CN113736668B - Aspergillus niger strain and application thereof in ochratoxin A degradation - Google Patents
Aspergillus niger strain and application thereof in ochratoxin A degradation Download PDFInfo
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Abstract
The invention discloses an aspergillus niger strain and application thereof in ochratoxin A degradation, and belongs to the field of genetic engineering. The invention provides an aspergillus niger (Aspergillus niger) FS-UV-21 which is preserved in China center for type culture collection of microorganisms (CGMCC) No.20751 in the year 11 and 9 of 2020. Aspergillus niger (Aspergillus niger) FS-UV-21 has good degradation effect on ochratoxin A, the degradation rate reaches 74.49%, and the degradation rate is improved by 18.23% compared with the original strain; the time for reaching the optimal degradation effect is advanced, and 18 hours is shortened; aspergillus niger belongs to food-grade microorganisms, has higher food safety, and can realize the biological high-efficiency degradation of mycotoxin.
Description
Technical Field
The invention relates to an aspergillus niger strain and application thereof in ochratoxin A degradation, belonging to the field of genetic engineering.
Background
Mycotoxins are secondary metabolites produced by fungi, which are widely found in agricultural products and their preparations, and mycotoxin contamination has become one of the major global food-borne risks facing human health due to serious harm to humans and animals. More than 400 eubacterial toxins, which have been discovered by scientists at present, are toxic and have carcinogenic, mutagenic and other properties although they all exhibit different physiological and biochemical properties, and it is known that disorders of the human and animal immune system have a certain relationship with mycotoxins. The most common mycotoxins at present include ochratoxins, aflatoxins, zearalenol, vomitoxins and the like, which are the mycotoxins currently found to be the most harmful to grains, feeds, animals and humans. Ochratoxins, in particular ochratoxin a (OTA), are considered one of the five most important mycotoxins that contaminate foods and feeds. OTA has nephrotoxicity, hepatotoxicity, teratogenicity, immunotoxicity, and genotoxicity, etc., and has been classified by the international cancer research institution as a class 2B carcinogen. OTA is a great hazard and there is a need to find an effective and safe technique to reduce ochratoxin levels to a threshold.
The microbial control has the advantages of safety, high efficiency, economy, strong applicability, environmental friendliness and the like, and becomes a control method which is favored in recent years. Research shows that a variety of microorganisms have a removal effect on OTA, including lactobacillus, acinetobacter, yeast, rhizopus, aspergillus and the like. Zhou Yo, et al, show that Acinetobacter calcoaceticus and Acinetobacter calcoaceticus in soil have the ability to degrade OTA (see Zhou Yo, ji Xiaofeng, li Wenjun.2012. Aspergillus mycotoxin pollution, hazard and biological detoxification techniques, innovative [ J ]. Proc. Of Chinese veterinarian, 32 (11): 1741-1746); liu Changyu A strain of Garcinia having the function of double degradation of OTA and AFB1, which can effectively degrade OTA and AFB1 under the condition of low concentration of toxin pollution (see Liu Changyu. The degradation characteristics of Garcinia Cambogia CW574 on AFB_1 and OTA and the application thereof in biological detoxification [ D ]. Anhui agricultural university, 2019.); research on the pandas and the like shows that the Aspergillus niger M00988 strain has the effect of degrading OTA (see the pandas, zhi Huiwei, wang Xiaoyi and the like; prokaryotic expression of ochratoxin A degrading carboxypeptidase genes and immobilization thereof from the Aspergillus niger M00988 strain [ J ]. J. Chinese food journal, 2019,019 (005): 66-75). The strain capable of degrading OTA is various, but the strain which has good effect and high safety and can be effectively applied to the degradation of toxin in actual samples is less. In addition, strains that degrade multiple mycotoxins are of greater utility than strains that degrade a single toxin. Therefore, the separated strain has very important significance for preventing and curing mycotoxins such as ochratoxin and the like and degrading effect.
Previous studies have shown that A.niger has a better degradation of OTA toxins. However, the Aspergillus niger strains currently used for degrading OTA are less effective and take longer. For example, aspergillus niger W35 disclosed in patent CN 107312719B has a 48h degradation rate of 56.0% at pH 6.5, but the fermentation broth is prepared for a longer period of time. In addition, the existing aspergillus niger only has a single action, namely only can act on mycotoxin OTA singly, and in the practical application environment, the existing aspergillus niger can be a mixed environment of various mycotoxins, such as: ochratoxin a, aflatoxin B1 (AFB 1), zearalenone (ZEN) and vomitoxin (DON). In addition, many studies have only stopped degrading in the medium, lacking the biodegradation effect in the actual sample. Therefore, the aspergillus niger capable of efficiently degrading various mycotoxins can have good application prospect in industry.
