CN108251323B - Bacillus licheniformis, microbial inoculum containing bacillus licheniformis, application of microbial inoculum, method for degrading vomitoxin and kit - Google Patents
Bacillus licheniformis, microbial inoculum containing bacillus licheniformis, application of microbial inoculum, method for degrading vomitoxin and kit Download PDFInfo
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Abstract
The invention relates to the field of microorganisms, and discloses bacillus licheniformis, a microbial inoculum containing the bacillus licheniformis, application of the bacillus licheniformis, a method for degrading vomitoxin and a kit. Specifically, the invention provides a Bacillus licheniformis (Bacillus licheniformis), and the preservation number of the Bacillus licheniformis is CGMCC NO. 13314. The bacillus licheniformis provided by the invention can efficiently and quickly degrade vomitoxin in grain oil and/or feed, and particularly, even if the content of the vomitoxin in the grain oil and/or feed is more than 50ppm, the bacillus licheniformis can efficiently and quickly degrade the vomitoxin and has no influence on the palatability of the grain oil and/or feed; meanwhile, no toxic product is generated when the bacillus licheniformis is used for degrading vomitoxin, and the bacillus licheniformis is safe and environment-friendly. Therefore, the bacillus licheniformis has good application prospect.
Description
Technical Field
The invention relates to the field of microorganisms, in particular to bacillus licheniformis, a microbial inoculum containing the bacillus licheniformis, application of the bacillus licheniformis, a method for degrading vomitoxin and a kit.
Background
Vomitoxin (vomitoxin), also known as Deoxynivalenol (DON), is named after 3 α,7 α, 15-trihydroxy Fusarium solani-9-en-8-one, because it can cause vomiting of pigs, is mainly trichothecene toxins produced by Fusarium graminearum (Fusarium graminearum) and Fusarium yellow (Fusarium culmorum) infecting grains such as wheat, barley, oats and corn. Vomitoxin is one of main mycotoxin pollution of grains, feeds and foods worldwide, and seriously affects the health of people and livestock. After people and livestock ingest food polluted by vomitoxin, acute poisoning symptoms such as anorexia, vomit, diarrhea, fever, unstable standing, slow response and the like can be caused, the hematopoietic system is damaged in serious cases to cause death, and the serious harmfulness of the food has attracted general attention of various countries.
The vomitoxin has quite common pollution to grain raw materials in China, the detection rate and the detection amount of the vomitoxin are the highest one of mycotoxins, and investigation by screening sunshine and the like shows that the standard exceeding proportion of the vomitoxin of Chinese feed and raw materials is close to 70%, the standard exceeding rate of the vomitoxin in corn is 57.1%, the average content of the toxin is 1.01mg/kg, and the highest content of the vomitoxin is 2.13 mg/kg. Because of the ubiquitous nature, high content and acute and chronic toxicity of vomitoxin in grains and feeds, it is important and urgent to reduce or eliminate the toxicity. At present, the detoxification method of vomitoxin at home and abroad mainly comprises three major methods, namely a physical method, a chemical treatment and a biological method. Although detoxification by physical and chemical methods has been successful to a certain extent, the disadvantages of limited detoxification effect, possible loss of important nutrients, high cost and the like still exist, so that the application of the two methods in actual production is limited.
The biological method mainly utilizes microorganisms or degradation products thereof to carry out toxin degradation, has the advantages of reducing toxicity of toxins under mild conditions, having small influence on sensory properties, palatability and nutrient substances of raw materials and the like, and has the characteristics of safety, environmental protection and high efficiency, thereby being considered as the optimal detoxification method. Therefore, the research of removing vomitoxin in grain and oil or/feed by using modern biotechnology has good application prospect. The patent application CN103243047A discloses a bacillus subtilis for efficiently degrading vomitoxin and an application thereof, wherein 900 mu L of bacillus subtilis ANSB471 fermentation liquor reacts with 100 mu L of vomitoxin (100 mu g/ml), the degradation rate of the vomitoxin is 25% after 2 hours of reaction, the degradation rate of the vomitoxin is 56% after 24 hours of reaction, the degradation rate of the vomitoxin is 80% after 48 hours of reaction, and the degradation rate is yet to be improved.
