CN108251323A - One bacillus licheniformis, the microbial inoculum containing the bacterium and its application, the method and kit of vomitoxin of degrading - Google Patents
One bacillus licheniformis, the microbial inoculum containing the bacterium and its application, the method and kit of vomitoxin of degrading Download PDFInfo
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Abstract
The present invention relates to microorganism field, disclose a bacillus licheniformis, the microbial inoculum containing the bacterium and its application, vomitoxin of degrading method and kit.Specifically, the present invention provides a bacillus licheniformis (Bacillus licheniformis), and the deposit number of the bacillus licheniformis is CGMCC NO.13314.Bacillus licheniformis provided by the invention can efficiently and fast degrade the vomitoxin in grain and oil and/or feed, particularly, even if the vomitoxin content in grain and oil and/feed is up to more than 50ppm, the bacillus licheniformis can also efficiently and fast degrade vomitoxin, and on the palatability of grain and oil and/or feed without influence；Meanwhile it is generated using bacillus licheniformis degradation vomitoxin without toxic products, safety and environmental protection.Therefore, which has a good application prospect.
Description
Technical field
The present invention relates to microorganism fields, and in particular, to a bacillus licheniformis, the microbial inoculum containing the bacterium and its should
With, degradation vomitoxin method and kit.
Background technology
Vomitoxin (vomitoxin), also known as deoxynivalenol (deoxynivalenol, DON), chemical name
Referred to as 3 α, 7 α, 15- trihydroxy grass Fusariumsp -9- alkene -8- ketone, gain the name since it can cause the vomiting of pig, mainly by standing grain
Paddy sickle-like bacteria (Fusarium graminearum), fusarium culmorum (Fusarium culmorum) infection wheat, barley, swallow
The Trichothecenes toxin that the cereal such as wheat, corn generate.In worldwide, vomitoxin is grain, feed, food
One of main contaminated mold toxin seriously affects the health of people and livestock.After people and animals have taken in the food polluted by vomitoxin,
It can lead to the acute poisonings symptom such as apocleisis, vomiting, diarrhea, fever, astasia, slow in reacting, hemopoietic system is damaged when serious
It causes death, serious harm property has caused the most attention of various countries.
Vomitoxin is fairly common to the pollution of China's cereals raw material, and recall rate and detected level are all in mycotoxin
Highest one kind is investigated according to Zhen Yangguang et al. and is shown, the exceeded ratio of Chinese feed and raw material vomitoxin is close to 70%, corn
The exceeding standard rate of middle vomitoxin is 57.1%, and toxin average content is 1.01mg/kg, and highest content is 2.13mg/kg.Due to vomitting
Universal existence, high-content characteristic and acute toxicity and chronic toxicity of the toxin in cereal, feed are spat, reducing or remove its toxicity seems
It is particularly important with it is urgent.At present, domestic and international vomitoxin poison-removing method mainly has Physical, chemical treatment and biological method three greatly
Class.Although physics, chemical method detoxification have been achieved with a degree of success, but still there are detoxification efficiency is limited, Ke Nengzao
Loss into important nutrient, the shortcomings of cost is higher, so as to limit the application of both methods in actual production.
Bioanalysis is mainly the process that toxin degradation is carried out using microorganism or its catabolite, and having can be mild
Under the conditions of reduce the virulence of toxin, the advantages that smaller is influenced on the sensory properties, palatability, nutriment of raw material, meanwhile, also
Have the characteristics that safe and environment-friendly, efficient, it is considered to be best poison-removing method.Therefore, grain and oil are removed using modern biotechnology
Or the research of vomitoxin has a good application prospect in/feed.A plant height is disclosed in patent application CN103243047A
The bacillus subtilis of effect degradation vomitoxin and its application, by 900 μ L bacillus subtilis ANSB471 zymotic fluids and 100 μ L
Vomitoxin (100 μ g/ml) reacts, and 2 hours degradation rates to vomitoxin of reaction are 25%, reacts 24 hours to vomitoxin
Degradation rate for 56%, 48 hours degradation rates to vomitoxin of reaction are 80%, and degradation rate is still to be improved.
In addition, the bioanalysis of existing degradation vomitoxin is in mild condition (such as 25-37 DEG C of temperature and highest mostly
No more than 40 DEG C, pH is 7 or so) under carry out, however, under higher temperature load (as during being transported in a reservoir or
In the case of feed pelleting period) or harsh acid-base property under the conditions of there is no preferable solution, which limits biologies
Application range of the method in vomitoxin of degrading.
