A strain of Pediococcus acidilactici, a composition containing the same and application thereof

Abstract

The present invention relates to the field of microbial technology, and in particular to a strain of Pediococcus acidilactici, a composition containing the same, and uses thereof. The Pediococcus acidilactici is Pediococcus acidilactici H47, with a deposit number of CCTCC NO: M20232100, and is deposited in the China Center for Type Culture Collection. The composition comprises at least one of Pediococcus acidilactici and a culture of the strain. The advantages of the present invention are as follows: (1) efficient degradation of mycotoxins. (2) strong growth ability, acid and bile resistance. (3) safety, and good probiotic properties.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a strain of Pediococcus acidilactici H47, a composition containing the same and uses thereof. Background Art

[0002] Feed and its raw materials are prone to mildew and mold during processing, storage and transportation. Mold not only reduces the nutritional value of feed, affects palatability, and leads to a decrease in livestock and poultry feed intake, but also causes animal growth stagnation, reduced immunity, decreased production performance, and even poisoning and death. According to a recent global survey, 90% of feeds and raw materials contain at least one mycotoxin, and 64% of feeds are contaminated with multiple mycotoxins. The positive detection rates of aflatoxin B1 (AFB1), zearalenone (ZEA), and deoxynivalenol (DON) in corn samples collected from multiple regions in China were 94.9%, 93.4%, and 98.4%, respectively. Among them, 92.4% of the samples were detected with these three toxins at the same time, indicating that co-contamination with multiple mycotoxins is very common.

[0003] Common mycotoxins in daily life include AFB1, ZEA, DON, T-2 toxin (T-2), ochratoxin (OTA), fumonisin B1 (FB1), cyclopiazonic acid (CPA), etc. Different mycotoxins have different sources, physiological reactions and clinical symptoms: AFB1 originates from Aspergillus flavus, which can damage animal livers and suppress immune function. Long-term exposure can cause cancer and death in animals and humans. ZEA is an estrogenic toxin produced by Fusarium. The symptoms of poisoning are reduced feed conversion rate, changes in organ weight, decreased animal fertility and abnormal behavior. DON and T-2 come from the genus Fusarium, which are type B and type A Trichoderma, respectively. They can directly damage the mucosa of the animal digestive tract, generally manifested as loss of appetite, bleeding, vomiting, diarrhea, etc. OTA originates from Aspergillus and Penicillium, and is an immunosuppressant, teratogen and nephrotoxic compound. Poisoning can cause lesions such as renal cortical degeneration and medullary hemorrhage, and its target organ is the kidney.

[0004] At present, diseases caused by bacteria and fungi are generally treated with antibiotics, but the abuse of antibiotics has caused most bacteria and fungi to develop varying degrees of resistance to them. The phenomenon of drug residues in food has become a serious public health and food safety problem. At the same time, the state issued an announcement that "from July 1, 2020, antibiotics will be completely banned in livestock and poultry feed." After the ban on antibiotics in feed, it is a new round of challenges facing the livestock and poultry farming industry after the African swine fever epidemic. Therefore, finding beneficial alternatives to antibiotics has become an urgent problem to be solved. Lactic acid bacteria can be used as a potential alternative to antibiotics because of its advantages such as high detoxification efficiency, strong specificity and no pollution. At the same time, lactic acid bacteria can also improve the balance of intestinal microecology, promote animal growth and development, enhance the body's immunity, fight bacteria and prevent diseases, and have nutritional health functions. Summary of the invention

[0005] The purpose of the present invention is to provide a Pediococcus acidilactici that can efficiently degrade mycotoxins, has probiotic properties, strong growth ability, acid and bile resistance, and is non-hemolytic, as well as a composition containing the same and an application thereof.

[0006] Storage Instructions:

[0007] Taxonomic nomenclature: Pediococcus acidilactici H47;

[0008] Deposit time: November 1, 2023;

[0009] Depository: China Center for Type Culture Collection; Address: No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province

[0010] Deposit number: CCTCC NO: M20232100.

[0011] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0012] In a first aspect, the present invention provides a Pediococcus acidilactici.

[0013] The invention discloses Pediococcus acidilactici, which is Pediococcus acidilactici H47, with a preservation number of CCTCC NO: M20232100, and is preserved in China Center for Type Culture Collection.

