Pediococcus pentosaceus for degrading silage fumonisins and application thereof

Abstract

The invention relates to the technical field of microorganisms, in particular to pediococcus pentosaceus for degrading silage fumonisins and application of the pediococcus pentosaceus. The pediococcus pentosaceus is preserved in the General Microbiological Culture Collection Center of the China Committee for Culture Collection of Microorganisms, and the preservation number of the pediococcus pentosaceus is CGMCC No. 28710. The pediococcus pentosaceus is high in growth rate, high in acid production efficiency, good in acid resistance and salt resistance, capable of well utilizing different types of carbon sources for fermentation, high in adaptive capacity and free of virulence genes commonly having toxic effects on livestock, the pediococcus pentosaceus serves as a pasture silage additive, the fermentation type is homotype fermentation, and the application range is wide. The pH value of the silage can be rapidly reduced, generation of ammoniacal nitrogen is reduced, the content of fumonisins is efficiently reduced, the fermentation quality of the silage is improved, and wide application prospects are achieved.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, in particular to Pediococcus pentosaceus capable of degrading silage fumonisin and application thereof. Background Art

[0002] Silage is the main roughage for dairy cows and one of the main sources of mycotoxin contamination in dairy cow diets. Fumonisins are a class of secondary metabolites produced by Fusarium fungi. They are highly toxic and can cause damage to the liver and kidneys of animals. They are also carcinogenic or teratogenic and can be transmitted to humans through meat and milk. In the process of animal breeding, the contamination of fumonisins has seriously affected the production efficiency and food safety of animal husbandry. Although there are some physical and chemical methods for the removal of fumonisins, these methods are often costly and inefficient, and may cause secondary pollution to the environment and the feed itself.

[0003] Lactic acid bacteria are the dominant bacteria in silage fermentation. They can produce organic acids to lower the pH value and inhibit undesirable microorganisms, thereby maximizing the preservation of the nutritional components of forage. At the same time, lactic acid bacteria have the advantages of being green, efficient, non-toxic, non-residue, etc., and have attracted much attention from scholars at home and abroad. It is internationally recognized as one of the most traditional and safest additives. For silage, lactic acid bacteria are efficient and safe biological control measures to inhibit mold activity and degrade fumonisin. However, there are currently few lactic acid bacteria that can inhibit mold activity and degrade fumonisin. At the same time, most of the lactic acid bacteria preparations used are derived from fermented foods, and there is a lack of lactic acid bacteria that can inhibit mold activity and degrade fumonisin for silage. Summary of the invention

[0004] In order to solve the defects existing in the prior art, the present invention provides a Pediococcus pentosaceus for degrading fumonisin in silage and its application. The Pediococcus pentosaceus has a fast growth rate, high acid production efficiency, good acid resistance and salt resistance, can be well fermented with different types of carbon sources, has strong adaptability, and does not contain common toxic genes that are toxic to livestock; the Pediococcus pentosaceus is used as a forage silage additive, and the fermentation type is homotypic fermentation, which can quickly reduce the pH value of silage, reduce the production of ammonia nitrogen, efficiently reduce the content of fumonisin, improve the fermentation quality of silage, and has broad application prospects.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a Pediococcus pentosaceus, wherein the Pediococcus pentosaceus is deposited in the General Microbiology Center of China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with a deposit number of CGMCC No. 28710 and a deposit date of October 23, 2023.

[0007] The 16s rDNA of Pediococcus pentosaceus of the present invention is shown as SEQ ID NO.1.

[0008]

[0009] In a second aspect, the present invention provides a silage additive, wherein the active ingredient of the silage additive is the above-mentioned Pediococcus pentosaceus.

[0010] Preferably, the content of Pediococcus pentosaceus in the additive accounts for 1×10 6 CFU / g.

[0011] The additives also include auxiliary materials known to those skilled in the art, and the additives may be suspensions, dispersants, or solutions.