In addition, the existing biological methods for degrading vomitoxin are mostly carried out under mild conditions (such as temperature 25-37 ℃ and not more than 40 ℃ at the maximum, and pH value is about 7), however, no good solution exists under higher temperature load (such as the condition of transportation in a container or during feed granulation) or under severe acid-base conditions, which limits the application range of the biological methods in degrading the vomitoxin.
Therefore, there is a need to find a method for biodegrading vomitoxin that is efficient and safe and can be used under high temperature load or harsh acid-base conditions.
Disclosure of Invention
The invention aims to overcome the defects in the existing vomitoxin degradation process, and provides a bacillus licheniformis, a microbial inoculum containing the bacillus licheniformis, application of the bacillus licheniformis, a method for degrading vomitoxin and a kit.
In order to achieve the above object, in a first aspect, the present invention provides a strain of Bacillus licheniformis (Bacillus licheniformis), wherein the preservation number of the Bacillus licheniformis is CGMCC No. 13314.
In a second aspect, the present invention also provides a microbial inoculum, wherein the microbial inoculum contains the Bacillus licheniformis (Bacillus licheniformis).
In a third aspect, the invention also provides application of the bacillus licheniformis and/or the microbial inoculum in degrading vomitoxin.
In a fourth aspect, the present invention also provides a method for degrading emetic toxin, wherein the method comprises: and (3) contacting the bacillus licheniformis and/or the microbial inoculum with a vomitoxin-polluted sample to degrade the vomitoxin in the sample.
In a fifth aspect, the present invention provides a kit comprising the bacillus subtilis and/or the microbial inoculum of the present invention.
The bacillus licheniformis provided by the invention can effectively and quickly degrade vomitoxin in grain oil and/or feed, and particularly, even if the content of the vomitoxin in the grain oil and/or feed is more than 50ppm (preferably at least 100ppm, more preferably at least 150ppm, and even more preferably at least 200ppm), the bacillus licheniformis can effectively and quickly degrade the vomitoxin and has no influence on the palatability of the grain oil and/or feed (namely, the taste is basically not different before and after treatment); meanwhile, the bacillus licheniformis is added into grain oil and/or feed polluted by vomitoxin, no toxic product is generated, and the safety is realized. Therefore, the bacillus licheniformis has good application prospect.
In addition, the bacillus licheniformis provided by the invention also has high temperature and acid-base stability, and particularly has good stability under alkaline conditions (such as pH of 8-10), so that the application range of the bacillus licheniformis is further expanded.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Biological preservation
Bacillus licheniformis (Bacillus licheniformis) is preserved in the China general microbiological culture Collection center (address: No. 3 of West Lu No.1 of North Chen of the Korean district, Beijing, China academy of sciences, microbiological research institute, postal code: 100101) (the abbreviation of the preservation unit is CGMCC) in 2016 (11.16.16.11.15.14.M.) with the preservation number of CGMCC NO. 13314.
Detailed Description
The following describes in detail specific embodiments of the present invention. It should be understood that the detailed description and specific examples, while indicating the present invention, are given by way of illustration and explanation only, not limitation.
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.
In a first aspect, the invention provides a Bacillus licheniformis (Bacillus licheniformis), wherein the preservation number of the Bacillus licheniformis is CGMCC NO. 13314.
The bacillus licheniformis provided by the invention is separated from a soil sample (collected in Beijing) polluted by vomitoxin. The isolation of the Bacillus licheniformis can be carried out by methods conventional in the art for isolation of new strains, such as liquid phase enrichment or soil circulation.
The liquid phase enrichment method may specifically include: weighing a proper amount of soil sample polluted by vomitoxin, adding the soil sample into a triangular flask filled with LB liquid culture medium, and adding a proper amount of glass beads. Oscillating the triangular flask at 28-37 ℃ and 160-; transferring the soil mixed solution into a centrifuge tube, and taking the centrifuged supernatant as a source of vomitoxin passivation microorganisms. The supernatant was inoculated into LB liquid medium containing vomitoxin and cultured with shaking at 28-37 ℃ and 160-180 rpm. Pipette 1-2mL with a sterile pipette and transfer to another enrichment culture flask. After three such transfers, the supernatants were diluted 10 times each-1、10-2、10-3、10-4、10-5、10-6、10-7Or 10-8Taking out a proper amount of the solution, coating the solution on LB solid culture medium containing 50, 100, 200 or 300mg/L of vomitoxin for plate streaking,after culturing at 28-37 deg.C for 3-4 days, single colony is separated.
