Application of pseudomonas PSC001 in degradation of aflatoxin B1 and promotion of rumen fermentation

Abstract

The invention relates to application of pseudomonas PSC001 in degradation of aflatoxin B1 and promotion of rumen fermentation, and aims to solve the problems of pollution of aflatoxin B1 in feed and poor ruminant rumen fermentation function at the same time. The invention discloses the new application of the pseudomonas PSC001 with the preservation number of CCTCC M 2023814 for the first time, namely degradation of aflatoxin B1 and promotion of rumen fermentation. The key technical means is as follows: the strain or the microbial inoculum thereof is applied to an AFB1 pollution system or daily ration of ruminants, and can realize efficient detoxification and growth promotion under specific conditions (such as 30-42 DEG C and pH 5.0-8.0) for 24-60 hours. The highest degradation rate of the strain to AFB1 can reach 65.20%, the yield of volatile fatty acid in rumen can be remarkably increased, a microflora is optimized, and the strain has the dual effects of efficient detoxification and rumen health improvement, and is high in safety and wide in application prospect.

Description

Technical Field

[0001] This invention relates to the field of microbial application technology, specifically to a strain of Pseudomonas aeruginosa with accession number CCTCC M 2023814. Pseudomonas sp .) New uses for PSC001, particularly its application in the degradation of aflatoxin B1 (AFB1) and in promoting rumen fermentation in ruminants. Background Technology

[0002] Aflatoxin B1 (AFB1) and zearalenone (ZEN) are two of the most dangerous mycotoxins in animal feed and agricultural products. Aflatoxin B1 is mainly produced by Aspergillus flavus (…). Aspergillus flavus ) and parasitic aspergillus ( Aspergillus parasiticus Aflatoxin B1 is produced by bacteria and possesses strong carcinogenicity, mutagenicity, and hepatotoxicity, seriously endangering the health of livestock and poultry and threatening human food safety through the food chain. Currently, detoxification methods for aflatoxin B1 include physical adsorption, chemical treatment, and microbial degradation. Physical and chemical methods suffer from drawbacks such as low efficiency, significant nutrient loss, and potential introduction of secondary pollution. Microbial methods have become a research hotspot due to their high efficiency, specificity, and environmental friendliness; however, currently reported aflatoxin B1 degrading strains have issues such as low degradation efficiency and questionable safety (e.g., they may be pathogenic).

[0003] On the other hand, the production efficiency of ruminants is highly dependent on their rumen fermentation function. A healthy rumen microbiota can efficiently break down cellulose, producing nutrients such as volatile fatty acids (VFAs). In actual production, factors such as mycotoxin contamination and improper diet structure can disrupt the rumen microecological balance, leading to weakened fermentation function and decreased feed conversion ratio. Therefore, developing safe and efficient biological agents that can simultaneously address mycotoxin contamination and improve rumen health is of great significance to the development of modern animal husbandry.

[0004] Patent document CN118389312A discloses the application of *Pseudomonas* PSC001 (accession number: CCTCC M 2023814) in the degradation of zearalenone (ZEN). However, whether this strain possesses the ability to degrade aflatoxin B1, and whether it can positively affect the rumen fermentation function of ruminants, remains unknown. Exploring novel functional applications of the same strain is of great significance for expanding its application value and reducing research and development costs.