Therefore, it is necessary to find it is a kind of efficient and safe, and higher temperature load or harshness acid
The method of biodegradable vomitoxin that can be used under the conditions of alkali.
Invention content
The purpose of the invention is to overcome drawbacks described above present in existing vomitoxin degradation process, one is provided
Bacillus licheniformis, the microbial inoculum containing the bacterium and its application, the method and kit of vomitoxin of degrading.
To achieve these goals, in a first aspect, the present invention provides a bacillus licheniformis (Bacillus
Licheniformis), wherein, the deposit number of the bacillus licheniformis is CGMCC NO.13314.
Second aspect, the present invention also provides a kind of microbial inoculum, wherein, which contains above-mentioned bacillus licheniformis
(Bacillus licheniformis)。
The third aspect, the present invention also provides above-mentioned bacillus licheniformis and/or microbial inoculum answering in vomitoxin of degrading
With.
Fourth aspect, the present invention also provides a kind of method for vomitoxin of degrading, wherein, this method includes：It will be above-mentioned
Bacillus licheniformis and/or above-mentioned microbial inoculum are contacted with the sample that vomitoxin pollutes, with the vomitoxin in sample of degrading.
5th aspect, the present invention provides a kind of kit, the kit includes bacillus subtilis of the present invention
Bacterium and/or microbial inoculum of the present invention.
Bacillus licheniformis provided by the invention can efficiently and fast degrade the vomiting poison in grain and oil and/or feed
Element, particularly, even if the vomitoxin content in grain and oil and/feed is up to more than 50ppm (more preferably at least 100ppm, into one
Step is more preferably at least 150ppm, at least still more preferably 200ppm) when, which can also be efficiently and quick
Ground degradation vomitoxin, and on the palatability of grain and oil and/or feed without influence (i.e. the basic indifference of taste before and after the processing)；Together
When, which is added in the grain and oil and/or feed polluted by vomitoxin, no toxic products generation has
Safety.Therefore, which has a good application prospect.
In addition, bacillus licheniformis provided by the invention also has high temperature and ph stability, particularly in alkaline condition
Under (such as pH be 8-10) it is still with good stability, this further expands the range of its application.
Other features and advantages of the present invention will be described in detail in subsequent specific embodiment part.
Biological deposits
The bacillus licheniformis (Bacillus licheniformis) of the present invention, was deposited on November 16th, 2016
China Committee for Culture Collection of Microorganisms's common micro-organisms center (address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3,
Institute of Microorganism, Academia Sinica, postcode：100101) (depositary institution is abbreviated as CGMCC), deposit number is
CGMCC NO.13314。
Specific embodiment
The specific embodiment of the present invention is described in detail below.It is it should be understood that described herein specific
Embodiment is merely to illustrate and explain the present invention, and is not intended to restrict the invention.
The endpoint of disclosed range and any value are not limited to the accurate range or value herein, these ranges or
Value should be understood to comprising the value close to these ranges or value.For numberical range, between the endpoint value of each range, respectively
It between the endpoint value of a range and individual point value and can be individually combined with each other between point value and obtain one or more
New numberical range, these numberical ranges should be considered as specific open herein.
In a first aspect, the present invention provides a bacillus licheniformis (Bacillus licheniformis), wherein, institute
The deposit number for stating bacillus licheniformis is CGMCC NO.13314.
Bacillus licheniformis provided by the invention is isolated from by vomitoxin contaminated soil sample (being locality Beijing).
The conventional method for new strains separation in this field may be used in the separation of the bacillus licheniformis, such as can be liquid phase
Concentration method or soil ring current method.
Liquid phase concentration method can specifically include：It weighs and is added in right amount by vomitoxin contaminated soil sample equipped with LB liquid
In the triangular flask of culture medium, appropriate bead is added in.Triangular flask is vibrated under 28-37 DEG C, 160-180rpm；Soil is mixed
Liquid is transferred to centrifuge tube, takes source of the supernatant after centrifugation as vomitoxin passivation microorganism.The supernatant is inoculated into
In LB fluid nutrient mediums containing vomitoxin, the shaken cultivation under 28-37 DEG C, 160-180rpm.1- is drawn with aseptic straw
2mL is moved into another enrichment culture triangular flask.So after transfer three times, supernatant is diluted 10 respectively-1、10-2、10-3、
10-4、10-5、10-6、10-7Or 10-8Times, draw the LB for being coated on the vomitoxin containing 50,100,200 or 300mg/L in right amount
Plate streaking is carried out on solid medium, after 28-37 DEG C is cultivated 3-4 days, detaches single bacterium colony.