[0014] In a second aspect, the present invention provides a microbial preparation.

[0015] A microbial preparation comprising Pediococcus acidilactici or a culture of the strain.

[0016] The Pediococcus acidilactici is Pediococcus acidilactici H47, with a preservation number of CCTCC NO: M20232100, and is preserved in China Center for Type Culture Collection.

[0017] In a third aspect, the present invention provides a composition.

[0018] A composition comprising at least one of Pediococcus acidilactici and a culture of said strain;

[0019] The culture of the strain includes but is not limited to the fermentation broth, fermentation precipitation or freeze-dried powder of Pediococcus acidilactici.

[0020] The Pediococcus acidilactici is Pediococcus acidilactici H47, with a preservation number of CCTCC NO: M20232100, and is preserved in China Center for Type Culture Collection.

[0021] Furthermore, the composition is a microecological preparation or a pharmaceutical preparation.

[0022] Furthermore, the composition is a probiotic.

[0023] Furthermore, the composition also contains substances that help maintain the vitality of Pediococcus acidilactici; the composition also contains other probiotics and / or prebiotics.

[0024] In a fourth aspect, the present invention provides use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect.

[0025] Use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect in preparing a composition for degrading mycotoxins.

[0026] In a fifth aspect, the present invention provides another use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect.

[0027] Use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect in preparing a composition for a probiotic.

[0028] In a sixth aspect, the present invention provides a third use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect.

[0029] Use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect in preparing a composition for feed fermentation.

[0030] In the fourth, fifth and sixth uses, the Pediococcus acidilactici is Pediococcus acidilacticiH47, with a preservation number of CCTCC NO: M20232100, which is preserved in China Center for Type Culture Collection.

[0031] In a seventh aspect, the present invention provides an animal feed.

[0032] An animal feed, comprising the Pediococcus acidilactici of the first aspect;

[0033] and / or a fermentation product of Pediococcus acidilactici according to the first aspect;

[0034] and / or the microbial preparation of the second aspect;

[0035] and / or the composition of the third aspect.

[0036] The composition of Pediococcus acidilactici provided by the present invention may contain a substance that helps to maintain the vitality of Pediococcus acidilactici. The substance that helps to maintain the vitality of Pediococcus acidilactici (such as a protective agent) may be any one of cysteine, glutathione, butylated hydroxyanisole, dibutylmethyltoluene, tocopherol, bamboo leaf antioxidants, D-isoascorbic acid and its sodium salt, sodium ascorbate, calcium ascorbate, phospholipids, vitamin C (ascorbic acid), and vitamin E, or a combination thereof.

[0037] The Pediococcus acidilactici composition provided by the present invention may contain other probiotics, and the probiotics may be selected from any one of lactic acid bacteria, bifidobacteria, and Lactobacillus acidophilus, or a combination thereof.

[0038] The composition of Pediococcus acidilactici provided by the present invention may contain prebiotics, and the prebiotics may be selected from any one of fructooligosaccharides (FOS), galacto-oligosaccharides (GOS), xylooligosaccharides (XOS), lactofructooligosaccharides (LACT), soybean oligosaccharides (SOS), inulin, and oligosaccharides, or a combination thereof.

[0039] The lactic acid Pediococcus provided by the present invention is Pediococcus acidilactici H47, with a deposit number of CCTCCNO: M20232100, and was deposited in the China Center for Type Culture Collection on November 1, 2023. The strain was isolated from the rumen of Tibetan sheep in Qinghai. Lactic acid Pediococcus (Pediococcus acidilactici H47) can efficiently degrade mycotoxins and has probiotic properties; it can be used as a feed additive and as a strain fermented feed, and at the same time can improve the utilization rate of feed.

[0040] The Pediococcus acidilactici H47 provided by the present invention can efficiently degrade mycotoxins, and the degradation rates of AFB1 (Aaflatoxin B1, aflatoxin B1), DON (Deoxynivalenol, vomitoxin), ZEA (Zearalenone, zearalenone), and GLUS (glucosinolate) by Pediococcus acidilactici H47 are 64.16%, 66.06%, 45.40%, and 75.37%, respectively. The average degradation rate of AFB1, DON, ZEA, and GLUS by Pediococcus acidilactici H47 is 64.32%.