[0012] Preferably, the silage additive is a whole-plant corn silage additive.

[0013] In a third aspect, the present invention provides a silage, wherein the silage contains the Pediococcus pentosaceus.

[0014] Preferably, the content of Pediococcus pentosaceus in the silage accounts for 1×10 6 CFU / g.

[0015] Preferably, the silage is whole-plant corn silage.

[0016] In a fourth aspect, the present invention provides an application of the Pediococcus pentosaceus, including any one of the following applications:

[0017] (a1) preparing a biological agent for degrading fumonisin;

[0018] (a2) preparing biological agents for degrading aflatoxins;

[0019] (a3) preparing a biological agent for degrading zearalenone;

[0020] (a4) preparing biological agents for degrading vomitoxin;

[0021] (a5) preparing biological agents for degrading ochratoxin;

[0022] (a6) preparing silage additives;

[0023] (a7) Preparation of silage.

[0024] In a fifth aspect, the present invention provides a method for preparing silage, comprising: mixing the Pediococcus pentosaceus with a silage raw material, and fermenting the mixture to obtain silage.

[0025] Preferably, the silage material is whole-plant corn.

[0026] Preferably, the content of Pediococcus pentosaceus in the silage accounts for 1×10 6CFU / g.

[0027] Preferably, the fermentation condition is room temperature, 25-30°C.

[0028] The beneficial effects of the present invention are:

[0029] The present invention provides a Pediococcus pentosaceus strain capable of efficiently degrading fumonisins in silage, which will be of great significance for the prevention and control of fumonisins in silage, and also provide protection for the healthy growth of livestock, food safety and human health.

[0030] When the Pediococcus pentosaceus provided by the present invention is applied to whole-plant corn silage as a silage additive, it can quickly convert sugars in silage, significantly reduce the pH value of silage, reduce the ratio of ammonia nitrogen to total nitrogen, efficiently degrade fumonisin, effectively improve the quality of silage, have good acid and salt resistance, and have great potential and broad development prospects. Further, the Pediococcus pentosaceus provided by the present invention is used as a forage silage additive, and can still well preserve the nutrients in silage after aerobic exposure for 3 days, has a low mycotoxin content, can avoid secondary fermentation of silage, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is an optical microscope image of Pediococcus pentosaceus provided by the present invention.

[0032] Figure 2 These are the results of the antibacterial activity test using the double-layer plate method in Example 1 inoculated with Pediococcus pentosaceus (left) and LPC (right).

[0033] Figure 3 Phylogenetic tree constructed for Example 2.

[0034] Figure 4 This is the growth curve of lactic acid bacteria under 37°C culture conditions in Example 3.

[0035] Figure 5 This is the acid production curve of lactic acid bacteria under 37°C culture conditions in Example 3. DETAILED DESCRIPTION

[0036] In order to enable those skilled in the art to better understand the technical solution of the invention, the present invention is further described in detail below in conjunction with specific implementation methods.

[0037] A Pediococcus pentosaceus, wherein the Pediococcus pentosaceus is deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration, the address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, the deposit number is CGMCC No. 28710, and the deposit date is October 23, 2023. The optical microscope image of the Pediococcus pentosaceus is shown in Figure 1 .

[0038] Example 1 Isolation and screening process of strains

[0039] 1. Raw materials

[0040] Soybeans: Qihuang 34 (harvested at the end of the soybean grain filling stage);

[0041] Corn: Zhongyu No. 3 (harvested at 1 / 2-2 / 3 of the milky stage);

[0042] Screening source: Corn-soybean mixed silage under 2:3 cropping pattern;

[0043] Source of Aspergillus flavus: from fresh corn;

[0044] Control strain: Commercial Lactiplantibacillus plantarum (LPC) was purchased from Sichuan Chengdu Gaofuji Biotechnology Co., Ltd.;

[0045] MRS liquid culture medium: Dissolve the raw materials (Table 1) in 1 L of distilled water, sterilize by high pressure at 121°C for 15 min, and set aside after cooling.

