation or the preparation prepared by the preparation method in the sewage treatment of the farm.
Further, the treatment temperature is 20-35 ℃, and the treatment objects are COD, total nitrogen and ammonia nitrogen.
The application of the invention has the beneficial effects that: the treatment process is simple to operate, the effluent quality is good, and the treatment cost is low.
Drawings
FIG. 1 shows the effect of isolated strains (numbered FYHMWS101, FYHMWS102, FYHMWS103, FYHMWS104, and FYHMWS105, respectively) on the chemical oxygen demand in wastewater;
FIG. 2 shows the effect of isolated strains (numbered FYHMWS101, FYHMWS102, FYHMWS103, FYHMWS104, and FYHMWS105, respectively) on the total nitrogen content of wastewater;
FIG. 3 shows the effect of isolated strains (FYHMWS 101, FYHMWS102, FYHMWS103, FYHMWS104, and FYHMWS105, respectively) on the ammonia nitrogen content of wastewater;
FIG. 4 is a phylogenetic tree analysis of FYHMWS101 strain.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Bacillus subtilis (Bacillus sp.) ACCC 62075 in the following examples was collected by the Agricultural microorganism center of China Committee for Culture Collection of microorganisms (also called China Agricultural Culture Collection of China, ACCC for short, at the address of 010-82108636, China academy of Agricultural sciences, Ministry of south China, Ministry of agriculture, Ministry of Hokko, Hakko, Beijing), and the Collection date was 2022, 01-27 months, from which the strain was obtained by the public from the Collection center of the Agricultural microorganism Culture Collection.
Example 1 isolation of bacterial species
Diluting the sewage of a farm by 2 times, 4 times, 6 times and 8 times respectively, uniformly coating 50 mu L of diluent on a solid culture medium, culturing at 37 ℃ for 12h, picking a single colony to another new plate when the single colony grows on the plate, and purifying by a scribing method to obtain 5 strains which are named as FYHMWS101, FYHMWS102, FYHMWS103, FYHMWS104 and FYHMWS105 respectively.
Example 2 Sewage treatment Strain screening
The storage slant of the 5 selected microbial strains (FYHMWS 101, FYHMWS102, FYHMWS103, FYHMWS104, and FYHMWS105, respectively) was taken out, inoculated into 100mL of sterilized LB liquid medium, and cultured in a shaking incubator (37 ℃ C., 130rpm) for 24 hours. And (3) respectively detecting the bacterial liquids at the wavelength of 600nm by using an ultraviolet spectrophotometer, and adjusting the OD value to be 2.501 by using distilled water for later use.
Preparing simulated rural sewage with COD value of 300mg/L, autoclaving at 121 ℃ for 30min, and standing at room temperature until cooling for later use. Each 250mL triangular flask is filled with 100mL simulated rural sewage, 200 mu L of the prepared bacterial liquid is added into the simulated rural sewage, the mixture is continuously placed in an oscillation incubator (25 ℃, 250r/min) to be cultured for 12d, after standing for 30min, supernatant is taken to measure COD value, the COD removal rate is calculated by comparing the COD value with the COD value of the initial simulated rural sewage, and the result is shown in figure 1; the total nitrogen content is detected, and the result is shown in figure 2; the ammonia nitrogen content was measured and the results are shown in figure 3.
As can be seen from FIGS. 1-3, 5 microbial strains (numbered FYHMWS101, FYHMWS102, FYHMWS103, FYHMWS104 and FYHMWS105, respectively) screened by the present invention all significantly reduced COD, total nitrogen content and ammonia nitrogen content in wastewater.
Example 3 identification of FYHMWS101
FYHMWS101 strain was subjected to sequencing identification, and then subjected to evolutionary tree analysis based on the sequencing sequence, and the results are shown in FIG. 4.
As shown in FIG. 4, the FYHMWS101 strain was Bacillus subtilis.
Example 4
The preparation method of the microecological preparation for the sewage treatment of the farm comprises the following steps: activating Bacillus subtilis (Bacillus sp.) ACCC 62075, inoculating the activated Bacillus subtilis ACCC 62075 into an LB liquid culture medium, and culturing at 25 ℃ for 18h to obtain a bacterial liquid.
Example 5
The preparation method of the microecological preparation for the sewage treatment of the farm comprises the following steps:
(1) activating Bacillus subtilis (Bacillus sp.) ACCC 62075, inoculating the activated Bacillus subtilis ACCC 62075 into an LB liquid culture medium, and culturing at 25 ℃ for 18 hours to obtain a bacterial liquid;
(2) and centrifuging the bacterial liquid to obtain the microbial inoculum.
Example 6
The preparation method of the microecological preparation for the sewage treatment of the farm comprises the following steps:
(1) activating Bacillus subtilis (Bacillus sp.) ACCC 62075, inoculating the activated Bacillus subtilis ACCC 62075 into an LB liquid culture medium, and culturing at 25 ℃ for 18 hours to obtain a bacterial liquid;
(2) centrifuging the bacterial liquid to obtain a microbial inoculum;
(3) and mixing the microbial inoculum with a carrier to obtain the carrier microbial inoculum.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims (6)

