CN117448206A - Bacillus pumilus and application thereof in culture water body regulation - Google Patents
Bacillus pumilus and application thereof in culture water body regulation Download PDFInfo
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 69
- 241000194103 Bacillus pumilus Species 0.000 title claims abstract description 44
- 230000015556 catabolic process Effects 0.000 claims abstract description 23
- 238000006731 degradation reaction Methods 0.000 claims abstract description 23
- 239000003242 anti bacterial agent Substances 0.000 claims abstract description 22
- 229940088710 antibiotic agent Drugs 0.000 claims abstract description 21
- 238000009629 microbiological culture Methods 0.000 claims abstract description 3
- 238000004321 preservation Methods 0.000 claims abstract 4
- 239000002068 microbial inoculum Substances 0.000 claims description 37
- 238000000855 fermentation Methods 0.000 claims description 23
- 230000004151 fermentation Effects 0.000 claims description 23
- 241000894006 Bacteria Species 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 18
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- 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
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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
- 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
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- 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
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- 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
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The invention discloses a Bacillus pumilus and application thereof in culture water regulation, wherein the preservation name of the Bacillus pumilus is Bacillus pumilus WR-1 (Bacillus pumilus), and the Bacillus pumilus is preserved in China general microbiological culture Collection center (CGMCC) at the 08-month 04 of 2023, and the preservation address is: beijing city, chaoyang district, north Chenxi lu 1, 3, accession number: CGMCC No.28097. The bacillus pumilus is derived from a culture water body treatment system of high-aquatic antibiotics, has strong capability of resisting aquatic antibiotics, has good COD degradation capability in a culture water body environment with high antibiotics, has strong stress resistance, has very obvious environmental adaptation capability, and can strengthen the original sewage treatment system.
Description
Technical Field
The invention relates to the field of aquaculture and microorganisms, in particular to bacillus pumilus and application thereof in culture water body regulation.
Background
At present, the pond culture mostly adopts an intensive and large-scale culture mode, along with overlarge stocking density and overlarge bait feeding amount, the residual feces and bait in the water body are increasingly accumulated, and feed and fishing drug additives are enriched, so that the water quality is deteriorated, and the content of aquatic antibiotics is too high. The main pollutants in the aquaculture tail water include ammonia nitrogen, nitrite, organic matters, phosphorus and fouling organisms, if the direct emission cannot be treated timely and effectively, the environment of the aquaculture water area is deteriorated, and the outbreak diseases such as fishes, shrimps and crabs and the like and even large-area death can be caused, so that the quality and the yield of the aquaculture products are directly reduced, even the human health is endangered, and the idea of green and healthy aquaculture is seriously violated.
The existing method for treating the water of the culture tail water mainly comprises a physical treatment technology, a chemical treatment technology and a biological treatment technology. The physical treatment technology is mainly a filtration method, a water mixing method, a foam separation method and a membrane separation method, and has the advantages of simple treatment mode and low cost, but the physical treatment technology can not reach the effluent quality standard under the normal condition and is difficult to control. The chemical treatment technology mainly comprises a chemical agent method and a chemical treatment technology method, and the chemical treatment technology has the advantages of good water outlet effect and high treatment efficiency, but the subsequent chemical agent treatment is difficult, and the secondary pollution problem can be caused. The biological treatment technology is mainly ecological treatment, biomembrane process and algae purifying process. The method mainly comprises a three-pond two-dam tail water treatment mode, an artificial wetland tail water treatment mode, a fishing rice co-cropping tail water treatment mode, a one-pond one-channel simple tail water treatment mode and other treatment processes, and a biological treatment method is applied to a plurality of treatment processes, so that auxiliary treatment of microbial agents is needed. Therefore, the separation and breeding of strains with high resistance to aquatic antibiotics and COD degradation capability are particularly important to be applied to biological treatment technology.
Disclosure of Invention
The invention aims to provide bacillus pumilus and application thereof in culture water body regulation, which can resist aquatic antibiotics, improve chemical oxygen demand removal efficiency in the culture water body and degrade total phosphorus and total nitrogen.