[0041] The Pediococcus acidilactici H47 provided by the invention enters a logarithmic growth phase 4 hours after inoculation, has a short fermentation time, strong environmental adaptability and vigorous vitality, can grow normally in an acidic environment of pH 3.5, is completely tolerant to a bile salt environment of 0.1%, has high tolerance to acid and bile salts, and is completely tolerant to a NaCl concentration of 2.0%.

[0042] The Pediococcus acidilactici H47 provided by the present invention has no hemolytic symptoms, is moderately sensitive to erythromycin, is insensitive to streptomycin and gentamicin, is moderately sensitive to penicillin, is insensitive to tetracycline, and is insensitive to polymyxin B. It has a strong inhibitory effect on Salmonella, Escherichia coli, and has a very strong inhibitory effect on Staphylococcus aureus. This shows that the strain is not only safe, but also has good probiotic properties.

[0043] The present invention separates and screens lactic acid bacteria that can efficiently degrade mycotoxins and have probiotic properties from the rumen of Qinghai Tibetan sheep. The lactic acid bacteria ferment feed, which can not only degrade multiple fungal toxins in feed raw materials and improve the safety of feed, but also increase the utilization of some unconventional feed raw materials. The invention has a very good prospect in the application of fermented feed with unconventional feed as raw material.

[0044] Compared with the prior art, the present invention provides a strain of Pediococcus acidilactici, a composition containing the same and the advantages of its use:

[0045] (1) Highly efficient degradation of mycotoxins.

[0046] (2) Strong growth ability and resistance to acid and bile salts.

[0047] (3) It is safe and has good probiotic properties. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1The present invention provides a Pediococcus acidilactici H47 strain colony morphology (left figure) and strain Gram staining (right figure).

[0049] Figure 2 The present invention provides an agarose gel electrophoresis diagram of the genomic DNA of Pediococcus acidilactici H47.

[0050] Figure 3 The present invention provides an electrophoresis diagram of the product of PCR amplification of the genome DNA of Pediococcus acidilactici H47.

[0051] Figure 4 The present invention provides a phylogenetic tree of Pediococcus acidilactici H47.

[0052] Figure 5 The present invention provides a growth curve of Pediococcus acidilactici H47.

[0053] Figure 6 It is the acid production curve of Pediococcus acidilactici H47 provided by the present invention.

[0054] Figure 7 The present invention provides the growth of Pediococcus acidilactici H47 under different pH conditions.

[0055] Figure 8 The invention provides the growth conditions of Pediococcus acidilactici H47 under different bile salt concentration conditions.

[0056] Fig. 9 The present invention provides the growth conditions of Pediococcus acidilactici H47 under different concentrations of NaCl.

[0057] Fig.10 It is the result of the hemolysis test of Pediococcus acidilactici H47 provided by the present invention.

[0058] Fig.11 The present invention provides a Pediococcus acidilactici H47 drug sensitivity test result to 8 antibiotics.

[0059] Fig.12 The detoxification curves of Pediococcus acidilactici H47 provided by the present invention to AFB1, DON, ZEA and GLUS.

[0060] Figure 2 and Figure 3 In the figure, M represents DL 2000 Marker; Band 6 represents Pediococcus acidilactici H47.

[0061] Fig.11 In the table, 1 represents polymyxin B; 2 represents ciprofloxacin; 3 represents tetracycline; 4 represents gentamicin; 5 represents erythromycin; 6 represents ofloxacin; 7 represents penicillin; and 8 represents streptomycin. DETAILED DESCRIPTION

[0062] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention is further described in detail in the following examples. The following examples are only used to illustrate the invention but are not used to limit the scope of the present invention.

[0063] In a first aspect, the present invention provides a Pediococcus acidilactici.

[0064] The invention discloses Pediococcus acidilactici, which is Pediococcus acidilactici H47, with a preservation number of CCTCC NO: M20232100, and is preserved in China Center for Type Culture Collection.

[0065] In a second aspect, the present invention provides a microbial preparation.