Disclosure of Invention
In order to solve the problem that a strain which can degrade various mycotoxins simultaneously and can be applied to the degradation of ochratoxin A in grains is lacking in the prior art, the invention provides an aspergillus niger (Aspergillus niger) FS-UV-21 which is preserved in China center for type culture collection of microorganisms (CGMCC No. 20751) on the 11 th month 9 of 2020.
The Aspergillus niger FS-UV-21 is obtained by carrying out mutagenesis on Aspergillus niger FS-Z1 by adopting an ultraviolet irradiation mode, repeating 5 times, selecting 40 strains after irradiation, comparing degradation rates of each strain on OTA, selecting the strain with the highest degradation rate for preservation and identification, and combining morphological characteristics and molecular biological identification of the strain to determine that the strain is Aspergillus niger. Phylogenetic tree based on its 18S rDNA full sequence (SEQ NO. ID 1) was designated A.niger (Aspergillus niger) FS-UV-21 and sent to the collection for preservation.
The Aspergillus niger FS-Z1 is described in the Chinese patent application text with publication number CN 103937681B.
The Aspergillus niger has the following preservation conditions: inoculating Aspergillus niger with 3 rings of mutagenesis on a solid plate culture medium PDA with good growth, culturing for 36h at 28deg.C in a shaking table (180 rpm), transferring 0.45mL into glycerol pipe containing 0.45mL of sterile glycerol, and storing in a-80deg.C ultra-low temperature refrigerator.
The solid state plate medium PDA was (w/v): 300g of potato, 20g of glucose, 20g of agar, 0.1g of chloramphenicol, 1L of distilled water, natural pH and high pressure sterilization at 121 ℃ for 15min.
The liquid fermentation medium PDB is (w/v): 300g of potato, 20g of glucose, 1L of distilled water, and sterilizing at a natural pH of 121 ℃ for 15min. The culture conditions of the mutagenized aspergillus niger are as follows: the mutagenesis Aspergillus niger with preserved glycerol is inoculated into a liquid culture medium, and is subjected to shaking culture for 48 hours at 28 ℃ and 180 rpm.
The invention also provides a microbial preparation containing the aspergillus niger (Aspergillus niger) FS-UV-21.
In one embodiment of the present invention, the concentration of Aspergillus niger (Aspergillus niger) FS-UV-21 in the microbial preparation is at least 1X 10 6 CFU/mL。
In one embodiment of the invention, the microbial agent is a solid, liquid microbial agent or a lyophilized powder.
The invention also provides a product, which contains the aspergillus niger (Aspergillus niger) FS-UV-21 or the microbial preparation.
In one embodiment of the invention, the concentration of Aspergillus niger (Aspergillus niger) FS-UV-21 in the product is at least 1X 10 6 CFU/mL。
In one embodiment of the invention, the product is a feed additive or chemical.
The invention also provides application of the aspergillus niger (Aspergillus niger) FS-UV-21, or the microbial preparation, or the product in degrading mycomycin.
In one embodiment of the invention, the mycotoxins include one or more of ochratoxin a, aflatoxin B1, zearalenone, vomitoxin.
The invention also provides a method for simultaneously degrading ochratoxin A (OTA), aflatoxin B1 (AFB 1), zearalenone (ZEN) and vomitoxin (DON), which comprises the step of adding the aspergillus niger (Aspergillus niger) FS-UV-21 or the microbial preparation or the product into an environment containing the aflatoxin B1, zearalenone, ochratoxin A and vomitoxin for degradation.
In one embodiment of the invention, the aspergillus niger (Aspergillus niger) FS-UV-21 is added in an amount of at least: 1X 10 6 CFU/mL。
In one embodiment of the invention, the Aspergillus niger (Aspergillus niger) FS-UV-21 is added to the reaction system in the form of a bacterial suspension that is inoculated into the reaction system in an inoculum size of 2% (v/v).
In one embodiment of the invention, the degradation conditions are: aspergillus niger spore concentration was 1X 10 6 CFU/mL, the addition amount is 2% (v/v), the content of OTA, AFB1, ZEN and DON in the PDB liquid culture medium is 1ppm respectively, and the reaction conditions are as follows: shaking culture is carried out for 48h at 28 ℃ and 180 rpm.