[0046] MRS solid medium: dissolve the raw materials (Table 1) in 1 L of distilled water, add 15 g of agar, pour into a disposable culture dish, sterilize by high pressure at 121° C. for 15 min, and set aside after cooling.

[0047] Table 1. Culture medium materials

[0048] Material Dosage Beef extract 10.00g Yeast extract 5.00g Sodium acetate anhydrous 5.00g Peptone 10.00g <![CDATA[K2HPO4]]> 2.00g Glucose 20.00g Tween-80 1.00ml <![CDATA[MgSO4.H2O]]> 0.58g Ammonium citrate 2.00g <![CDATA[MnSO4.H2O]]> 2.00g

[0049] 2. Silage samples

[0050] 2:3 corn-soybean mixed silage sample: 1 / 2-2 / 3 whole-plant corn (Zhongyu No. 3) at the milk line stage and whole-plant soybeans harvested at the late grain filling stage were placed in a 1L polyethylene silage tank in a ratio of 2:3 and stored at room temperature (30-38°C). After 60 days of silage, samples were taken from the silo for analysis.

[0051] 3. Dilution of bacterial strains

[0052] Weigh the sample into a 250 mL conical flask, add 90 mL of 0.85% sterile saline, and shake on a shaker at 120 rpm for 1 h. Draw 1 mL of the liquid and dilute it serially with sterile water (10 -1 -10 -7 ) and then spread evenly on MRS solid culture medium and culture in a 37°C constant temperature incubator for 48-72h.

[0053] 4. Initial screening of strains

[0054] According to the color, size, gloss, etc. of the colony, single colonies were picked, streaked on MRS solid medium, cultured in a 37°C constant temperature incubator for 48-72 hours, and streaked and isolated and cultured repeatedly until single colonies appeared, and enriched with MRS liquid medium (37°C, 24 hours). The lactic acid bacteria were confirmed by Gram staining and catalase test, and the lactic acid bacteria were placed in a preservation tube with glycerol at -20°C.

[0055] Using the double-layer plate culture method, a preliminary study was conducted on lactic acid bacteria that can effectively inhibit fungal growth. First, spores were scraped from the PDA solid medium that was full of spores after 5 days of culture using an inoculation loop, eluted in a triangular flask filled with 50 mL of sterile water, and the suspension was filtered into a sterile triangular flask with sterile gauze, counted with a hemocytometer, and adjusted to a concentration of 1.0×10 5 / mL. Then, inject the MRS solid culture solution into the culture dish and let it stand to solidify. Then draw two parallel lines of 2-3cm in length on the MRS plate and inoculate the strain containing Lactobacillus on it. After 24-36h of culture at 37℃, colonies can be formed. 5 PDA semi-solid culture medium with a concentration of 100 μg / mL was injected into the bottom culture medium containing lactic acid bacteria and cultured at 30°C for 48-72 hours. On this basis, antibacterial tests were performed to screen out lactic acid bacteria with antibacterial effects. Figure 2 .

[0056] Example 2 Identification of strains

[0057] 1. DNA extraction

[0058] After the strain was cultured anaerobically at 37°C in MRS liquid medium for 48 hours, the bacterial solution was transferred to a 2 mL centrifuge tube, centrifuged at 10,000 rpm for 5 minutes at 4°C, and the bacterial cells were collected. The strain DNA was extracted using the TakaRa MiniBEST Bacterial Genomic DNA Extraction Kit Ver. 3.0 (Baori Physician Technology CO., LTD, Beijing, China). The concentration of the strain DNA was measured by an ELISA instrument to determine whether it met the DNA extraction standard. The DNA extracted from the strain was stored at -20°C before use.