[0066] A microbial preparation comprising Pediococcus acidilactici or a culture of the strain.

[0067] The Pediococcus acidilactici is Pediococcus acidilactici H47, with a preservation number of CCTCC NO: M20232100, and is preserved in China Center for Type Culture Collection.

[0068] In a third aspect, the present invention provides a composition.

[0069] A composition comprising at least one of Pediococcus acidilactici and a culture of said strain;

[0070] The culture of the strain includes but is not limited to the fermentation broth, fermentation precipitation or freeze-dried powder of Pediococcus acidilactici.

[0071] The Pediococcus acidilactici is Pediococcus acidilactici H47, with a preservation number of CCTCC NO: M20232100, and is preserved in China Center for Type Culture Collection.

[0072] Furthermore, the composition is a microecological preparation or a pharmaceutical preparation.

[0073] Furthermore, the composition is a probiotic.

[0074] Furthermore, the composition also contains substances that help maintain the vitality of Pediococcus acidilactici; the composition also contains other probiotics and / or prebiotics.

[0075] In a fourth aspect, the present invention provides use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect.

[0076] Use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect in preparing a composition for degrading mycotoxins.

[0077] In a fifth aspect, the present invention provides another use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect.

[0078] Use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect in preparing a composition for a probiotic.

[0079] In a sixth aspect, the present invention provides a third use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect.

[0080] Use of the Pediococcus acidilactici of the first aspect, the microbial preparation of the second aspect, or the composition of the third aspect in preparing a composition for feed fermentation.

[0081] In the fourth, fifth and sixth uses, the Pediococcus acidilactici is Pediococcus acidilacticiH47, with a preservation number of CCTCC NO: M20232100, and is preserved in the China Center for Type Culture Collection.

[0082] In a seventh aspect, the present invention provides an animal feed.

[0083] An animal feed, comprising the Pediococcus acidilactici of the first aspect;

[0084] and / or a fermentation product of Pediococcus acidilactici according to the first aspect;

[0085] and / or the microbial preparation of the second aspect;

[0086] and / or the composition of the third aspect.

[0087] Example 1

[0088] Isolation and Identification of Pediococcus acidilactici H47

[0089] 1. Morphological identification

[0090] The lactic acid bacteria strains isolated from the rumen of Qinghai Tibetan sheep were inoculated in MRS solid and liquid culture media, respectively, and cultured at 37°C for 24 hours to observe the size, shape, surface characteristics, edge shape, color, transparency, etc. of the single colony. An inoculation loop was used to pick up an appropriate amount of culture in the center of the slide, and after careful coating, a coverslip was placed on it to observe the morphology and size of the strains under an optical microscope.

[0091] like Figure 1 As shown in Table 1, the strains obtained by screening are suspected to be lactic acid bacteria strains. The Gram staining test was positive and confirmed to be Gram-positive bacteria. The catalase test reactions were all negative. Gram-positive bacteria are purple in color under a microscope, and the strains are spherical and rod-shaped. The strains are white; the strains are round and slightly raised, and the colony surface is smooth and moist with neat edges.

[0092] Table 1 Morphological characteristics of lactic acid bacteria

[0093]

[0094] “+”: positive; “-”: negative.

[0095] 2. Physiological and biochemical identification

[0096] Some physiological and biochemical experiments were carried out using a bacterial physiological and biochemical identification kit, including esculin, mannitol, sucrose, maltose, cellobiose, salicin, sorbitol, and raffinose; the catalase test, motility test, hydrogen sulfide test, glucose gas production test, gelatin liquefaction test, and indole test were used to preliminarily identify the strains with reference to the Bergey's Manual of Systematic Bacteriology GB4789.35-201645.

[0097] As shown in Table 2, the test showed that the strain did not produce gas, was negative in the indole test, catalase test, and hydrogen sulfide test, and did not liquefy gelatin. The final identification result was determined by combining the results of morphological analysis and 16S rRNA gene fragment sequence analysis.

[0098] Table 2 Physiological and biochemical characteristics of lactic acid bacteria

[0099]

[0100] “+”: positive; “-”: negative.