The invention also provides a method for degrading ochratoxin A, which comprises the step of adding the aspergillus niger (Aspergillus niger) FS-UV-21 or the microbial preparation or the product into an environment containing the ochratoxin A for degradation.
In one embodiment of the invention, the aspergillus niger (Aspergillus niger) FS-UV-21 is added in an amount of at least: 2% (v/v).
In one implementation of the inventionIn a mode, the degradation conditions are as follows: aspergillus niger spore concentration was 1X 10 6 CFU/mL, the addition amount is 2% (v/v), the OTA content in the PDB liquid culture medium is 1ppm, and the reaction conditions are as follows: shaking culture is carried out for 48h at 28 ℃ and 180 rpm.
The invention also provides application of the aspergillus niger (Aspergillus niger) FS-UV-21 or the microbial preparation in preparation of degradable mycotoxin products.
In one embodiment of the invention, the product is a chemical or feed additive.
In one embodiment of the invention, the mycotoxin is one or more of ochratoxin a (OTA), aflatoxin B1 (AFB 1), zearalenone (ZEN), vomitoxin (DON).
In one embodiment of the invention, the aspergillus niger (Aspergillus niger) FS-UV-21 is added in an amount of at least: 1X 10 6 CFU/mL。
Advantageous effects
(1) Aspergillus niger (Aspergillus niger) FS-UV-21 has good degradation effect on ochratoxin A, the degradation rate reaches 74.49%, and the degradation rate is improved by 18.23% compared with the original strain; the time for reaching the optimal degradation effect is advanced and shortened by 18 hours.
(2) Aspergillus niger (Aspergillus niger) FS-UV-21 has good passage stability.
(3) Aspergillus niger (Aspergillus niger) FS-UV-21 has good degradation effect in the environment containing ochratoxin A, aflatoxin B1, zearalenone and vomitoxin, and the degradation rates are 66.09%, 74.61%, 88.29% and 37.76% respectively.
(4) Aspergillus niger (Aspergillus niger) FS-UV-21 is used for removing wheat bran OTA toxin in a biological fermentation mode, and the removal rate is 59.74% at the highest.
(5) Aspergillus niger belongs to food-grade microorganisms, has higher food safety, and can realize the biological high-efficiency degradation of mycotoxin.
Preservation of biological materials
A strain of Aspergillus niger (Aspergillus niger) FS-UV-21 is named as Aspergillus niger Aspergillus niger, and is preserved in China general microbiological culture Collection center (CGMCC) No.20751 at 11/09/2020, and has a preservation address of Beijing Kogyo-region North Chen West Lu No. 1/3/China academy of sciences microbiological culture Collection center.
Drawings
Fig. 1: the degradation effect of the mutagenized Aspergillus niger strain on OTA.
Fig. 2: phylogenetic tree of A.niger (Aspergillus niger) FS-UV-21.
Fig. 3: the degradation effect of the original strains Aspergillus niger (Aspergillus niger) FS-Z1 and Aspergillus niger (Aspergillus niger) FS-UV-21 on OTA is dynamically changed.
Fig. 4: passage stability of A.niger (Aspergillus niger) FS-UV 21.
Fig. 5: aspergillus niger (Aspergillus niger) FS-UV21 degrades multiple toxins simultaneously.
Fig. 6: aspergillus niger (Aspergillus niger) FS-UV21 removes OTA from wheat bran.
Detailed Description
The starting strain Aspergillus niger FS-Z1 in the examples below is described in the Chinese patent publication No. CN 103937681B.
The following examples relate to the following media:
PDA solid medium: 300g of potato, 20g of glucose, 20g of agar, 0.1g of chloramphenicol, 1L of distilled water, natural pH and high pressure sterilization at 121 ℃ for 15min.
PDB liquid fermentation medium: 300g of potato, 20g of glucose, 1L of distilled water, and sterilizing at a natural pH of 121 ℃ for 15min.
The detection method involved in the following examples is as follows:
extraction of ochratoxin a (OTA) in liquid medium:
1mL of the liquid to be tested is taken, 3mL of chloroform is added, the mixture is fully oscillated, the mixture is centrifuged at 5000rpm for 10min, the lower chloroform layer is taken out, and the mixture is transferred to a new centrifuge tube. Repeating the extraction twice, vacuum centrifuging, freezing and volatilizing the extract, adding 1mL of acetonitrile-2% acetic acid for water dissolution, filtering, and storing at 4 ℃ for high performance liquid chromatography detection.