[0059] 2. 16S rRNA species identification

[0060] The 16S rDNA coding region was amplified by PCR. The amplification primers of 16S rDNA were 25F (5'-AAC TGAAGAGTTTGA TCC TGG CTC-3') and 1492R (5'-TAC GGC TAC CTT GTT ACG ACT-3'). The PCR reaction was 95℃ (5min), 94℃ (30s), 55℃ (1min), 72℃ (1.5min), 72℃ (10min). The sequencing was performed after 30 cycles of 94℃ (30s), 55℃ (1min), and 72℃ (1.5min). The sequence was compared with the GenBank database of the NCBI website using the nucleic acid Blast technology, and the bacterial species with the highest known taxonomic status with the sequence tested was selected. The 16S rDNA gene sequences of known strains were downloaded from the GenBank database and compared with the 16S rDNA sequences of the tested strains using Clustal. A phylogenetic tree was drawn using MEGA5.0 to determine the taxonomic status of each strain.

[0061] 3. Identification results

[0062] The strain sequencing results were compared with the homology analysis in GenBank using Nucleotide BLAST, and the strain screened had a similarity of up to 100% with Pediococcus pentosaceus. The phylogenetic tree was constructed using the Neighbor-Joining method in MAGE10.1 software ( Figure 3 ). The strain was confirmed to be Pediococcus pentosaceus by 16S rDNA sequence analysis combined with colony morphology and physiological and biochemical characteristics.

[0063] The 16s rDNA sequence of Pediococcus pentosaceus of the present invention is shown as SEQ ID NO.1.

[0064] Unless otherwise specified, the PP068 involved in the following examples of the present invention is the Pediococcus pentosaceus strain screened in Example 1.

[0065] Example 3 Determination of physiological and biochemical characteristics of strains

[0066] 1. Biological characteristics test of strains

[0067] Growth rate determination: After 48 hours of culture, the bacterial liquid of the strain was inoculated into MRS liquid medium at a 1% inoculum and cultured at 37°C for 24 hours. The OD value was measured every 2 hours at a wavelength of 600nm, and 3 groups of parallel samples were measured each time. The measurement time was 0-24 hours. After the measurement was completed, the results of the 3 groups of parallel tests were taken, the average value was calculated, and a statistical graph was drawn. The results are shown in Figure 4 .

[0068] Acid production rate determination: The strain suspension after 48 hours of culture was inoculated into MRS liquid medium with pH 6.5 at a 1% inoculation rate, and the culture temperature was 37°C and the constant temperature culture time was 24 hours. The OD value was measured at intervals of 2 hours, and 3 groups of parallel samples were measured each time and the average value was calculated. The measurement time was 0-24 hours. After the root measurement was completed, the data of the three tests were compared, and the average value was calculated. The acid production rate curve was drawn. The results are shown in Figure 5 .

[0069] from Figure 4 It can be seen that compared with the commercial strain of Lactobacillus plantarum, at 37°C, the Pediococcus pentosaceus of the present invention begins to grow rapidly at the 6th hour and enters the logarithmic phase, reaches a maximum value at the 16th hour, and then is in a stable phase; the strain LPC begins to grow rapidly at the 2nd hour, the growth rate reaches a maximum value at the 12th hour, and then the growth rate decreases slightly, and grows slowly again after the 16th hour, with small fluctuations.

[0070] from Figure 5 It can be seen that compared with the commercial strain of Lactobacillus plantarum, the pH value of the bacterial solution of the present invention rapidly decreases after fermentation for 2-8 hours at 37°C, and the acid production rate is fast; after culturing for 8 hours, the pH value slowly decreases, indicating that the acid production rate of the strain gradually decreases; after culturing for 18 hours, the pH value is reduced to 3.9, and the pH value remains basically unchanged thereafter. The acid production rate curve is basically consistent with that of the control group LPC, and the pH value after 24 hours is basically the same.

[0071] 2. Salt and acid resistance of strains

[0072] The correspondence between OD value and growth condition is shown in Table 2.