Soil ring current method can specifically include：It is about 3mm's that 100g, which is weighed, by vomitoxin contaminated soil and appropriate grain size
Sand grains is uniformly mixed, and is placed in the upper strata of circulating device.Lower floor is packed into LB culture mediums 200mL as circulation liquid.Start air compression
Machine starts domestication process, during which periodically adds circulation liquid according to the evaporation situation of circulation liquid.After domestication, by lower floor's circulation
Liquid dilutes 10 respectively-1、10-2、10-3、10-4、10-5、10-6、10-7Or 10-8Times, absorption is coated on containing 50,100,200 in right amount
Or plate streaking is carried out on the LB solid mediums of the vomitoxin of 300mg/L, after 28-37 DEG C is cultivated 3-4 days, detach single bacterium
It falls.
The present inventor has chosen one plant most strong to vomitoxin degradation of bacterium from the bacterial strain filtered out
It has carried out DNA extractions and identification, qualification result is shown, 16srDNA sequences and the bacillus licheniformis (Bacillus of the bacterial strain
Licheniformis) there is 100% homology, it may be determined that the bacterial strain is bacillus licheniformis (Bacillus
Licheniformis), and on November 16th, 2016 it is deposited in China Committee for Culture Collection of Microorganisms's commonly micro- life
Object center, deposit number are CGMCC NO.13314.
Bacillus licheniformis provided by the invention can generate the viable bacteria body of a large amount of bacillus licheniformis by culture.This
Invention is not particularly limited cultural method, as long as the bacillus licheniformis can be made largely to be proliferated by the cultural method
, such as can be according to 105The viable bacteria body of bacillus licheniformis is inoculated in culture medium by the inoculum concentration of CFU/mL, and
Under aerobic condition, after cultivating 12-48 hours at a temperature of 28-38 DEG C, culture solution is obtained.Wherein, the culture medium can be
Culture medium commonly used in the art, for example, can be LB fluid nutrient mediums (0.8-1 weight % peptones, 0.5-0.8 weights
Measure % dusty yeasts, 1-1.5 weight % sodium chloride；PH is 6.8-7.0；Cultivation temperature is 28-38 DEG C) or nutrient broth medium
(0.8-1 weight % peptones, 0.3-0.5 weight % beef extracts, 0.5-0.8 weight % sodium chloride；PH is 7.2-7.6；Culture temperature
Spend is 28-38 DEG C), preferably LB fluid nutrient mediums.
The present invention can further detach the thalline of the bacillus licheniformis in above-mentioned culture solution, to the method for the separation
Be not particularly limited, if thalline can be enriched with from culture solution, such as can be by centrifuging and/or filtering method
It realizes, the condition of the centrifugation and the filtering can be the normal condition of this field, this is well known to those skilled in the art,
Details are not described herein.
Second aspect, the present invention provides a kind of microbial inoculum, wherein, which contains above-mentioned bacillus licheniformis
(Bacillus licheniformis)。
In the present invention, the concentration of bacillus licheniformis in the microbial inoculum is not particularly limited, it can be according to specific
Situation specifically selected.
According to the present invention, the microbial inoculum contains the viable bacteria body of the bacillus licheniformis and/or dead thalline.Preferably, institute
State viable bacteria body or the mixing thalline of viable bacteria body and dead thalline that microbial inoculum contains the bacillus licheniformis.When the microbial inoculum contains
During the mixing thalline of the viable bacteria body of the bacillus licheniformis and dead thalline, preferably the quantity of viable bacteria body is higher than the number of dead thalline
Amount.Most preferably described microbial inoculum contains the viable bacteria body of the bacillus licheniformis.
According to the present invention, the dosage form of the microbial inoculum is not particularly limited, it can be according to the difference of intended purpose, by it
Different dosage forms is prepared into, and adds the ingredients such as corresponding excipient, such as the microbial inoculum can be that liquid microbial inoculum (such as can be with
For bacterium solution) and/or solid-state microbial inoculum (such as can be thalline) after freeze-drying, preferably solid-state microbial inoculum.Wherein, in which kind of dosage form
Add which kind of excipient is well known to those skilled in the art in microbial inoculum, in this not go into detail.