[0005] The information disclosed in this background section is intended only to enhance the understanding of the background technology of this disclosure and should not be construed as an admission or in any way implying that the information constitutes prior art known to those skilled in the art. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a new use for Pseudomonas aeruginosa PSC001 with accession number CCTCC M 2023814. This invention is the first to discover that this strain can not only efficiently degrade aflatoxin B1 (AFB1), but also significantly promote rumen fermentation in ruminants, exhibiting a dual effect of "detoxification" and "growth promotion," providing a safe, efficient, and multifunctional microbial resource for solving feed safety and breeding efficiency issues.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: Firstly, the *Pseudomonas* strain with accession number CCTCC M 2023814 (…) Pseudomonas sp .) PSC001 is applied in at least one of the following (1) to (6): (1) Degrading aflatoxin B1 or preparing preparations that degrade aflatoxin B1; (2) Promote rumen fermentation in ruminants or prepare preparations that promote rumen fermentation in ruminants; (3) Increase the production of propionic acid, butyric acid, valeric acid, isovaleric acid, and total volatile fatty acids in rumen fluid; (4) Improve the rumen microbiota; (5) Increase the relative abundance of fiber-degrading bacteria genera; (6) with Cu 2+ Combined use to enhance the degradation effect of aflatoxin B1.

[0008] In some embodiments of this application, the degradation treatment conditions are: culture temperature of 30 ℃ to 42 ℃, pH value of 5.0 to 8.0, and treatment time of 24 h to 60 h.

[0009] In some embodiments of this application, the concentration of *Pseudomonas* added to the rumen fermentation broth is 1 × 10⁻⁶. 9 CFU / mL ~ 8×10 9 CFU / mL.

[0010] Secondly, a bacterial agent is provided, the active ingredient of which is *Pseudomonas* strain with accession number CCTCC M 2023814. Pseudomonas sp PSC001, the microbial agent has the functions of degrading aflatoxin B1 and promoting rumen fermentation in ruminants.

[0011] In some embodiments of this application, the bacterial agent is a liquid preparation or a lyophilized powder preparation.

[0012] This invention provides a strain of Pseudomonas aeruginosa with accession number CCTCC M 2023814. Pseudomonas sp Application of PSC001 in the degradation of aflatoxin B1.

[0013] Thirdly, the present invention provides a strain of Pseudomonas aeruginosa with accession number CCTCC M 2023814 ( Pseudomonas sp Application of PSC001 in the preparation of formulations for degrading aflatoxin B1.

[0014] Fourthly, the present invention provides a *Pseudomonas* strain with accession number CCTCC M 2023814 (…). Pseudomonas sp Application of PSC001 in promoting rumen fermentation in ruminants.

[0015] Fifthly, the present invention provides a *Pseudomonas* strain with accession number CCTCC M 2023814 (…). Pseudomonas sp Application of PSC001 in the preparation of formulations for promoting rumen fermentation in ruminants.

[0016] Preferably, the degradation conditions for the application of the aflatoxin B1 degradation include: culture temperature of 30℃~42℃, pH 5.0~8.0, and treatment time of 24 h~60 h.

[0017] More preferably, Cu is added to the reaction system. 2+ (e.g., CuSO4) can significantly enhance its degradation effect on AFB1, with a degradation rate of over 65%.

[0018] Preferably, the promotion of rumen fermentation in ruminants is manifested in significantly increasing the production of propionic acid, butyric acid, valeric acid, isovaleric acid, and total volatile fatty acids in rumen fluid, optimizing the rumen microbiota, and increasing the relative abundance of fiber-degrading bacteria.

[0019] Preferably, the amount of microbial agent added to promote rumen fermentation is 1×10⁻⁶. 9 CFU / mL ~ 8×10 9 CFU / mL rumen fermentation broth

[0020] Sixthly, the present invention provides a microbial agent, the active ingredient of which is *Pseudomonas* strain with accession number CCTCC M 2023814. Pseudomonas sp PSC001, the microbial agent has the functions of degrading aflatoxin B1 and promoting rumen fermentation.

[0021] Preferably, the bacterial agent is prepared by inoculating Pseudomonas PSC001 into a culture medium and culturing it at 10℃ to 42℃ until the bacterial concentration reaches 1×10⁻⁶. 7 CFU / mL ~ 1×10 9 It can be made from CFU / mL; or further processed into lyophilized powder formulations.

[0022] One or more technical solutions provided in the embodiments of this application have at least the following technical effects or advantages: 1. Functional Innovation: For the first time, a novel function of the known strain PSC001 (CCTCC M 2023814) in degrading AFB1 and promoting rumen fermentation has been revealed, expanding the application scope of this strain and realizing "one strain with multiple functions".