[0073] Table 2. OD value and growth status

[0074]

[0075] Acid resistance: Use 0.1 mol / L HCl or NaOH to adjust the pH value of MRS liquid medium (3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 8.0). In the same way as above, inoculate 1% of the inoculum into MRS liquid medium, plug the test tube, shake evenly, place the bacterial solution in a 37°C constant temperature incubator for 48-72 hours, observe the growth of the strain and measure the OD 600 The absorbance is shown in Table 3.

[0076] Salt tolerance: The activated lactic acid bacteria were inoculated into MRS liquid medium containing 3% and 6.5% NaCl at a 1% inoculation rate, and the bacterial solution was cultured in a 37°C incubator for 72 h. The growth of the strain was observed and the OD was measured. 600 The absorbance is shown in Table 3.

[0077] Temperature resistance: The activated lactic acid bacteria strains were inoculated into MRS liquid culture medium at a rate of 1%, and the culture solution was placed in an incubator at 30°C, 37°C, 45°C, and 50°C for 48-72 hours. The growth of the strains was observed and the OD was measured. 600 The absorbance is shown in Table 3.

[0078] Table 3. Physiological and biochemical characteristics of Pediococcus pentosaceus

[0079]

[0080] In Table 3, +: normal growth; ++: good growth; Homo: homofermentative strain; Hetero: heterofermentative strain; LR: Pediococcus pentosaceus standard strain; LPC: Lactobacillus plantarum commercial strain.

[0081] As shown in Table 3, the Pediococcus pentosaceus provided by the present invention is a Gram-positive, homofermentative coccus, grows well under the culture conditions of 50°C, 3.0% and 6.5% NaCl concentration and pH 3.0-8.0, and has good temperature resistance, salt resistance and acid resistance.

[0082] 3. Sugar fermentation spectrum test of strains

[0083] Sugar fermentation: HBI microbial biochemical identification strips (Qingdao Hi-Tech Industrial Park Haibo Technology Co., Ltd.) were used to conduct identification tests on the fermentation of different sugars and alcohols. The fermentation of different sugars and alcohols by Pediococcus pentosaceus was analyzed through sugar fermentation tests. The results are shown in Table 4.

[0084] Table 4. Sugar fermentation characteristics of Pediococcus pentosaceus

[0085]

[0086] In Table 4, +: growth; -: no growth; LR: standard strain of Lactobacillus rhamnosus; LPC: commercial strain of Lactobacillus plantarum.

[0087] As shown in Table 4, the Pediococcus pentosaceus provided by the present invention can utilize a wide range of carbon sources and can better utilize different types of carbon sources for fermentation. The Pediococcus pentosaceus with antibacterial properties can utilize esculin, cellobiose, maltose, salicin, sucrose and inulin, all of which are positive, but cannot utilize hippuric acid, mannitol, sorbitol, raffinose and lactose.

[0088] 4. Detection of virulence genes that inhibit Aspergillus flavus strains

[0089] The detection of virulence genes of strains is mainly through serotype typing and detection of common virulence genes. There are 11 common virulence genes, including nheA, nheB, entFM, bceT, hblA, hblB, hblC, hblD, ces, cer, and cytK.

[0090] Table 5. Primers for virulence gene detection

[0091]

[0092]

[0093] As shown in Table 5, the nheA, nheB, entFM, bceT, hblA, hblB, hblC, hblD, ces, cer, and cytK virulence genes were not detected in the amplification of the virulence genes of lactic acid bacteria that inhibit Aspergillus flavus.

[0094] Example 4 Detection of silage

[0095] 1. Preparation of silage: Cut the whole plant of Zhongyu No. 3 corn (harvested at the milk line stage) into 2-3 cm pieces with a guillotine and cut into 1×10 6 The CFU / g content requires that the lactic acid bacteria additive and the whole corn be mixed evenly and then packed into silage bags (specifications are 30cm*28cm), vacuum sealed with a vacuum machine, and opened after fermentation at room temperature (25-30℃) for 0d and 60d. This silage process is carried out in a constant temperature environment.