In order to achieve the above purpose, the technical scheme provided by the invention is as follows:
bacillus pumilus, which has a deposit name of Bacillus pumilus WR-1 (Bacillus pumilus), was deposited at China general microbiological culture Collection center (CGMCC), at a deposit address: beijing city, chaoyang district, north Chenxi lu 1, 3, accession number: CGMCC No.28097.
Further, the bacillus pumilus is a gram positive bacterium, and two different colony morphologies of semitransparent type and opaque type exist.
Further, the nucleotide sequence of the 16S rDNA of the bacillus pumilus is shown as SEQ ID NO. 1.
SEQ ID NO.1：
GATGGCTATACATGCAGTCGAGCGAACAGAAGGGAGCTTGCTCCCGGATGTTAGCGGCGGACGG
GTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAAACCGGAGCTAATACCGG
ATAGTTCCTTGAACCGCATGGTTCAAGGATGAAAGACGGTTTCGGCTGTCACTTACAGATGGACCCGC
GGCGCATTAGCTAGTTGGTGGGGTAATGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGT
GATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCC
GCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCT
GTTGTTAGGGAAGAACAAGTGCGAGAGTAACTGCTCGCACCTTGACGGTACCTAACCAGAAAGCCAC
GGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTA
AAGGGCTCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCCGGGGAGGGTCATTG
GAAACTGGGAAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGA
GATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGAGCGAAAGC
GTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGG
GGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGA
CTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAAC
GCGAAGAACCTTACCAGGTCTTGACATCCTCTGACAACCCTAGAGATAGGGCTTTCCCTTCGGGGACA
GAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGA
GCGCAACCCTTGATCTTAGTTGCCAGCATTTAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCG
GAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGG
ACAGAACAAAGGGCTGCGAGACCGCAAGGTTTAGCCAATCCCATAAATCTGTTCTCAGTTCGGATCG
CAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATA
CGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGG
TAACCTTTATGGAGCCAGCCGCCGAAGG
The invention also protects the microbial inoculum prepared by the bacillus pumilus.
Further, the microbial inoculum is a liquid microbial inoculum or a solid microbial inoculum.
And (3) carrying out high-density fermentation on the bacillus pumilus to obtain a fermentation product which is a liquid microbial inoculum.
As a preferred scheme, the preparation process of the liquid microbial inoculum comprises the steps of culturing bacillus pumilus to form bacterial colonies, inoculating the bacterial colonies into an LB liquid culture medium, carrying out shake culture at 170-230r/min, inoculating the bacterial colonies into another LB liquid culture medium, carrying out shake culture at 170-230r/min, and inoculating the bacterial colonies into a fermentation medium; aeration rate of fermentation culture is controlled to be 0.5-2:1, stopping fermentation until dissolved oxygen rises and pH drops; the culture temperature and time are 24-34 deg.C for 20-30 hr.
And (3) adsorbing, drying and crushing the fermentation product of the high-density fermentation to obtain the solid microbial inoculum.
The invention also protects the application of the bacillus pumilus or the microbial inoculum in the culture water body regulation, and the bacillus pumilus or the microbial inoculum is used as a culture water body regulator.
The bacillus pumilus can resist aquatic antibiotics in the environment of the aquaculture water body, and simultaneously degrade chemical oxygen demand, total phosphorus and total nitrogen in the aquaculture water body.
As a preferred embodiment, the degradation temperature is 24-30deg.C and the pH is 6.5-7.5.
Compared with the prior art, the invention has the beneficial effects that:
the bacillus pumilus is derived from a culture water body treatment system of high-aquatic antibiotics, has strong capability of resisting the aquatic antibiotics, has good COD degradation capability in a culture water body environment with the high antibiotics, has strong stress resistance and very remarkable environment adaptation capability, and can strengthen the original sewage treatment system by adding the bacillus pumilus into the sewage treatment system, so that more effective treatment effect is achieved.