The soil circulation method may specifically include: 100g of soil polluted by vomitoxin is weighed and evenly mixed with a proper amount of sand grains with the grain diameter of about 3mm, and the mixture is placed on the upper layer of the circulation device. The lower layer was filled with 200mL of LB medium as a circulating solution. Starting an air compressor, starting the acclimatization process, and periodically supplementing the circulating fluid according to the evaporation condition of the circulating fluid. After the acclimatization is finished, respectively diluting the lower layer circulating fluid by 10-1、10-2、10-3、10-4、10-5、10-6、10-7Or 10-8Taking out appropriate amount of the suspension, spreading the suspension on LB solid medium containing 50, 100, 200 or 300mg/L vomitoxin, streaking, culturing at 28-37 deg.C for 3-4 days, and separating single colony.
The inventor selects a bacterium with the strongest degradation effect on vomitoxin from the screened strains to carry out DNA extraction and identification, and the identification result shows that the 16srDNA sequence of the strain has 100 percent homology with Bacillus licheniformis (Bacillus licheniformis), the strain can be determined to be the Bacillus licheniformis (Bacillus licheniformis), and the strain is preserved in China general microbiological culture Collection center at 2016, 11, 16 and the preservation number is CGMCC NO. 13314.
The bacillus licheniformis provided by the invention can produce a large amount of live thalli of the bacillus licheniformis after being cultured. The culture method of the present invention is not particularly limited as long as the Bacillus licheniformis can be proliferated in a large amount by the culture method, and the amount of Bacillus licheniformis can be 10, for example5Inoculating the live bacteria of the bacillus licheniformis into a culture medium in the inoculation amount of CFU/mL, and culturing at the temperature of 28-38 ℃ for 12-48 hours under aerobic conditions to obtain a culture solution. Wherein the medium may be a medium conventionally used in the art, and for example, may be LB liquid medium (0.8-1 wt% peptone, 0.5-0.8 wt% yeast powder, 1-1.5 wt% sodium chloride; pH 6.8-7.0; culture temperature 28-38 ℃) or nutrient broth medium (0.8-1 wt% peptone, 0.3-0.5 wt% beef extract, 0.5-0.8 wt% sodium chloride; pH 7.2-7.6; culture temperature 28-38℃)) Preferably, LB liquid medium.
The present invention can further separate the thallus of bacillus licheniformis in the culture solution, and the method for separating is not particularly limited as long as the thallus can be enriched from the culture solution, and for example, the method can be realized by centrifugation and/or filtration, and the conditions for centrifugation and filtration can be the conventional conditions in the field, which are well known to those skilled in the art and will not be described herein again.
In a second aspect, the present invention provides a microbial preparation, wherein the microbial preparation comprises the above Bacillus licheniformis (Bacillus licheniformis).
In the present invention, the concentration of bacillus licheniformis in the microbial agent is not particularly limited, and can be specifically selected according to specific situations.
According to the invention, the microbial inoculum contains live bacteria and/or dead bacteria of the bacillus licheniformis. Preferably, the microbial inoculum contains live bacteria or mixed bacteria of live bacteria and dead bacteria of the bacillus licheniformis. When the microbial inoculum contains a mixed biomass of live biomass and dead biomass of the Bacillus licheniformis, the number of the live biomass is preferably higher than that of the dead biomass. Most preferably, the microbial inoculum contains live cells of the bacillus licheniformis.
According to the present invention, the formulation of the microbial inoculum is not particularly limited, and the microbial inoculum can be prepared into different formulations according to different intended uses, and can be added with corresponding components such as excipients, for example, the microbial inoculum can be a liquid microbial inoculum (for example, a bacterial solution) and/or a solid microbial inoculum (for example, a lyophilized microbial inoculum), preferably a solid microbial inoculum. The addition of excipients to the bacterial preparation in any dosage form is well known to those skilled in the art and will not be described in detail herein.