[0101] 3. Molecular Biological Identification

[0102] The OMEGA bacterial genomic DNA extraction kit was used to extract the lactic acid bacteria genomic DNA. The 16S rRNA gene amplification primers were synthesized by Shanghai Shenggong Biotechnology Co., Ltd., and the sequences are as follows: 16S F: 5'-AGA GTT TGA TCC TGG CTCAG-3'; 16S R: 5'-GGT TAC CTT GTT ACG ACT T-3'. The genomic DNA prepared above was used as a template for PCR amplification, and PCR amplification was performed using a 50μL reaction system. The cycle parameters were as follows: 95℃ pre-denaturation for 1min, 95℃ denaturation for 30s, 55℃ annealing for 30s, 72℃ extension for 2min, 72℃ end extension for 5min, 30 cycles, and the amplified products were sequenced. The sequencing results were spliced ​​and compared in BLAST. The typical strain sequences with a similarity of more than 97.00% were selected, and the phylogenetic tree was constructed using MEGA software.

[0103] like Figure 2 and Figure 3 As shown in the figure, the amplified fragment was detected by electrophoresis and gel imaging analysis, and a single fluorescent band appeared at 1500bp. The amplified product had good integrity, no degradation, no tailing, and no non-specific amplification, which met the sequencing requirements. The sequencing results were compared by Blast and the phylogenetic tree was constructed ( Figure 4 ) found that H47 is lactic acid Pediococcus. From Table 3 and Figure 4 Identify H47 Pediococcus acidilactici.

[0104] Table 3 Results of homology comparison of lactic acid bacteria 16S rRNA sequences

[0105]

[0106] Example 2

[0107] Probiotic properties of Pediococcus acidilactici H47

[0108] 1. Determination of growth ability and acid production capacity

[0109] The bacterial suspension of Pediococcus acidilactici H47 was inoculated into MRS liquid medium at a 3% inoculation rate and cultured at 37°C. The OD values ​​were measured at 0h, 2h, 4h, 6h, 8h, 12h, 16h, 24h, 36h, and 48h, with 3 replicates in each group. The blank medium was used as the control. 600 The growth curve and acid production curve were drawn with the culture time of the bacterial solution as the horizontal axis and the absorbance value and pH value as the vertical axis.

[0110] like Figure 5 As shown in the growth curve, the H47 strain is in a sluggish phase between 0 and 4 hours, and enters a logarithmic growth phase 4 hours after inoculation. The fermentation time is short, the environmental adaptability is strong, and the vitality is vigorous; the strain is in a stable phase between 24 and 48 hours, and the OD at 48 hours is 600 The value is 1.62, which has strong fermentation activity.

[0111] like Figure 6 The acid production curve shows that the pH of the H47 strain enters a rapid decline phase from a stable state and then gradually tends to a stable phase. When the strain enters the logarithmic growth phase, the pH of the medium begins to decline. After the strain enters the stable growth phase, the pH begins to stabilize again. The H47 strain enters the logarithmic phase at 4h, and the pH value at 48h is below 4.5, indicating good acid production performance.

[0112] 2. Acid and bile salt tolerance

[0113] The bacterial suspension of Pediococcus acidilactici H47 was inoculated into MRS liquid medium with pH values ​​of 2.5, 3.0, 3.5, 4.0, and 6.0 at a 1% inoculation rate, and cultured at 37°C for 24 h. The OD of the bacterial suspension was measured. 600 The inoculum was inoculated into MRS liquid medium with 0.0%, 0.1%, 0.2%, and 0.3% ox bile salt concentrations at a 1% inoculum volume, and the OD of the bacterial solution was measured for 24 hours. 600 The blank medium with the same acidity and bile salt concentration was used as the control.

[0114] like Figure 7 As shown, Pediococcus acidilactici H47 can basically grow normally in the acidic environment of pH 3.5 and pH 4.0, but is basically intolerant to the acidic environment of pH 3.0 and pH 2.5 and its growth stagnates.

[0115] like Figure 8 As shown, Pediococcus acidilactici H47 is completely tolerant to 0.1% bile salt environment; under 0.2% bile salt conditions, the growth of Pediococcus acidilactici H47 is inhibited.