The bacillus pumilus or the microbial inoculum thereof can be used as a culture water body regulator to improve the removal efficiency of chemical oxygen demand in the culture water body, has the capability of total phosphorus and total nitrogen in the aquaculture water body, has the advantages of simple operation, safety, environmental protection, strong adaptability, no secondary pollution and the like when being applied to the treatment of the culture tail water, can be widely applied to various culture water body treatment systems with high antibiotic concentration, and has wide application prospect and good environmental benefit.
Drawings
Fig. 1: phylogenetic tree of strain WR-1 constructed by Neighbor-joining method.
Fig. 2: figure of the effect of antibiotics on COD degradation rate in example 3.
Fig. 3: graph of the effect of temperature on COD degradation rate in example 3.
Fig. 4: the effect of pH on COD degradation rate in example 3 is shown.
Fig. 5: graph of COD over time in example 5.
Detailed Description
The above-described matters of the present invention will be further described in detail by way of examples, but it should not be construed that the scope of the above-described subject matter of the present invention is limited to the following examples, and all techniques realized based on the above-described matters of the present invention are within the scope of the present invention.
The experimental methods used in the examples below are conventional methods, and the reagents, methods and apparatus used are conventional in the art, unless otherwise indicated.
In describing the present invention, it should also be noted that:
bacillus pumilus WR-1 (Bacillus pumilus) in the embodiment of the invention is simply called a strain WR-1, and the microbial inoculum prepared by the Bacillus pumilus is simply called a WR-1 microbial inoculum.
The invention provides bacillus pumilus and application thereof in culture water regulation, wherein a water sample is collected from a culture water body with high antibiotic concentration, and is subjected to screening, extraction, analysis and purification, and gradient domestication, separation and purification in an inorganic salt culture medium to obtain an antibiotic-resistant COD degrading bacterium.
Through identification, the COD degrading bacteria are gram positive bacteria, two different colony forms of semitransparent bacteria and opaque bacteria exist, the two bacterial colonies are respectively passaged, the semitransparent bacteria can generate semitransparent bacteria and opaque bacteria, the semitransparent bacteria account for half of the semitransparent bacteria, the opaque bacteria only generate a little semitransparent bacteria in the passaging process, the semitransparent bacteria disappear when the opaque bacteria are transmitted to the 4 th generation, and the Bacillus pumilus WR-1 (Bacillus pumilus) is identified through 16S rDNA.
Further research experiments are carried out on the strain, and the strain can be applied to the treatment of aquaculture water, and can degrade COD, TP, TN and the like, especially has remarkable COD degradation effect, the initial COD concentration is 100mg/L, and the average degradation efficiency in 48 hours reaches more than 80%.
When the strain WR-1 is applied to the treatment of a culture water body, the strain WR-1 or a microbial inoculum thereof is directly added into a culture tail water treatment system, and aeration treatment is carried out to ensure that dissolved oxygen is more than 2mg/L, the pH of sewage is 5.0-7.5, and the temperature is preferably 25-35 ℃; the microbial inoculum can be a liquid microbial inoculum or a solid microbial inoculum.
As a further embodiment, the addition amount of the strain WR-1 during the application of the strain WR-1 is 10 according to the initial effective bacterial load of the system 5 -10 7 cfu/mL was added.
The invention further provides a WR-1 microbial inoculum prepared by the strain WR-1, and the preparation method comprises the following steps:
carrying out high-density fermentation on the bacterial strain WR-1 to obtain a liquid microbial inoculum thereof; and further adsorbing, drying and crushing the fermentation product to obtain the solid microbial inoculum.
The preparation process comprises the following steps:
firstly, inoculating a bacterial strain WR-1 on a test tube slant culture medium, and placing the bacterial strain WR-1 in a constant temperature incubator to culture for 20-30 hours at 24-34 ℃ until bacterial colonies grow on the culture medium;
step two, inoculating the strain into LB liquid medium (100 mL), shake culturing at 24-34 ℃ at about 170-230r/min for 20-30h, then inoculating 5% (v/v) into another LB liquid medium, culturing for 20-30h under the same condition, then inoculating into fermentation medium, fermenting, controlling the fermentation temperature at 24-34 ℃ and the ventilation rate at (0.5-2): 1, culturing for 20-30h, raising dissolved oxygen, lowering pH, stopping fermentation to obtain the liquid microbial inoculum.