1. A microecological preparation for treating sewage in a farm is characterized in that the active ingredient of the microecological preparation is bacillus subtilis, and the dosage form of the microecological preparation is bacterial liquid, a microbial inoculum or a carrier microbial inoculum.
2. The microecological preparation for farm wastewater treatment according to claim 1, wherein the Bacillus subtilis is Bacillus subtilis (Bacillus sp.) ACCC 62075.
3. A method for preparing a microecological formulation for sewage treatment in a farm according to claim 1 or 2, which comprises the steps of: activating bacillus subtilis, inoculating the activated bacillus subtilis into an LB liquid culture medium, and culturing for 16-24h at 20-45 ℃ to obtain the bacterial liquid.
4. A method for preparing a microecological preparation for sewage treatment of a farm according to claim 1 or 2, comprising the steps of:
(1) activating bacillus subtilis, inoculating the activated bacillus subtilis into an LB liquid culture medium, and culturing at the temperature of 20-45 ℃ for 16-24 hours to obtain a bacterial liquid;
(2) and (4) centrifuging the bacterial liquid to obtain the microbial inoculum.
5. A method for preparing a microecological preparation for sewage treatment of a farm according to claim 1 or 2, comprising the steps of:
(1) activating bacillus subtilis, inoculating the activated bacillus subtilis into an LB liquid culture medium, and culturing at the temperature of 20-45 ℃ for 16-24 hours to obtain a bacterial liquid;
(2) centrifuging the bacterial liquid to obtain a microbial inoculum;
(3) and mixing the microbial inoculum with a carrier to obtain the carrier microbial inoculum.
6. Use of the preparation according to claim 1 or 2 or the preparation prepared by the method according to any one of claims 3 to 5 in the treatment of sewage from a farm, wherein the treatment temperature is 20 to 35 ℃ and the treatment objects are COD, total nitrogen and ammonia nitrogen.
CN202210233525.4A 2022-03-10 2022-03-10 Microecological preparation for sewage treatment of farm, preparation method and application Pending CN114540240A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117070428A (en) * 2023-10-17 2023-11-17 北京大北农科技集团股份有限公司 Application of bacillus subtilis BS-22 strain in improving cultivation environment

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CN105779322A (en) * 2014-12-23 2016-07-20 北京大北农科技集团股份有限公司 Bacillus subtilis strain, microecological preparation, and applications thereof
CN105950507A (en) * 2016-05-24 2016-09-21 中国农业科学院农业环境与可持续发展研究所 Bacillus subtilis and bactericide as well as application of bactericide in treatment of livestock and poultry breeding waste water and treatment method
CN113943685A (en) * 2021-11-24 2022-01-18 上海中询生物科技有限公司 Culture method and application of special bacillus subtilis microbial inoculum for sewage

Patent Citations (4)

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Publication number Priority date Publication date Assignee Title
CN102703420A (en) * 2010-11-29 2012-10-03 王祥红 Microbial degradation preparation for municipal garbage and sewage and preparation method thereof
CN105779322A (en) * 2014-12-23 2016-07-20 北京大北农科技集团股份有限公司 Bacillus subtilis strain, microecological preparation, and applications thereof
CN105950507A (en) * 2016-05-24 2016-09-21 中国农业科学院农业环境与可持续发展研究所 Bacillus subtilis and bactericide as well as application of bactericide in treatment of livestock and poultry breeding waste water and treatment method
CN113943685A (en) * 2021-11-24 2022-01-18 上海中询生物科技有限公司 Culture method and application of special bacillus subtilis microbial inoculum for sewage

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117070428A (en) * 2023-10-17 2023-11-17 北京大北农科技集团股份有限公司 Application of bacillus subtilis BS-22 strain in improving cultivation environment
CN117070428B (en) * 2023-10-17 2024-02-27 北京大北农科技集团股份有限公司 Application of bacillus subtilis BS-22 strain in improving cultivation environment

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