[0023] 2. High efficiency and safety: This strain originates from the rumen of healthy cattle and is a non-pathogenic, resident bacterium with high safety. It exhibits high degradation efficiency against AFB1 and maintains good activity in the acidic environment of the rumen, making it highly suitable for use in ruminant production.

[0024] 3. Synergistic Effect: This invention is the first to combine "mycotoxin degradation" with "rumen function regulation." Applying this microbial agent can improve animal digestive function and increase feed utilization while simultaneously detoxifying, ultimately achieving the goal of promoting animal growth and enhancing the economic benefits of animal husbandry.

[0025] 4. Broad application prospects: This microbial agent can be used to prepare feed additives, fermentation agents, etc., to purify feed raw materials contaminated with aflatoxin B1, and to improve the health of ruminants. It has huge market application potential and industrialization value. Attached Figure Description

[0026] Figure 1 This is a graph showing the relationship between the efficiency of strain PSC001 in degrading aflatoxin B1 and the reaction time in one embodiment of this disclosure.

[0027] Figure 2 This is a graph showing the relationship between the efficiency of strain PSC001 in degrading aflatoxin B1 and the reaction temperature in one embodiment of this disclosure.

[0028] Figure 3 This is a graph showing the relationship between the efficiency of strain PSC001 in degrading aflatoxin B1 and the reaction pH in one embodiment of this disclosure.

[0029] Figure 4 This diagram illustrates the effect of different metal ions on the degradation of aflatoxin B1 by strain PSC001 in one embodiment of this disclosure.

[0030] Figure 5 This is a graph showing the linear relationship between predicted and actual values ​​in one embodiment of this disclosure.

[0031] Figure 6 This is a functional abundance analysis diagram of FAPROTAX in two embodiments of this disclosure. Detailed Implementation

[0032] To better understand the technical solution of this application, the above technical solution will be described in detail below with reference to the accompanying drawings and specific embodiments.

[0033] Strain information: The strain involved in this invention is *Pseudomonas* (…). Pseudomonas sp PSC001, deposited at the China Center for Type Culture Collection (CCTCC), accession number CCTCC M 2023814, on May 24, 2023.

[0034] Culture media and reagents involved in the examples: 1. The preparation methods of LB liquid culture medium, MS liquid culture medium, and zearalenone stock solution are the same as those in the background art, cited in patent CN118389312A.

[0035] 2. Aflatoxin B1 stock solution: 10 mg of aflatoxin B1 standard was dissolved in 10 mL of chromatographic grade methanol to a concentration of 1 mg / mL. The solution was sterilized by filtration through a 0.22 μm filter membrane, aliquoted, and stored at -20°C protected from light.

[0036] 3. Aflatoxin B1 ELISA test kit: purchased from Beijing Yisenbao Biotechnology Co., Ltd.

[0037] 4. Rumen fluid: Collected from healthy beef cattle, filtered through four layers of gauze, then filled with CO2 and kept warm at 39℃ for later use.

[0038] Example 1: Verification of the ability of strain PSC001 to degrade aflatoxin B1 1. Relationship between the ability of Pseudomonas PSC001 to degrade aflatoxin B1 and reaction time. Single colonies cultured on blood agar plates were inoculated into LB liquid medium and incubated at 160 rpm and 37°C until OD500 was achieved. 600 Remove when the value is approximately 0.6 (logarithmic growth phase of bacteria).

[0039] Experimental group: 975 μL of bacterial culture was added to a 5 mL sterile EP tube, followed by 25 μL of aflatoxin B1 working solution (10 μg / mL), to bring the initial aflatoxin B1 concentration to 0.25 μg / mL. The tube was then incubated at 37℃ and 160 r / min for 12 h, 24 h, 36 h, 48 h, and 60 h, respectively, before the degradation rate was determined by ELISA.