In addition, the present inventor has found in the course of the research, although bacillus licheniformis provided by the invention be
Screening obtains under conditions of high concentration vomitoxin (at least 50ppm), but it is dirty to other mycotoxins common in soil
Dye (such as vomitoxin, aflatoxin, fumonisin, ochratoxin, T2 toxin) has certain degradation.
The third aspect, the present invention also provides above-mentioned bacillus licheniformis and/or above-mentioned microbial inoculum in vomitoxin of degrading
Application, preferably degradation grain and oil and/or feed in vomitoxin in application.
Fourth aspect, the present invention also provides a kind of method for vomitoxin of degrading, wherein, this method includes：It will be above-mentioned
Bacillus licheniformis and/or above-mentioned microbial inoculum are contacted with the sample that vomitoxin pollutes, with the vomitoxin in sample of degrading.
According to the present invention, the sample of the vomitoxin pollution can be the grain and oil and/or feed of vomitoxin pollution.
In situations where it is preferred, on the basis of the total weight of the sample of vomitoxin pollution, the vomitoxin
Content is at least 1ppm, further preferably at least 50ppm, still more preferably at least 100ppm, still further preferably extremely
It is less 150ppm, then is still more preferably at least 200ppm.In the present invention, when the pending sample is liquid,
That " ppm " is represented is " μ g/mL "；When the pending sample is solid, that " ppm " is represented is " μ g/g ".
In the present invention, term " grain and oil " refers to the grains such as cereal, beans and oil plant and its processing finished product and semi-finished product
General designation, particularly relate to the edible product of the mankind.For example, the grain and oil can be that the common mankind in this field are edible
Grain oil product, specifically, the grain and oil can include cereal and its agricultural and sideline product, oil ＆ fat product, drinks, milk and its system
At least one of product etc..
In the present invention, term " feed " refers to the general name of the food of agricultural or animal husbandry domesticated animal.For example, the feeding
Material can be food used in the common raising animal in this field, and specifically, the feed can include：A) cereal, such as
Granule cereal (such as wheat, barley, naked barley, oat and combination thereof) and/or big grain cereal such as maize or sorghum；B) come
From the by-product of cereal, such as corn protein powder, distiller's dried grain and soluble matter (DDGS), wheat bran, sizing, wheat wheat-middlings, rice
Chaff, rice husk, oat shell, palm kernel and citrus pulp；C) ensilage；D) protein derived from following source：Such as soybean, Xiang
Certain herbaceous plants with big flowers, peanut, lupin, pea, broad bean, cotton, Canola, fish meal, dry plasma albumen, meat and bone meal, potato protein, breast
Clearly, copra, sesame；E) oil ＆ fat obtained from plant and animal source；F) minerals and vitamins.
In the present invention, the grain and oil or feed can also contain physiologically acceptable carrier, wherein, the physiology
Upper acceptable carrier is selected from least one of following substance：Maltodextrin, lime stone (calcium carbonate), cyclodextrin, wheat,
Wheat bran or wheat component, rice or rice bran, sucrose, starch, Na2SO4With talcum powder and their mixture.
In the present invention, the dosage form of the bacillus licheniformis and/or microbial inoculum is not particularly limited, it can be according to pre-
Determine the difference of purposes, be prepared into different dosage forms, and add the ingredients such as corresponding excipient, such as the lichens gemma bar
Bacterium and/or microbial inoculum can be liquid and/or solid-state.Wherein, which kind of excipient is added in which kind of dosage form as people in the art
Well known to member, in this not go into detail.
According to the present invention, relative to sample described in 1g, the thalline number in the bacillus licheniformis and/or the microbial inoculum is extremely
It is 10 less5CFU, it is preferable that the viable bacteria body number in the bacillus licheniformis and/or the microbial inoculum is at least 105CFU.In this hair
In bright, thalline number can be measured according to GB4789.2-94.
According to the present invention, the condition of the contact can include：Temperature is 25-55 DEG C, pH value 5-11, time 1-
48h；Preferably, temperature is 30-40 DEG C, pH value 7-10, time 12-36h.
In the present invention, when the sample is solid, the pH value is surveyed according to the method for GB/T 12456-2008
It is fixed.
5th aspect, the present invention provides a kind of kit, the kit includes bacillus subtilis of the present invention
Bacterium and/or microbial inoculum of the present invention.
Embodiment
The present invention will be described in detail by way of examples below.