[0096] 2. Fermentation quality and chemical composition analysis

[0097] Before sampling, the silage feed must be fully mixed. 200g of fresh samples, samples of silage for 60 days or aerobic exposure samples are taken, dried at 65℃ for 72h (DL-101-2BS electric blast drying oven, Tianjin Zhonghuan Experimental Electric Furnace Co., Ltd.), and then weighed to measure the dry matter (DM) content; the dried samples are crushed and sieved (60 mesh) for determination of chemical composition. Take 10g of sample, dilute 10 times with 90mL of sterile water, and extract overnight in a refrigerator at 4℃. Filter with sterile cloth and take the supernatant for determination of fermentation quality. The above operations are repeated three times. Fermentation quality includes pH and organic acid content, and each index is determined as follows: the pH of the extract is determined using a Leiji PHS-3C precision pH meter; the organic acid analysis method (SHIMADZE-10A high performance liquid chromatograph, chromatographic column: ShodexRspak KC-811S-DVB gel Column 300×8mm, detector: SPD-M10AVP, mobile phase: 3mmol·L-1 perchloric acid, flow rate: 1mL·min-1, injection volume: 5μL, column temperature: 50℃, detection wavelength: 210nm) is used to determine the content of organic acids (lactic acid (LA), acetic acid (AA), propionic acid (PA) and butyric acid (BA)). The prepared dry powder was taken and the nutrient content was determined. The crude protein (CP) content was determined by the Kjeldahl method; the neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents were determined by the Van der Waals fiber washing method; the soluble carbohydrate (WSC) content was determined by the anthrone colorimetric method.

[0098] Table 6. Nutritional composition of fresh whole corn

[0099]

[0100] The chemical composition of fresh whole-plant corn before ensilage is shown in Table 6. The DM of fresh corn samples was 33.97%, and the concentrations of CP, NDF, ADF and WSC were 8.46%, 29.74%, 13.74% and 9.42%, respectively. The concentrations of aflatoxin B1, aflatoxin B2, zearalenone, vomitoxin, ochratoxin, fumonisin B1, fumonisin B2 and fumonisin B3 were 21.52, 4.58, 1055.19, 4.11, 3.71, 1852.58, 517.04 and 127.71 μg / kg, respectively.

[0101] Table 7. Fermentation quality of whole-plant corn silage during aerobic exposure

[0102]

[0103] Note: Different letters indicate significant differences (P<0.05). SEM, standard error; * indicates significant difference (P<0.05); ** indicates extremely significant difference (P<0.01); NS, not significant; T, aerobic exposure time of silage; E, additive; T×E, time×additive. CK, control group; AF, Aspergillus flavus group; AFLPC, Aspergillus flavus + commercial Lactobacillus plantarum; AFPP068, Aspergillus flavus + Pediococcus pentosaceus 068.

[0104] As can be seen from Table 7, in the early stage of aerobic exposure of whole-plant corn silage (0-3d), the pH and AN / TN values ​​of the AFPP608 and AFLPC groups were significantly lower than those of the CK and AF groups (P<0.05), and the acetic acid content was significantly higher than that of the CK and AF groups, and the difference between the two groups was not significant (P>0.05). In addition, compared with other treatment groups, the lactic acid content was significantly increased by inoculation of PP068 (P<0.05), and no butyric acid was detected during the aerobic exposure period, indicating good silage quality.

[0105] 3. Mycotoxin content analysis

[0106] Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS) was used to determine the contents of aflatoxins (AFBS), zearalenone (ZEA), deoxynivalenol (DON), ochratoxins (OT) and fumonisins (FBS). The mycotoxin concentrations of fresh corn samples are shown in Table 6; the mycotoxin concentrations during aerobic exposure are shown in Table 8.