[0040] The results are as follows: the effect of different degradation times on the degradation of aflatoxin B1 by Pseudomonas aeruginosa is as follows: Figure 1As shown, the degradation of aflatoxin B1 occurred rapidly in the first 24 hours, with a degradation rate of up to 52.14%. At this time, Pseudomonas may be in the logarithmic phase with high degradation activity. As time went on, the degradation of aflatoxin B1 by Pseudomonas reached its highest value of 54.37% at 48 hours. Finally, the degradation rate decreased to 50.17% at 60 hours. This may be because the long-term culture caused some Pseudomonas cells to die and lyse, or because substances produced by the degradation of aflatoxin B1 destroyed its activity.

[0041] 2. Relationship between the ability of Pseudomonas PSC001 to degrade aflatoxin B1 and reaction temperature. Single colonies cultured on blood agar plates were inoculated into LB liquid medium and incubated at 160 rpm and 37°C until OD500 was achieved. 600 Remove when the value is approximately 0.6 (logarithmic growth phase of bacteria).

[0042] Experimental group: 975 μL of bacterial culture was added to a 5 mL sterile EP tube, followed by 25 μL of aflatoxin B1 working solution (10 μg / mL), to bring the initial aflatoxin B1 concentration to 0.25 μg / mL. The tube was then incubated at 30℃, 34℃, 38℃, 42℃, and 46℃ at 160 r / min for 48 h. The degradation rate was then determined using ELISA.

[0043] The results are as follows: The effect of different degradation temperatures on the degradation of aflatoxin B1 by Pseudomonas aeruginosa is as follows: Figure 2 As shown, Pseudomonas aflatoxin B1 was degraded at temperatures ranging from 31°C to 43°C, with a degradation rate exceeding 52%. The degradation rate reached its highest point at 58.66% between 34°C and 37°C. This may be because Pseudomonas aflatoxin B1 is more suitable for growth at temperatures between 34°C and 37°C. At this temperature, Pseudomonas aflatoxin B1 has a vigorous metabolism, sufficient energy supply, and higher activity of the secreted degrading enzymes.

[0044] 3. Relationship between the ability of Pseudomonas PSC001 to degrade aflatoxin B1 and the reaction pH. Single colonies cultured on blood agar plates were inoculated into LB liquid medium and incubated at 160 rpm and 37°C until OD500 was achieved. 600 Remove when the value is approximately 0.6 (logarithmic growth phase of bacteria).

[0045] Experimental group: 975 μL of bacterial culture was added to a 5 mL sterile EP tube, followed by 25 μL of aflatoxin B1 working solution (10 μg / mL), to bring the initial aflatoxin B1 concentration to 0.25 μg / mL. The tube was then incubated at 37℃ and 160 rpm at pH 4.0, 5.0, 6.0, 7.0, 8.0, and 9.0 (adjusted with 1 mol / L hydrochloric acid or sodium hydroxide solution) for 48 h. The degradation rate was then determined using ELISA.

[0046] The results are as follows: The effect of different pH values ​​on the degradation of aflatoxin B1 by Pseudomonas aeruginosa is as follows: Figure 3 As shown, the degradation rate of aflatoxin B1 by Pseudomonas aeruginosa reached 58.98% at pH = 7. The degradation rate of aflatoxin B1 decreased under both acidic and alkaline conditions, and the decrease was more significant under alkaline conditions than under acidic conditions. This may be because Pseudomonas aeruginosa was isolated from the bovine rumen, where the environment is acidic. At the same time, under alkaline conditions, various intracellular enzymes that degrade aflatoxin B1 may be more easily degraded and lose their activity, thus reducing the degradation rate.

[0047] 4. The relationship between the ability of Pseudomonas PSC001 to degrade aflatoxin B1 and metal ions. Single colonies cultured on blood agar plates were inoculated into LB liquid medium and incubated at 160 rpm and 37°C until OD500 was achieved. 600 Remove when the value is approximately 0.6 (logarithmic growth phase of bacteria).