In addition, in the process of research, the inventor of the invention finds that although the bacillus licheniformis provided by the invention is obtained by screening under the condition of high concentration of vomitoxin (at least 50ppm), the bacillus licheniformis has a certain degradation effect on other mycotoxin pollutants (such as vomitoxin, aflatoxin, fumonisin, ochratoxin, T2 toxin and the like) commonly seen in soil.
In a third aspect, the invention also provides an application of the bacillus licheniformis and/or the microbial inoculum in degrading vomitoxin, preferably an application in degrading vomitoxin in grain oil and/or feed.
In a fourth aspect, the present invention also provides a method for degrading emetic toxin, wherein the method comprises: and (3) contacting the bacillus licheniformis and/or the microbial inoculum with a vomitoxin-polluted sample to degrade the vomitoxin in the sample.
According to the invention, the vomitoxin-contaminated sample may be vomitoxin-contaminated grain oil and/or feed.
Preferably, the vomitoxin is present in an amount of at least 1ppm, more preferably at least 50ppm, even more preferably at least 100ppm, even more preferably at least 150ppm, even more preferably at least 200ppm, based on the total weight of the vomitoxin-contaminated sample. In the present invention, "ppm" means "μ g/mL" when the sample to be treated is a liquid; when the sample to be treated is a solid, "ppm" means "μ g/g".
In the present invention, the term "grain and oil" refers to a general term for grains, beans and other grains and oils, and finished products and semi-finished products thereof, and particularly to products that can be eaten by humans. For example, the grain oil may be a grain oil product that is edible to humans and is common in the art, and specifically, the grain oil may include at least one of grains and agricultural byproducts thereof, oil and fat products, wines, milks and products thereof, and the like.
In the present invention, the term "feed" refers to the general term of food for animals raised in agriculture or animal husbandry. For example, the feed may be a food commonly used in the art for feeding animals, and in particular, the feed may include: a) cereals, for example, small grain cereals (such as wheat, barley, rye, oats, and combinations thereof) and/or large grain cereals such as maize or sorghum; b) by-products from cereals, such as corn gluten meal, distillers dried grains with solubles (DDGS), wheat bran, wheat middlings, rice bran, rice hulls, oat hulls, palm kernel, and citrus pulp; c) ensiling the feed; d) proteins from the following sources: such as soy, sunflower, peanut, lupin, pea, broad bean, cotton, canola, fish meal, dried plasma protein, meat and bone meal, potato protein, whey, copra, sesame; e) oils and fats obtained from plant and animal sources; f) minerals and vitamins.
In the present invention, the grain or feed may further comprise a physiologically acceptable carrier, wherein the physiologically acceptable carrier is at least one selected from the group consisting of: maltodextrin, limestone (calcium carbonate), cyclodextrin, wheat bran or wheat component, rice or rice bran, sucrose, starch, Na2SO4And talc and mixtures thereof.
In the present invention, the formulation of the bacillus licheniformis and/or the microbial inoculum is not particularly limited, and the bacillus licheniformis and/or the microbial inoculum can be prepared into different formulations according to different predetermined purposes, and corresponding excipients and other ingredients are added, for example, the bacillus licheniformis and/or the microbial inoculum can be in a liquid state and/or a solid state. Wherein, the addition of excipients to the dosage forms is well known to those skilled in the art, and will not be described in detail herein.
According to the invention, the number of bacteria in the Bacillus licheniformis and/or the microbial inoculum is at least 10 relative to 1g of the sample5CFU, preferably, the number of living bacteria in the bacillus licheniformis and/or the microbial inoculum is at least 105And (4) CFU. In the present invention, the cell count can be measured according to GB 4789.2-94.
According to the present invention, the conditions of the contacting may include: the temperature is 25-55 ℃, the pH value is 5-11, and the time is 1-48 h; preferably, the temperature is 30-40 ℃, the pH value is 7-10, and the time is 12-36 h.
In the present invention, when the sample is a solid, the pH value is determined according to the method of GB/T12456-.