[0107] Table 8. Mycotoxin levels in whole corn silage

[0108]

[0109]

[0110] Note: Different letters indicate significant differences (P< 0.05). SEM, standard error; * indicates significant difference (P<0.05); ** indicates extremely significant difference (P<0.01); NS, not significant; T, silage aerobic exposure time; E, additive; T×E, time×additive. CK, control group; AF, Aspergillus flavus group; AFLPC, Aspergillus flavus + commercial Lactobacillus plantarum; AFPP068, Aspergillus flavus + Pediococcus pentosaceus 068.

[0111] Table 8 shows the concentrations of aflatoxin B1, aflatoxin B2, zearalenone, vomitoxin, ochratoxin, fumonisin B1, fumonisin B2 and fumonisin B3 in whole corn silage after adding Pediococcus pentosaceus of the present invention at 0 days (60 days of silage) and 3 days of oxygen exposure. Compared with fresh corn, the concentrations of aflatoxin B1, aflatoxin B2, zearalenone, vomitoxin, ochratoxin, fumonisin B1, fumonisin B2 and fumonisin B3 in the added treatment group at 0 days (60 days of silage) and 3 days of oxygen exposure were all reduced. At 0 day of aerobic exposure (60 days of ensiling), the contents of aflatoxin B1 (4.35 μg / kg), aflatoxin B2 (1.44 μg / kg), zearalenone (383.78 μg / kg), vomitoxin (1.56 μg / kg), ochratoxin (1.57 μg / kg), fumonisin B1 (463.77 μg / kg), fumonisin B2 (162.2 μg / kg) and fumonisin B3 (60.41 μg / kg) in the AFPP068 group were significantly lower than those in the AF group (34.58%, 33.02%, 21.70%, 9.82%, 19.49%, 56.12%, 52.28% and 30.16%), with significant differences (P < 0.01). After 3 days of aerobic exposure, the content of zearalenone in the AFPP068 group was still significantly lower than that in the other treatment groups (P<0.01). In addition, except for the AFPP068 group, the zearalenone content in the other treatment groups showed a rapid increasing trend and exceeded the maximum limit standard of Chinese dairy cow feed (500 μg / kg).

[0112] The above are only preferred embodiments of the present invention. It should be noted that the above preferred embodiments should not be regarded as limiting the present invention, and the protection scope of the present invention should be based on the scope defined by the claims. For ordinary technicians in this technical field, several improvements and modifications can be made without departing from the spirit and scope of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. A Pediococcus pentosaceus, characterized in that The Pediococcus pentosaceus is deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with the deposit number being CGMCC No.28710.

2. The Pediococcus pentosaceus according to claim 1, characterized in that The 16s rDNA of Pediococcus pentosaceus is shown as SEQ ID NO.

1.

3. A silage additive, characterized in that: The active ingredient of the silage additive is the Pediococcus pentosaceus according to claim 1 or 2.

4. A silage, characterized in that: The silage contains the Pediococcus pentosaceus according to claim 1 or 2.

5. The silage according to claim 4, characterized in that: The content of Pediococcus pentosaceus in the silage feed accounts for 1×10 6 CFU / g.

6. The silage according to claim 4, characterized in that: The silage is whole-plant corn silage.

7. A use of Pediococcus pentosaceus according to claim 1, characterized in that: Includes any of the following applications: (a1) preparing a biological agent for degrading fumonisin; (a2) preparing biological agents for degrading aflatoxins; (a3) preparing a biological agent for degrading zearalenone; (a4) preparing biological agents for degrading vomitoxin; (a5) preparing biological agents for degrading ochratoxin; (a6) preparing silage additives; (a7) Preparation of silage.

8. A method for preparing silage, characterized in that: include: The Pediococcus pentosaceus according to claim 1 or 2 is mixed with silage raw materials and fermented to obtain silage.

9. The preparation method according to claim 8, characterized in that: The ratio of the amount of Pediococcus pentosaceus added to the silage raw material is 1×10 6 CFU / g.

10. The preparation method according to claim 8, characterized in that: The fermentation temperature is 25-30°C.

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