[0116] 3. Osmotic pressure resistance

[0117] 2.0%, 4.0%, 6.0%, and 8.0% NaCl were added to the MRS liquid culture medium, and the bacterial suspension of Pediococcus acidilactici H47 was inoculated into the MRS liquid culture medium with different NaCl concentrations at a 1% inoculation rate. After constant temperature culture at 37°C for 24 h, the OD of each group of bacterial suspension was measured. 600 Each group was repeated 3 times, and blank medium with the same NaCl concentration was used as control.

[0118] like Fig. 9As shown, Pediococcus acidilactici H47 was completely tolerant to 2.0% NaCl concentration, its OD value slowly decreased at 4.0% NaCl concentration, and was inhibited in an environment with NaCl concentration higher than 6.0%.

[0119] 4. Safety inspection

[0120] (1) Lactic acid bacteria hemolysis test

[0121] All screened strains were streaked on Columbia blood plates and incubated for 24 to 48 hours. Observe whether there are hemolytic zones around the colonies: α-hemolysis, β-hemolysis, and γ-hemolysis, and take photos and record.

[0122] like Fig.10 As shown, Pediococcus acidilactici H47 showed no hemolytic symptoms (γ hemolysis) after 48 h of incubation.

[0123] (2) Lactic acid bacteria antibiotic sensitivity test

[0124] The drug sensitivity test of 8 antibiotics was carried out on the H47 strain of Pediococcus acidilactici using the drug sensitivity paper method, including Polymycin B (PB); Ciorofloxacin (CIP); Tetracycline (TE); Gentamicin (GEN); Erythromycin (E); Ofloxacin (OFX); Penicillin (P); Streptomycin (S).

[0125] like Fig.11 As shown, Pediococcus acidilactici H47 was moderately sensitive to erythromycin, insensitive to streptomycin and gentamicin, moderately sensitive to penicillin, insensitive to polymyxin B, and insensitive to ciprofloxacin.

[0126] (3) Determination of antibacterial activity of lactic acid bacteria

[0127] The Oxford cup method was used to determine the diameter of the inhibition zone of the fermentation supernatant of the strain against pathogenic bacteria such as Escherichia coli, Salmonella, and Staphylococcus aureus.

[0128] As shown in Table 4, Pediococcus acidilactici H47 has a strong inhibitory effect on Salmonella, Escherichia coli and , and has a very strong inhibitory effect on Staphylococcus aureus.

[0129] Table 4 Antibacterial results of lactic acid bacteria

[0130]

[0131] "+" indicates that the diameter of the antibacterial zone is 9.0-12.99 mm; "++" indicates that the diameter of the antibacterial zone is 13.0-16.99 mm; "+++" indicates that the diameter of the antibacterial zone is 17.0-19.99 mm; "++++" indicates that the diameter of the antibacterial zone is ≥20 mm

[0132] Example 3

[0133] Degradation of mycotoxins by Pediococcus acidilactici H47

[0134] 1. Mycotoxin degradation rate test

[0135] The degradation effect of Pediococcus acidilactici H47 on AFB1, DON, ZEA and GLUS was tested, and the degradation rates of the four mycotoxins were determined after 72 h of fermentation. The results are shown in Table 5.

[0136] As shown in Table 5, the degradation rate of AFB1 by Pediococcus acidilactici H47 was 70.95%, the degradation rate of DON was 13.50%, the degradation rate of ZEA was 34.11%, and the degradation rate of GLUS was 48.44%.

[0137] Table 5 Degradation rate of four toxins by lactic acid bacteria

[0138]

[0139] 2. Lactic acid bacteria detoxification curve

[0140] Pediococcus acidilactici H47 was inoculated at 1% in MRS liquid culture medium and cultured at 37°C for 24 hours, then cultured with the four toxins respectively. The toxin contents were measured at 0h, 2h, 4h, 8h, 12h, 1d, 3d, 7d, and 14d. A detoxification curve was drawn with the culture time as the horizontal axis and the degradation rate as the vertical axis to analyze the changes in toxin degradation rate in different time periods.