[0048] Experimental group: 975 μL of bacterial culture was added to a 5 mL sterile EP tube, followed by 25 μL of aflatoxin B1 working solution (10 μg / mL), to bring the initial aflatoxin B1 concentration to 0.25 μg / mL. The tube was then incubated at 37℃ and 160 r / min for 48 h in FeCl3, MgCl2, ZnSO4, CuSO4, and MnCl2 (adjusted with 1 mol / L hydrochloric acid or sodium hydroxide solution), respectively. The degradation rate was then determined by ELISA.

[0049] The results are as follows: The effects of different metal ions on Pseudomonas aeruginosa on aflatoxin B1 are as follows: Figure 4 As shown, different metal ions all have a promoting effect on the degradation of aflatoxin B1 by Pseudomonas, but Mn 2+ Fe 3+ The promoting effect was not significant, while Cu 2+ The most effective degradation rate reached 65.20%.

[0050] 5. Response surface methodology for the degradation of aflatoxin B1 by Pseudomonas PSC001 Based on previous in vitro single-factor experimental data, degradation temperature (°C), pH, and degradation time (h) were selected as three key independent variables (Xi). The degradation rate of aflatoxin B1 was set as the response value (Y). Using the Box-Behnken design pattern in response surface methodology, a three-factor, three-level experimental system was constructed using the Design Expert 13.0 program. The experimental design and results are shown in Table 1.

[0051] Table 1. Response surface methodology experimental design and results .

[0052] Import the data from Table 1 into Design Expert 13.0 for fitting analysis to obtain a binary multiple regression equation: Y=65.20+0.24A-3.81B-1.60C-1.20AB-0.65AC-1.33BC-3.35A 2 -8.48B 2 -4.73C 2 .

[0053] The significance test and analysis of variance of the regression equation are shown in Table 2. The regression equation model has a P value of 0.0051, which indicates that the model is significant. The lack-of-fit term has a P value of 0.4641, which indicates that the regression equation model has a good fit and small experimental error.

[0054] Based on the data analysis in Table 2, we can see that the linear term B and the quadratic term A... 2 B 2 C 2 The degradation rate of aflatoxin B1 was significantly reduced in all cases. P <0.05), while the degradation rates of the first-order terms A, C, AB, AC, and BC were not significant ( P >0.05), analysis of the phenomenon shows that time has a greater impact on the degradation rate of aflatoxin B1 by Pseudomonas, while temperature and pH have a smaller impact.

[0055] Table 2. Significance and ANOVA of Response Surface Regression Equations .

[0056] The coefficient of determination R of the regression equation constructed in this experiment 2 A value of 0.9159 indicates that the model explains 91.59% of the dependent variable. Figure 5 As shown, the regression model's predicted curve and the actual measured value are in good agreement, and the linear correlation test between the two passes the significance level ( ). P <0.05).

[0057] Response surface methodology (RSM) optimization experiments, analyzed and predicted using Design Expert 13.0, revealed that the optimal conditions for the degradation of aflatoxin B1 by *Pseudomonas* were a degradation time of 45.36 h, a degradation temperature of 34.27 ℃, and a degradation pH of 6.86, at which point the degradation rate reached 65.75%. Using an actual degradation time of 45 h, a degradation temperature of 34 ℃, and a degradation pH of 6.80, the experiment was repeated three times, yielding an actual degradation rate of 64.86%, which was close to the predicted result, indicating that the regression equation model is relatively realistic and reliable.

[0058] Example 2: Effects of strain PSC001 on in vitro rumen fermentation 1. Effects on rumen fermentation parameters and volatile fatty acid (VFA) yield Experiments were conducted using a dual-outflow continuous rumen culture simulation system, divided into three cycles, each lasting seven days. The system, through an intelligent control system, used a stepper motor to agitate the contents of the fermenter at 25 r / min. Buffer solution was introduced into the fermenter via a pressure pump at a rate of 6% per hour. Simultaneously, the overflow bottle's tubing was monitored to ensure timely collection of overflow liquid and undegraded solid fermentation substrate from each fermenter, ending in the overflow bottle.