Extraction of ochratoxin a (OTA) from wheat bran:
1g of wheat bran is taken, 10mL of 60% acetonitrile water solution is added, vortex oscillation is carried out for 30s, ultrasound is carried out for 30min, centrifugation is carried out at 8000rpm for 10min, supernatant is taken, vacuum centrifugation and freeze-drying are carried out, 1mL of acetonitrile-2% acetic acid water solution is added for dissolution, and after filtration, storage is carried out at 4 ℃ for high performance liquid chromatography detection.
High performance liquid chromatography (hplc) to detect ochratoxin a (OTA) conditions: high performance liquid chromatography species: agilent Technologies 1260 index; chromatographic column: c18 column, column length: 150mm, inner diameter: 4.6mm, viscosity: 3 μm; mobile phase: acetonitrile: water: acetic acid=96:102:2; flow rate: 1.0mL/min; detection wavelength: excitation light wavelength: 333nm, emission wavelength: 460nm.
Extraction and detection of aflatoxin B1 (AFB 1) in liquid medium:
sucking 1ml of the solution to be detected, adding 3ml of chloroform, extracting twice, vacuum centrifuging, freezing, volatilizing, re-dissolving 20% acetonitrile, adding 100ul of trifluoroacetic acid and 200ul of n-hexane, derivatizing for 30min, and detecting by using a high performance liquid chromatography. The chromatographic conditions were C18 column 6 mm. Times.150 mm. Times.5 um; mobile phase acetonitrile: water = 20:80 detection temperature: 30 ℃; flow rate: 1ml/min; detection wavelength: excitation wavelength: 360nm; emission wavelength: 440nm.
Extraction and detection of Zearalenone (ZEN) in liquid medium:
sucking 1ml from the solution to be detected, adding 3ml of chloroform, extracting twice, vacuum centrifuging, freezing, volatilizing, redissolving 50% acetonitrile, and performing content detection by high performance liquid chromatography. The chromatographic conditions are chromatographic columns: c18 column 6 mm. Times.150 mm. Times.5 um; mobile phase: acetonitrile: water=50:50; detecting the temperature: 25 ℃; flow rate: 1ml/min; detection wavelength: excitation wavelength: 240nm; emission wavelength: 440nm.
Extraction and detection of vomitoxin (DON) in liquid medium:
extracting with 3ml chloroform twice by sucking 1ml, vacuum centrifuging, freeze volatilizing, redissolving with 20% methanol, and detecting content by high performance liquid chromatography. Chromatographic conditions: the chromatographic conditions are chromatographic columns: c18 column 6 mm. Times.150 mm. Times.5 um; mobile phase: methanol: water = 20:80; detecting the temperature: 30 ℃; flow rate: 0.8ml/min; detection wavelength: 218nm.
The method for calculating the degradation rate of ochratoxin A (OTA), aflatoxin B1 (AFB 1), zearalenone (ZEN) and vomitoxin (DON) comprises the following steps: (1000-residual concentration)/1000×100%.
Example 1: preparation of Aspergillus niger (Aspergillus niger) FS-UV-21
The method comprises the following specific steps:
and (3) carrying out mutagenesis on the aspergillus niger FS-Z1 by adopting an ultraviolet irradiation mode, repeating 5 times, selecting 40 strains after irradiation, comparing degradation rates of each strain on OTA, and selecting the strain with the highest degradation rate for preservation and identification.
The specific mutagenesis steps are as follows:
1. inoculating Aspergillus niger parent selected from soy sauce grains into PDA solid culture medium, culturing at 28deg.C for 120 hr, collecting 50mL physiological saline, washing spores, shaking uniformly to form monospore suspension, and making into 1×10 6 CFU/mL of bacterial suspension.
2. Taking 5mL of the spore suspension, and repeating for 5 times by irradiating with 200W ultraviolet for 30 min.
3. 40 strains after mutagenesis are selected, the serial numbers are 1 to 40 respectively, the 40 strains are inoculated in PDB solid culture medium, proper spores are selected from bacterial colonies, and 10 is prepared according to the mode 6 CFU/mL of the bacterial suspension, and the bacterial suspension was inoculated into 50mL of a medium containing 1ppm of OTA at 2% (v/v) and shake-cultured at 28℃at 180rpm for 48 hours.