Thirdly, further adsorbing, drying and crushing the fermentation product to obtain a solid microbial inoculum; wherein the adsorption, drying and pulverization are carried out according to the conventional technology in the field, and the detailed description is omitted.
Preferably, the liquid microbial agent has a viable count of 5×10 7 -1×10 10 cfu/mL; the viable count of the solid microbial inoculum is 2 multiplied by 10 10 cfu/g or more.
The invention is further described in detail below with reference to specific examples:
example 1: acquisition of WR-1 bacterium
The strain WR-1 is a strain which is obtained by sampling from a culture water body with high concentration of antibiotics and carrying out primary screening and secondary screening and is resistant to the bacteria of the antibiotics and capable of efficiently degrading COD, and the specific steps are as follows:
firstly, respectively taking water samples from three different places of a high salinity culture water body, respectively carrying out gradient dilution on an initial water sample in an ultra-clean workbench by using PBS buffer solution, fully shaking up, then sucking 50 mu L of dilution liquid, respectively coating the dilution liquid on a culture medium flat plate, and carrying out aerobic culture at 30 ℃ for 24-48 hours; after the colony is formed, picking a single colony, and carrying out plate streaking separation again to obtain a colony with single strain types; the isolated colonies were pelleted, stained, observed by microscopy for strain morphology and shape, plates were selected in spore form and stored at-80℃in 20% glycerol.
And (3) strain identification: the invention determines the 16S rDNA sequence of the strain; the primers for bacterial amplification adopt bacterial universal primers, and the forward primer is Eubac27F: (5-AGAGTTTGATC-CTGGCTCAG-3), reverse primer is Eubac1492R: (5-GGTTACCTTGTTACGACTT-3), mixing 25 μL 2 xMix Taq with 2 μL of each of forward and reverse primers and 2 μL of bacterial liquid, adding water to 50 μL system, and performing PCR amplification; the PCR reaction procedure is 94 ℃ pre-denaturation for 4min; denaturation at 94℃for 30s; renaturation at 55 ℃ for 1min; extending at 72 ℃ for 1.5min for 30 cycles; extending at 72 ℃ for 10min; detecting PCR products by using 1% agarose gel electrophoresis, sending to a sequencing company for sequencing, and searching and comparing the strain 16S rDNA sequence results in NCBI database by using a BLAST method; the 16S rDNA sequences of the strain WR-1 and the similar strains are subjected to multi-sequence alignment by means of Mega (version 7.0) software, and a phylogenetic tree is constructed by adopting a Neighbor-joining method. As shown by the phylogenetic tree results in FIG. 1, the strain is Bacillus pumilus.
Example 2: preparation of WR-1 microbial inoculum
The preparation method of the WR-1 microbial inoculum in the embodiment comprises the following steps:
firstly, inoculating the purified strain WR-1 on a test tube slant culture medium, and placing the strain WR-1 in a constant temperature incubator to culture at 24-34 ℃ until bacterial colonies grow on the culture medium;
secondly, transferring the strain into LB liquid medium (100 mL), shake culturing at 24-34 ℃ and 180r/min for 24h, inoculating 5% (v/v) to another LB liquid medium, culturing again under the same condition for 24h, inoculating to fermentation medium, fermenting, controlling the fermentation temperature at 24-34 ℃ and ventilation volume at 1: about 1, culturing for 24 hours, raising dissolved oxygen, lowering pH, and stopping fermentation to obtain the WR-1 microbial inoculum.
The LB culture medium is prepared from 10.0g of peptone, 5.0g of yeast powder and 10.0g of sodium chloride by adding water to 1.0L, adjusting the pH value to 7.0, and sterilizing at 121 ℃ for 20 min.