In a fifth aspect, the present invention provides a kit comprising the bacillus subtilis and/or the microbial inoculum of the present invention.
Examples
The present invention will be described in detail below by way of examples.
In the following preparations, preparation comparative examples, examples and comparative examples:
vomitoxin standards were purchased from Sigma, and the remaining experimental materials, unless otherwise specified, were purchased from conventional biochemical stores.
LB liquid medium: 10g of peptone, 5g of yeast powder, 10g of sodium chloride and deionized water are added to the solution to reach a constant volume of 1L, and the pH value is adjusted to 7.
LB solid medium: 10g of peptone, 5g of yeast powder, 10g of sodium chloride, 16g of agar powder and deionized water, wherein the volume is constant to 1L, and the pH value is adjusted to 7.
The strain provided by the invention is Bacillus licheniformis (Bacillus licheniformis), and is preserved in the common microorganism center of China Committee for culture Collection of microorganisms (address: No. 3 of West Lu No.1 of Beijing Kogyo area, morning area, China academy of sciences, microbiological research institute, postal code: 100101) in 2016 (the abbreviation of the preservation unit is CGMCC) in 16 days 11 and 16 days, and the preservation number is CGMCC NO. 13314.
The reference strain 1 is Bacillus licheniformis (Bacillus licheniformis) with the preservation number of CGMCC NO.1.807, purchased from CGMCC.
The reference strain 2 is Bacillus subtilis (CGMCC NO. 7344) published in CN103243047A and purchased from CGMCC.
The cell count was measured according to GB 4789.2-94.
Determining the content of vomitoxin according to the determination immunoaffinity column purification-high performance liquid chromatography of deoxynivalenol in GB/T30956-2014 feed.
The degradation rate (%) of vomitoxin (mass of vomitoxin in the sample before reaction-mass of vomitoxin in the sample after reaction)/mass of vomitoxin in the sample before reaction × 100%.
Preparation example 1
Activating Bacillus licheniformis with preservation number of CGMCC NO.13314 provided by the invention, inoculating the Bacillus licheniformis in an LB liquid culture medium sterilized at 121 ℃ for 15min in an inoculation amount of 1 volume percentCulturing at 160rpm and 37 deg.C under shaking for a time period such that the cell concentration in the bacterial solution is (1 + -0.1) × 105CFU/mL. Simultaneously preparing into dry powder, wherein the total viable count contained in each gram of dry powder is (1 + -0.1) x 105CFU。
Preparation of comparative example 1
According to the method of production example 1, except that Bacillus licheniformis (Bacillus licheniformis) having a preservation number of CGMCC NO.1.807 was used in place of the Bacillus licheniformis having a preservation number of CGMCC NO.13314 provided by the present invention.
Preparation of comparative example 2
According to the method of preparation example 1, except that Bacillus subtilis (Bacillus subtilis) with the preservation number of CGMCC NO.7344 disclosed in CN103243047A is used in place of the Bacillus licheniformis with the preservation number of CGMCC NO.13314 provided by the invention.
Example 1
This example illustrates the ability of bacillus licheniformis provided by the present invention to degrade emetic toxins.
Firstly, measuring degradation time
Putting 900 mu L of the bacterial liquid cultured in the preparation example 1 into a 1.5mL centrifuge tube, adding the vomitoxin standard solution, and uniformly mixing to obtain a mixed solution, wherein the final concentration of the vomitoxin in the mixed solution is 50 ppm.
The mixture was reacted at 37 ℃ and pH 7, and 20. mu.L of the reaction sample was taken for 1 hour, 12 hours, 24 hours, 36 hours, and 48 hours, respectively, to detect the presence of vomitoxin. The results are shown in Table 1.
The results in Table 1 show that the reaction time of 24 hours can achieve a vomitoxin degradation rate of 90% or more. Therefore, 24h was used as the reaction time in the following experiment.
Measurement of thermal stability
Treating the cultured bacterial liquid at 40 deg.C, 45 deg.C, 50 deg.C and 55 deg.C, pH 7 for 30min, respectively placing 900 μ L of the treated bacterial liquid in 1.5mL centrifuge tube, adding vomitoxin standard solution, and mixing to obtain mixed liquid with final concentration of 50ppm of vomitoxin. Then, the mixture was reacted at 37 ℃ and pH 7 for 24 hours, and after completion of the reaction, 20. mu.L of the sample was used for detecting the vomitoxin residue. The results are shown in Table 2.