4. The degradation effect of the original strain and the novel mutant strain on OTA is compared (the result is shown in figure 1), and the strain FS-UV-21 with the highest degradation efficiency is selected and stored.
By colony characterization, individual morphology was observed and 18S rDNA of strain FS-UV-21 (sequence shown as SEQ NO. ID 1) was compared to known sequences in the GenBank database and found to be 99% homologous to A.niger (Aspergillus niger).
The strain can be determined to be Aspergillus niger by combining morphological characteristics and molecular biological identification of the strain. Phylogenetic tree based on its 18S rDNA full sequence is shown in figure 2 of the specification. As can be seen from FIG. 1, strain FS-UV-21 has a highest degradation rate of 74.49% to OTA, and is designated as Aspergillus niger FS-UV-21 and sent to the collection center for preservation.
Example 2: dynamic changes in A.niger (Aspergillus niger) FS-UV-21 degradation OTA
The method comprises the following specific steps:
1. inoculating Aspergillus niger FS-UV-21 and Aspergillus niger FS-Z1 into PDA solid culture medium, culturing at 28deg.C for 120 hr, washing spores with 50mL physiological saline containing 0.02% (v/v) Tween 80, shaking uniformly to form monospore suspension, wherein the concentration of the spore suspension is: 1X 10 6 CFU/mL。
2. The spore suspensions were inoculated into 50mL of PDB medium containing 1ppm of OTA at an inoculum size of 2% (v/v), shaking culture was carried out at 28℃and 180rpm for 72 hours, and the corresponding samples were extracted every 6 hours to detect changes in OTA concentration, and the results are shown in Table 1 and FIG. 3.
Table 1: degradation Rate of OTA at different reaction times (%)
As can be seen from fig. 3, the degradation efficiency is significantly improved relative to the original strain; the time for achieving the optimal degradation effect is shortened from 60 hours to 42 hours, which shortens 18 hours and can degrade mycotoxin more quickly and efficiently.
Example 3: passage stability of Aspergillus niger FS-UV-21
The method comprises the following specific steps:
1. inoculating Aspergillus niger FS-UV-21 into PDA solid culture medium, and passaging for 5 generations, wherein the method comprises the following steps: the strain of the first generation is transferred to a new PDA solid culture medium after 7d of culture at 28 ℃, the strain of the first generation is transferred to a new PDA solid culture medium after 7d of culture at 28 ℃ to be the second generation, and the operation is repeated to the fifth generation, so that degradation effects are detected respectively.
2. According to the method of example 2 (bacterial concentration 10 6 CFU/mL, OTA concentration of 1 ppm), treating the OTA-containing solution with Aspergillus niger FS-UV-21 for 48h, and detecting the determination reaction of the Aspergillus niger FS-UV-21 on OTA after each passageIs a degradation rate of (a).
As can be seen from FIG. 4, the degradation rate of the Aspergillus niger after 5 passages can be stabilized at about 75%, and the genetic stability is good.
Example 4: aspergillus niger FS-UV-21 simultaneously degrades ochratoxin A, aflatoxin B1, zearalenone and vomitoxin
The method comprises the following specific steps:
1. inoculating Aspergillus niger FS-UV-21 into PDA solid culture medium, culturing at 28deg.C for 120 hr, taking 50mL physiological saline containing 0.02% (v/v) Tween 80 to wash out spores, shaking uniformly to form monospore suspension, wherein the concentration of the spore suspension is: 1X 10 6 CFU/mL。
2. The spore suspensions were inoculated into 50mL PDB medium containing 1ppm OTA, 1ppm AFB1, 1ppm ZEN and 1ppm DON according to an inoculum size of 2% (v/v), and subjected to shaking culture at 28℃at 180rpm for 42 hours, and the degradation rates of the different toxins after the completion of the reaction were determined.
As shown in FIG. 5, the degradation rates of ochratoxin A, aflatoxin B1, zearalenone and vomitoxin are 66.09%, 74.61%, 88.29% and 37.76%, respectively, and the Aspergillus niger FS-UV-21 can degrade various mycotoxins simultaneously with good degradation effect.