Example 3: COD degradation characteristic experiment
WR-1 microbial inoculum was prepared as in example 2, with culture temperatures of 25.+ -. 1 ℃.
(1) Detection of maximum degradation capability:
preparing water samples with the concentration of 0.01, 0.05, 0.1 and 0.5mg/L of aquatic antibiotics, preparing three COD concentration gradients of 50, 100 and 500mg/L of each water sample with the concentration of the antibiotics, and adding 5mL of effective viable bacteria with the number of about 1.2 multiplied by 10 8 cfu/mL of the microbial inoculum is subjected to shake culture at 25 ℃ at 160r/min, sampling is carried out every 48h, the COD residual concentration is detected, the detection result shows that when the aquatic antibiotic concentration is below 0.1mg/L, the initial COD concentration is 100mg/L, and the average degradation rate of 48h reaches more than 80%, and the specific result is shown in figure 2.
(2) Impact test of culture conditions:
adopting 6 groups of water samples with the antibiotic concentration of 0.1mg/L and the COD of 100mg/L, setting different culture temperatures (respectively setting 20, 25, 28, 30, 35 and 40 ℃) for each group, and culturing for 48 hours at the rotating speed of 160 r/min;
10 groups of water samples with the antibiotic concentration of 0.1mg/L and the COD of 100mg/L are adopted, and each group of water samples is set with different pH (4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8 and 8.5) conditions, and is cultured for 48 hours at the temperature of 25 ℃ and the rotating speed of 160 r/min;
the culture solution after the culture is sampled to detect COD degradation condition, as shown in figures 3 and 4, the strain has wider application range, different degradation capacities under different temperature and pH conditions, and the optimal action temperature is 25-30 ℃ and the optimal action pH is 6.5-7.5.
Example 4: TP and TN degradation experiment
Setting 1 group of water samples with the antibiotic concentration of 0.1mg/L and the TP of 2.5mg/L, 1 group of water samples with the antibiotic concentration of 0.1mg/L and the TN of 25mg/L, picking a bacterial strain WR-1 single colony, inoculating the bacterial strain WR-1 single colony into 10mL of LB liquid medium, and inoculating the bacterial strain WR-1 single colony into the 2 groups of culture mediums respectively with the inoculation amount of 2% in the next day.
The culture conditions are as follows: culturing in a shaking table with rotation speed of 160rpm at 25 ℃ for 48 hours, detecting the residual concentrations of TP and TN, and calculating the degradation rate.
Finally, the following steps are: the degradation rates of the strain WR-1 on TP and TN in 48 hours are 34.6 percent and 46.7 percent respectively. The strain WR-1 has certain degradation capacity to TP and TN, and has wide application prospect.
Example 5: application 1 in aquaculture water treatment
A WR-1 microbial inoculum was prepared in the same manner as in example 2 at a culture temperature of 25.+ -. 1 ℃ to give a viable count of 1.2X10 8 cfu/mL。
The source of the treated water is a culture water body treatment system, a laboratory simulates aeration treatment of a biochemical system, the treatment capacity is 5L, the addition amount of a bacterial strain WR-1 is 0.1% (v/v), the aeration treatment enables dissolved oxygen to be more than 2mg/L, the pH of sewage to be 6.0-8.0, antibiotics to be about 0.02mg/L, COD to be about 80mg/L, TN to be about 20mg/L, TP to be 2.5mg/L, the initial COD concentration in the artificial addition system is 100mg/L, water inlet and outlet are carried out for 48h for 1d, the COD concentration in the water inlet and outlet maintenance system is 100mg/L each time, the COD content of water is sampled and detected, and the tracking experiment result is shown in Table 1, and the specific result is shown in figure 5.