As can be seen from the results in Table 2, the bacterial solution still has high emetic toxin degradation activity when stored at 40-55 ℃ for 30 min.
③ determination of acid-base stability
Treating the cultured bacterial liquid at pH of 5, 6, 7, 8, 9, 10 and 11 respectively at 37 deg.C for 30min, placing 900 μ L of the treated bacterial liquid in a 1.5mL centrifuge tube, adding vomitoxin standard solution, and mixing to obtain a mixed solution, wherein the final concentration of vomitoxin in the mixed solution is 50 ppm. Then, the mixture was reacted at 37 ℃ and pH 7 for 24 hours, and after completion of the reaction, 20. mu.L of the sample was used for detecting the vomitoxin residue. The results are shown in Table 3.
The results in Table 3 show that the bacterial liquid has high emetic toxin degradation activity even when stored for 30min at a pH of 5-11.
Comparative example 1
The measurement was carried out by the method of example 1, except that the bacterial suspension prepared in preparation example 1 was used instead of the bacterial suspension used in example 1.
The results of the degradation time measurement are shown in table 1, the results of the thermal stability measurement are shown in table 2, and the results of the acid-base stability are shown in table 3.
Comparative example 2
The measurement was carried out by the method of example 1, except that the bacterial suspension prepared in preparation comparative example 2 was used instead of the bacterial suspension used in example 1.
The results of the degradation time measurement are shown in table 1, the results of the thermal stability measurement are shown in table 2, and the results of the acid-base stability are shown in table 3.
TABLE 1
TABLE 2
TABLE 3
Test example 1
10g of the dry powder obtained in preparation example 1 was mixed with 1kg of corn flour contaminated with 200ppm of vomitoxin, 1kg of distilled water was added, and three repetition was performed, and after uniform mixing, detoxification was performed for 1 day at 37 ℃ and pH 7. And then determining the content of vomitoxin in each sample after 1 day of treatment according to the determination immunoaffinity column purification-high performance liquid chromatography of deoxynivalenol in GB/T30956-2014 feed, and calculating the degradation rate. The degradation rate of vomitoxin in the corn flour is calculated to be 90.8%. Also, the addition of dry powder did not have any effect on the palatability of the corn meal.
The results of feeding pigs with the treated corn meal for 14 days show that the diet, drinking water and daily activities of the pigs are normal during the period, and the weight of the pigs always keeps on a stable increasing trend. The above results show that the dry powder obtained in preparation example 1 is added to corn flour for pig feeding, and is safe.
Test example 2
According to the method of test example 1, except that the content of vomitoxin in the corn flour was 100ppm, the degradation rate of vomitoxin was 95.9%. Also, the addition of dry powder did not have any effect on the palatability of the corn meal.
The results of feeding pigs with the treated corn meal for 14 days show that the diet, drinking water and daily activities of the pigs are normal during the period, and the weight of the pigs always keeps on a stable increasing trend. The above results demonstrate the safety of adding dry powder to corn meal for pig feeding.
Test example 3
According to the method of test example 1, except that the content of vomitoxin in the corn flour was 50ppm, the degradation rate of vomitoxin was 100%. Also, the addition of dry powder did not have any effect on the palatability of the corn meal.
The results of feeding pigs with the treated corn meal for 14 days show that the diet, drinking water and daily activities of the pigs are normal during the period, and the weight of the pigs always keeps on a stable increasing trend. The above results demonstrate the safety of adding dry powder to corn meal for pig feeding.
Test comparative example 1
The procedure of test example 1 was followed, except that the dry powder obtained in preparation comparative example 1 was used in place of the dry powder used in example 1. The detected degradation rate of the vomitoxin is 16.8%.
Test comparative example 2
The procedure of test example 1 was followed except that the dry powder obtained in preparation comparative example 2 was used in place of the dry powder used in example 1. The degradation rate of the vomitoxin is 18.1 percent through detection.