[0059] The experiment consisted of two treatment groups: a control group (CON) and a Pseudomonas group (PS). The first three days were an acclimatization period, during which the control group was fed 20 g of unadulterated feed at 8:00 AM and 8:00 PM daily. After the acclimatization period, the control group received normal feed; the PS group received 5 mL of 4×10⁻⁶ Pseudomonas aeruginosa supplemented with TMR at 8:00 AM. 8 The fermentation tank was filled with Pseudomonas aeruginosa at CFU / mL, and fed only with standard TMR feed at 8:00 PM. Samples were taken from the fermenter on day 7 of each experimental period for pH, ammonia nitrogen, microbial protein, and volatile fatty acids.

[0060] The results of pH, NH3-N, and MCP measurements (Table 3) showed no significant changes in pH among the groups, while the NH3-N content in the PS group was significantly lower. Changes in volatile fatty acids in the artificial rumen (Table 4) showed that, compared to the CON group, the PS group had increased total volatile fatty acids and acetic acid, significantly or extremely significantly increased concentrations of propionic acid, valeric acid, and butyric acid, and extremely significantly decreased ethyl-propionic acid ratio and isovaleric acid concentration.

[0061] Table 3. Fermentation parameters in artificial rumen Data from the same peer group with no letter on the top label or the same lowercase letter indicates that the difference is not significant. P >0.05), different lowercase letters indicate significant differences ( P<0.05), different uppercase letters indicate extremely significant differences ( P <0.01), the same as in the table below.

[0062] Table 4. VFA content in artificial rumen .

[0063] 2. The impact of Pseudomonas on the rumen microbiota The experiment was conducted using a dual-outflow continuous culture rumen simulation system, consisting of three cycles, each lasting seven days. The system employed an intelligent control system with a stepper motor to agitate the fermenter contents at 25 r / min. Buffer solution was pumped into the fermenter at a rate of 6% per hour using a pressure pump. Simultaneously, the overflow bottle's tubing was monitored to ensure timely collection of overflow liquid and undegraded solid fermentation substrate from each fermenter, ending in the overflow bottle.

[0064] The experiment consisted of two treatment groups: a control group (CON) and a Pseudomonas group (PS). The first three days were an acclimatization period, during which the control group was fed 20 g of unadulterated feed at 8:00 AM and 8:00 PM daily. After the acclimatization period, the control group received normal feed; the PS group received 5 mL of 4×10⁻⁶ Pseudomonas aeruginosa supplemented with TMR at 8:00 AM. 8 CFU / mL of Pseudomonas aeruginosa culture was administered at 8:00 PM, fed only with standard TMR feed. Samples were taken from the fermenter on day 7 of each experimental period. DNA was extracted from the rumen fermentation broth on the last day of culture using a magnetic bead-based fecal DNA extraction kit (B618763-0100). The qualified DNA (D...) was then... 260 nm / D 280 nm All samples (with values ​​between 1.7 and 1.9) were sent to Sangon Biotech (Shanghai) Co., Ltd. for sequencing. PCR amplification of the V3-V4 region of the bacterial 16S rRNA gene was performed using primers 341F (5′-CCTACGGGNGGCWGCAG-3′) and 805R (5′-GACTACHVGGGTATCTAATCC-3′). Library size was detected by 2.0% agarose gel electrophoresis, and library concentration was determined using a Qubit 3.0 quantitative PCR instrument (Q33216, Thermo Fisher Scientific, China). All samples were mixed in equal volumes at a 1:1 ratio.