Example 5: degradation of wheat bran by Aspergillus niger FS-UV-21
The method comprises the following specific steps:
1. inoculating Aspergillus niger FS-UV-21 into PDA solid culture medium, culturing at 28deg.C for 120 hr, taking 50mL physiological saline containing 0.02% (v/v) Tween 80 to wash out spores, shaking uniformly to form monospore suspension, wherein the concentration of the spore suspension is: 1X 10 6 CFU/mL。
2. Treating wheat bran: crushing wheat bran, sieving with a 40-mesh sieve, uniformly mixing and collecting the crushed wheat bran, and adding an OTA standard substance into the wheat bran to ensure that the OTA content in the wheat bran is 1 mug/g.
3. Weighing 10g of the wheat bran treated in the step 2, subpackaging in 50mL triangular conical flasks, sealing four layers of gauze newspaper, and sterilizing at 121 ℃ for 20 min.
4. Transferring the wheat bran subjected to sterilization treatment obtained in the step 3 into a fermentation container, adding sterile water to a ratio of 1:3 (the ratio of the mass of the wheat bran to the volume of the sterile water, g/mL), adding the FS-UV-21 spore suspension prepared in the step 1 according to the inoculation amount of 15% of the mass of the wheat bran and the volume of the bacterial liquid, uniformly stirring, performing constant-temperature fermentation at 28 ℃, sampling after fermentation for 1d, 2d, 3d, 4d and 5d, detecting the residual amount of OTA, and calculating the degradation rate, wherein the results are shown in Table 2 and FIG. 6.
Table 2: degradation rate of OTA in wheat bran by Aspergillus niger with different fermentation time
As can be seen from FIG. 6, different fermentation times have an important effect on the removal of OTA from wheat bran by Aspergillus niger, and the degradation rate of OTA increases with increasing fermentation time. At fermentation time 5d, the degradation rate is 59.74% at the highest.
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
SEQUENCE LISTING
<110> university of Jiangnan
<120> A strain of Aspergillus niger and its use in the degradation of ochratoxin A
<130> BAA211120A
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 619
<212> DNA
<213> artificial sequence
<400> 1
ggcggccggt tcctccgcct tttgatatgc ttaagttcag cgggtatccc tacctgatcc 60
gaggtcaacc tggaaagaat ggttggaaaa cgtcggcagg cgccggccaa tcctacagag 120
catgtgacaa agccccatac gctcgaggat cggacgcggt gccgccgctg cctttcgggc 180
ccgtcccccc ggagaggggg acggcgaccc aacacacaag ccgggcttga gggcagcaat 240
gacgctcgga caggcatgcc ccccggaata ccagggggcg caatgtgcgt tcaaagactc 300
gatgattcac tgaattctgc aattcacatt agttatcgca tttcgctgcg ttcttcatcg 360
atgccggaac caagagatcc attgttgaaa gttttaactg attgcattca atcaactcag 420
actgcacgct ttcagacagt gttcgtgttg gggtctccgg cgggcacggg cccggggggc 480
agaggcgccc ccccggcggc cgacaagcgg cgggcccgcc gaagcaacag ggtacaatag 540
acacggatgg gaggttgggc ccaaaggacc cgcactcggt aatgatcctt ccgcaggtcc 600
ccctaacgga agggcggta 619
Claims (8)
1. Aspergillus niger strainAspergillus niger) The strain is preserved in China general microbiological culture collection center (CGMCC) No.20751 at 11 and 09 of 2020.
2. A microbial preparation comprising aspergillus niger according to claim 1.
3. The microbial preparation according to claim 2, wherein the content of aspergillus niger in the microbial preparation is at least 1 x 10 6 CFU/mL。
4. A product comprising aspergillus niger according to claim 1 or a microbial preparation according to claim 2 or 3, said product being a feed additive.
5. The product of claim 4, wherein the Aspergillus niger content of the product is at least 1X 10 6 CFU/mL。
6. Use of an aspergillus niger according to claim 1, or a microbial preparation according to claim 2 or 3, or a product according to claim 4 or 5, for degrading mycotoxins, which are one or more of ochratoxin a, aflatoxin B1, zearalenone, vomitoxin.
7. A method of degrading ochratoxin a, characterized in that aspergillus niger according to claim 1, or a microbial preparation according to claim 2 or 3, or a product according to claim 4 or 5, is added to an environment containing ochratoxin a for degradation, said method being for non-diagnostic therapeutic purposes.
8. Use of an aspergillus niger according to claim 1, or a microbial preparation according to claim 2 or 3, for the preparation of a degradable mycotoxin product, characterized in that the product is a feed additive and the mycotoxin is one or more of ochratoxin a, aflatoxin B1, zearalenone, vomitoxin.
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