Compared with the results of the comparative examples 1-2, the bacillus licheniformis provided by the invention can effectively and rapidly degrade vomitoxin in grain oil and/or feed, and has high temperature and acid-base stability, and particularly has good stability under alkaline conditions (such as pH of 8-10).
As can be seen from the comparison of the results of the above test example 1 and the test comparative examples 1-2, even when the bacillus licheniformis provided by the present invention is applied to grain oil and/or feed with vomitoxin content as high as 50ppm or more, the bacillus licheniformis can effectively and rapidly degrade the vomitoxin and has no influence on the palatability of the grain oil and/or feed (i.e. the taste is not substantially different before and after the treatment); meanwhile, the bacillus licheniformis is added into grain oil and/or feed polluted by vomitoxin, no toxic product is generated, and the safety is realized. Therefore, the bacillus licheniformis has good application prospect.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various features described in the above embodiments may be combined in any suitable manner without departing from the scope of the invention. The invention is not described in detail in order to avoid unnecessary repetition.
In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.
Claims (13)
1. Bacillus licheniformisBacillus licheniformis) The bacillus licheniformis is characterized in that the preservation number of the bacillus licheniformis is CGMCC NO. 13314.
2. A microbial preparation comprising the Bacillus licheniformis (B) of claim 1Bacillus licheniformis）。
3. The microbial agent according to claim 2, wherein the microbial agent contains a live cell of the Bacillus licheniformis.
4. The microbial inoculum according to claim 2 or 3, wherein the microbial inoculum is a liquid microbial inoculum or a solid microbial inoculum.
5. The microbial inoculum according to claim 4, wherein the microbial inoculum is a solid microbial inoculum.
6. Use of a bacillus licheniformis according to claim 1 and/or a bacterial agent according to any of the claims 2-5 for degrading emetic toxins.
7. The use of claim 6, wherein the vomitoxin is vomitoxin in grain oil and/or feed.
8. A method of degrading emetic toxin, the method comprising: contacting a sample contaminated with a vomitoxin with a bacillus licheniformis according to claim 1 and/or a bacterial agent according to any of the claims 2-5 to degrade the vomitoxin in the sample.
9. The method of claim 8, wherein the vomitoxin-contaminated sample is vomitoxin-contaminated grain oil and/or feed.
10. The method according to claim 8 or 9, wherein the number of the bacillus licheniformis and/or the microbial inoculum is at least 10 per 1g of the sample5CFU。
11. The method of claim 8 or 9, wherein the conditions of the contacting comprise: the temperature is 25-55 deg.C, and pH is 5-11.
12. The method of claim 11, wherein the conditions of the contacting comprise: the temperature is 30-40 deg.C, and pH is 7-10.
13. A kit comprising the Bacillus licheniformis of claim 1 and/or the microbial inoculum of any of claims 2-5.
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| CN109136143A (en) * | 2018-09-11 | 2019-01-04 | 河南工业大学 | One plant of vomitoxin degradation bacteria and its application |
| CN109588539A (en) * | 2018-10-10 | 2019-04-09 | 中山大学 | A kind of reduction forage plant source property raw material is to the dysgenic environment-protecting feed formula of Penaeus Vannmei |
| CN111328962B (en) * | 2018-12-19 | 2022-10-21 | 吉林中粮生化有限公司 | Method for degrading vomitoxin in corn deep processing process |
| CN109744367A (en) * | 2019-03-22 | 2019-05-14 | 东莞市澹一生物科技有限公司 | The method and feedstuff of resource utilization wheat vinasse production feedstuff |
| CN109744368A (en) * | 2019-03-22 | 2019-05-14 | 东莞市澹一生物科技有限公司 | Utilize the method and feedstuff of wheat vinasse production feedstuff |
| CN113755410B (en) * | 2021-11-10 | 2022-03-29 | 中国科学院天津工业生物技术研究所 | Bacillus licheniformis for degrading vomitoxin and application thereof |
| CN115428887A (en) * | 2022-10-26 | 2022-12-06 | 中科检测技术服务(嘉兴)有限公司 | Biological degradation method for mycotoxin in grains or byproducts thereof |
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