TABLE 1COD degradation experiment results
Example 6: application 2 in high antibiotic aquaculture water treatment
Preparation of WR-1 inoculant according to the method of example 2, culture temperature of 27+ -1deg.CThe viable count of the microbial inoculum is 3.4X10 8 cfu/mL。
The source of the treated water is a pond culture tail water treatment system, a laboratory simulates aeration treatment of a biochemical system, the treatment capacity is 5L, the addition amount of WR-1 bacteria is 0.2% (v/v), the aeration treatment enables dissolved oxygen to be more than 2mg/L, the pH of sewage is 6.0-8.0, antibiotics are about 0.05mg/L, the initial concentration of antibiotics in the addition system is considered to be 0.1mg/L, COD is about 80mg/L, TN is about 20mg/L, TP is about 2.5mg/L, water is fed and discharged for 1 hour, water is fed and discharged for 1L each time, the content of COD, TP and TN is sampled and detected, the COD degradation rate reaches >82%, the TP degradation rate reaches >39%, and the TN degradation rate reaches >48%.
In conclusion, the strain WR-1 is derived from a culture tail water treatment system of high-aquatic antibiotics, indigenous microorganisms capable of degrading COD are screened from the strain, and the strain has the characteristics of simplicity and high efficiency in application, is strong in tolerance, can resist the concentration of the aquatic antibiotics to be 0.1mg/L or higher, has wide application range on conditions such as temperature, pH and the like, is not easy to be impacted by external environment in the application process, and has the capability of degrading TP and TN in a culture water body; the water treatment agent is added into a culture tail water treatment system with high antibiotic concentration, can effectively remove COD, has a certain removal effect on TP and TN, has good application value in actual production, and has wide application prospect.
The present invention is not limited to the preferred embodiments, and any simple modification, equivalent replacement, and improvement made to the above embodiments by those skilled in the art without departing from the technical scope of the present invention, will fall within the scope of the present invention.
Claims (10)
1. A bacillus pumilus, characterized in that: the preservation name of the Bacillus pumilus is Bacillus pumilus WR-1 (Bacillus pumilus), and the Bacillus pumilus is preserved in China general microbiological culture collection center (CGMCC) at the 08 th month 04 of 2023, and the preservation address is: beijing city, chaoyang district, north Chenxi lu 1, 3, accession number: cgmccno.28097.
2. The bacillus pumilus according to claim 1, wherein: the bacillus pumilus is a gram-positive bacterium, and has two different colony morphologies of semitransparent type and opaque type.
3. The bacillus pumilus according to claim 1, wherein: the nucleotide sequence of the 16S rDNA of the bacillus pumilus is shown as SEQ ID NO. 1.
4. The microbial inoculum prepared by the bacillus pumilus as set forth in claim 1, which is characterized in that: the microbial inoculum is liquid microbial inoculum or solid microbial inoculum.
5. The microbial agent of claim 4, wherein: and (3) carrying out high-density fermentation on the bacillus pumilus to obtain a fermentation product which is a liquid microbial inoculum.
6. The microbial agent of claim 5, wherein: the preparation process of the liquid microbial inoculum comprises the steps of culturing bacillus pumilus to form colonies, inoculating the colonies into an LB liquid culture medium, carrying out shake culture at 170-230r/min, inoculating the colonies into another LB liquid culture medium, and carrying out shake culture at 170-230r/min and then inoculating the colonies into a fermentation culture medium; aeration rate of fermentation culture is controlled to be 0.5-2:1, stopping fermentation until dissolved oxygen rises and pH drops; the culture temperature and time are 24-34 deg.C for 20-30 hr.
7. The microbial agent of claim 5, wherein: and (3) adsorbing, drying and crushing the fermentation product of the high-density fermentation to obtain the solid microbial inoculum.
8. Use of the bacillus pumilus of claim 1 or the microbial inoculum of claim 5 for the regulation of aquaculture water, characterized in that: as a culture water body regulator.
9. The use according to claim 8, characterized in that: the bacillus pumilus can resist aquatic antibiotics in the environment of the aquaculture water body, and simultaneously degrade chemical oxygen demand, total phosphorus and total nitrogen in the aquaculture water body.
10. The use according to claim 9, characterized in that: the degradation temperature is 25-30deg.C, and the pH is 6.5-7.5.
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