[0141] like Fig.12 As shown in the figure, the degradation rates of the four toxins by lactic acid Pediococcus H47 increased with time. The degradation rate of AFB1 increased slowly before 12 hours, increased rapidly after 12 hours, and gradually tended to a stable stage after 7 days. The degradation rate of DON increased slowly before 12 hours, and gradually tended to a stable stage after 12 hours. The degradation rate of ZEA increased rapidly before 8 hours, tended to be flat between 8 and 12 hours, and increased rapidly between 12 and 24 hours. At 14 days of fermentation, the degradation rates of AFB1, DON, ZEA and GLUS by lactic acid Pediococcus H47 were 64.16%, 66.06%, 45.40% and 75.37%, respectively.

[0142] In summary, with the increase of fermentation time, the degradation rate of AFB1, DON, ZEA and GLUS by Pediococcus acidilactici H47 increased, among which the average degradation rate of AFB1, DON, ZEA and GLUS could reach 62.74% at 14 days of fermentation.

[0143] The Pediococcus acidilactici H47 provided by the present invention has a deposit number of CCTCCNO: M20232100 and was deposited in the China Center for Type Culture Collection on November 1, 2023. The strain is isolated from the rumen of Qinghai Tibetan sheep.

[0144] Pediococcus acidilactici H47 has stronger environmental adaptability and stronger vitality; it has high tolerance to acid and bile salts; it has no hemolytic symptoms; it has strong inhibitory effects on Salmonella and Escherichia coli, and extremely strong inhibition on Staphylococcus aureus. It can efficiently degrade AFB1, DON, ZEA and GLUS, and can efficiently degrade mycotoxins, indicating that this strain is not only safe, but also has good probiotic properties. It can be used as a feed additive or as a strain to ferment feed, degrading mycotoxins in feed while improving feed utilization.

[0145] The present invention separates and screens lactic acid bacteria that can efficiently degrade mycotoxins and have probiotic properties from the rumen of Qinghai Tibetan sheep. The lactic acid bacteria ferment feed, which can not only degrade multiple fungal toxins in feed raw materials and improve the safety of feed, but also increase the utilization of some unconventional feed raw materials. The invention has a very good prospect in the application of fermented feed with unconventional feed as raw material.

[0146] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various changes can be made to the technical scheme of the present invention. These simple variations all belong to the protection scope of the present invention.

[0147] It should also be noted that the various specific technical features and steps described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

[0148] In addition, various embodiments of the present invention may be arbitrarily combined, and as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

Claims

1. A Pediococcus acidilactici, characterized in that: The Pediococcus acidilactici is Pediococcus acidilactici H47, the deposit number is CCTCC NO: M20232100, and it is deposited in China Center for Type Culture Collection.

2. A microbial preparation, characterized in that: The microbial preparation comprises the Pediococcus acidilactici according to claim 1 Pediococcus acidilactici H47 or cultures of said strains.

3. A composition, characterized in that: The composition comprises the Pediococcus acidilactici according to claim 1 Pediococcus acidilactici H47 and at least one of a culture of said strain; The culture of the strain includes the Pediococcus acidilactici Pediococcus acidilactici Fermentation broth, fermentation precipitate or lyophilized powder of H47.

4. The composition according to claim 3, characterized in that: The composition is a microecological preparation or a pharmaceutical preparation.

5. The composition according to claim 4, characterized in that: The composition is a probiotic.

6. The composition according to claim 3, characterized in that: The composition further comprises a substance which helps to maintain the vitality of Pediococcus acidilactici; the composition further comprises other probiotics and / or prebiotics.

7. Use of the Pediococcus acidilactici according to claim 1, or the microbial preparation according to claim 2, or the composition according to any one of claims 3 to 6 in the preparation of a composition for degrading aflatoxin B1, vomitoxin, zearalenone, and glucosinolates.

8. Use of the Pediococcus acidilactici according to claim 1, or the microbial preparation according to claim 2, or the composition according to any one of claims 3 to 6 in the preparation of a composition for a probiotic.

9. Use of the Pediococcus acidilactici according to claim 1, or the microbial preparation according to claim 2, or the composition according to any one of claims 3 to 6 in preparing a composition for feed fermentation.

10. An animal feed, characterized in that: The animal feed comprises the Pediococcus acidilactici and / or a fermentation product of Pediococcus acidilactici according to claim 1 and / or the microbial preparation according to claim 2 and / or the composition according to any one of claims 3 to 6.

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