[0065] Table 5 shows the changes in the rumen microbiota at the phylum level. The results showed that a total of 6 phyla were identified, including Firmicutes (…). Firmicutes Bacteroidetes ( Bacteroidota ) and Proteobacteria ( ProteobacteriaFirmicutes and Actinobacteria were the dominant phyla (abundance above 10%). Compared to the CON group, the relative abundance of Firmicutes and Actinobacteria was significantly increased in the PS group. P <0.01), while the relative abundance of Proteobacteria decreased significantly ( P <0.01). Firmicutes is the dominant phylum in the rumen and plays a role in degrading crude fiber; a sustained increase in the abundance of Proteobacteria is considered a marker of intestinal flora imbalance. The abundance of Firmicutes in the Pseudomonas group was significantly higher than that in the control group, while the abundance of Proteobacteria was significantly lower than that in the control group, indicating that Pseudomonas may help promote the degradation of crude fiber and may play a role in promoting the restoration of rumen flora homeostasis.

[0066] Table 5. Changes in rumen fluid microbiota at the phylum classification level .

[0067] 3. FAPROTAX functional abundance analysis FAPROTAX (Prokaryotic Taxonomic Annotation) is a bioinformatics tool based on 16S rRNA gene sequencing data. It predicts the potential functions of microbial communities in an ecosystem by matching known microbial taxonomic information with manually compiled literature databases. In this experiment, FAPROTAX was used for functional prediction, followed by SPSS one-way ANOVA for significance analysis. Predicted functions with values ​​greater than 350 were considered the primary functions of *Pseudomonas*.

[0068] The results are as follows: FAPROTAX functional abundance analysis results are shown below. Figure 6 The FAPROTAX function predicted 52 functions, and SPASS one-way ANOVA identified 27 functions with significant differences, ultimately selecting 5 major functions. These 5 major functions are oxidative heterotrophy, fermentation, chemoheterotrophy, human digestive system, and mammalian digestive system. Compared to the CON group, the PS group showed a highly significant decrease in oxidative heterotrophic function (…). P <0.01, fermentation, chemoheterotrophy, human digestive system and mammalian digestive function significantly or extremely significantly increased ( P <0.05, P <0.01).

[0069] Although some preferred embodiments of this application have been described, those skilled in the art, upon learning the basic inventive concept, can make other changes and modifications to these embodiments. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments as well as all changes and modifications falling within the scope of this application.

[0070] Obviously, those skilled in the art can make various modifications and variations to this application without departing from the spirit and scope of its inventive concept. Therefore, if such modifications and variations fall within the scope of the claims of this application and their equivalents, this application also intends to include such modifications and variations.

Claims

1. Pseudomonas aeruginosa with accession number CCTCC M 2023814 ( Pseudomonas sp .) PSC001 is used in at least one of the following (1) to (6): (1) Degrading aflatoxin B1 or preparing preparations that degrade aflatoxin B1; (2) Promote rumen fermentation in ruminants or prepare preparations that promote rumen fermentation in ruminants; (3) Increase the production of propionic acid, butyric acid, valeric acid, isovaleric acid, and total volatile fatty acids in rumen fluid; (4) Improve the rumen microbiota; (5) Increase the relative abundance of fiber-degrading bacteria genera; (6) with Cu 2+ Combined use to enhance the degradation effect of aflatoxin B1.

2. The application according to claim 1, characterized in that, The degradation treatment conditions are: culture temperature 30 ℃~42 ℃, pH value 5.0~8.0, and treatment time 24 h~60 h.

3. The application according to claim 1, characterized in that, The concentration of *Pseudomonas* added to the rumen fermentation broth was 1 × 10⁻⁶. 9 CFU / mL ~ 8×10 9 CFU / mL.

4. A microbial agent, characterized in that, Its active ingredient is Pseudomonas aeruginosa with the preservation number CCTCC M 2023814. Pseudomonas sp PSC001, the microbial agent has the functions of degrading aflatoxin B1 and promoting rumen fermentation in ruminants.

5. The microbial agent according to claim 4, characterized in that, The bacterial agent is a liquid preparation or a lyophilized powder preparation.

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