In following preparation example, preparation comparative example, embodiment and comparative example：
Vomitoxin standard items are purchased from Sigma companies, and the experiment material used in remaining is from often unless otherwise specified
Rule biochemical reagents shop is commercially available.
LB fluid nutrient mediums：10g peptones, 5g dusty yeasts, 10g sodium chloride, deionized water are settled to 1L, adjust pH to 7.
LB solid mediums：10g peptones, 5g dusty yeasts, 10g sodium chloride, 16g agar powders, deionized water are settled to 1L,
Adjust pH to 7.
Bacterial strain provided by the invention is bacillus licheniformis (Bacillus licheniformis), and in November, 2016
It is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (address within 16th：The Chaoyang District, Beijing City North Star
The institute 3 of West Road 1, Institute of Microorganism, Academia Sinica, postcode：100101) (depositary institution is abbreviated as CGMCC),
Deposit number is CGMCC NO.13314.
Reference strains 1 are the bacillus licheniformis (Bacillus that deposit number is CGMCC NO.1.807
Licheniformis), purchased from CGMCC.
Reference strains 2 are the bacillus subtilis that the deposit number disclosed in CN103243047A is CGMCC NO.7344
(Bacillus subtilis), purchased from CGMCC.
Thalline number is measured according to GB4789.2-94.
According to《Measure affine in immunity column purification-height of deoxynivalenol in GB/T 30956-2014 feeds
Effect liquid phase chromatogram method》Measure vomitoxin content.
The degradation rate (%) of vomitoxin=(vomit poison before reaction in sample after quality-reaction of vomitoxin in sample
The quality of element) before/reaction in sample vomitoxin quality × 100%.
Preparation example 1
After the bacillus licheniformis that deposit number provided by the invention is CGMCC NO.13314 is activated, with 1 volume %
Inoculum concentration be inoculated in through 121 DEG C sterilizing 15min LB fluid nutrient mediums in, the shaken cultivation under the conditions of 160rpm, 37 DEG C, training
The foster time so that cell concentration is (1 ± 0.1) × 10 in bacterium solution5CFU/mL.While dry powder is made, contained by every gram of dry powder
Total viable count is (1 ± 0.1) × 105CFU。
Prepare comparative example 1
According to the method for preparation example 1, the difference is that, with the bacillus licheniformis that deposit number is CGMCC NO.1.807
(Bacillus licheniformis) replaces the lichens gemma bar that deposit number provided by the invention is CGMCC NO.13314
Bacterium.
Prepare comparative example 2
According to the method for preparation example 1, the difference is that, it is CGMCC with the deposit number disclosed in CN103243047A
It is CGMCC that the bacillus subtilis (Bacillus subtilis) of NO.7344, which replaces deposit number provided by the invention,
The bacillus licheniformis of NO.13314.
Embodiment 1
The present embodiment is used for the ability for illustrating bacillus licheniformis degradation vomitoxin provided by the invention.
1. degradation time measures
The bacterium solution that the culture of 900 μ L preparation examples 1 obtains is taken to be placed in 1.5mL centrifuge tubes, adds in vomitoxin standard solution,
It is uniformly mixed, obtains mixed liquor, wherein, the final concentration of 50ppm of vomitoxin in mixed liquor.
By above-mentioned mixed liquor in 37 DEG C, pH to react under conditions of 7, respectively in reaction 1h, 12h, for 24 hours, 36h, 48h
When negate the residual that the 20 μ L of sample that answer are used to detect vomitoxin.The results are shown in Table 1.
By the result of table 1 it is found that reaction can make the degradation rate of vomitoxin up to more than 90% for 24 hours.Therefore, in the case where connecing
Using for 24 hours as the reaction time in the experiment come.
2. thermal stability determination
Obtained bacterium solution will be cultivated respectively at 40 DEG C, 45 DEG C, 50 DEG C and 55 DEG C, under conditions of pH is 7 after processing 30min,
The 900 treated bacterium solutions of μ L is taken to be placed in 1.5mL centrifuge tubes respectively, add in vomitoxin standard solution, are uniformly mixed,
Mixed liquor is obtained, wherein, the final concentration of 50ppm of vomitoxin in mixed liquor.Then it is in 37 DEG C, pH by above-mentioned mixed liquor again
It is reacted under conditions of 7 for 24 hours, after the completion of reaction, takes 20 μ L of sample for detecting the residual of vomitoxin.The results are shown in Table 2.
It is found that bacterium solution preserves 30min at 40-55 DEG C still there is higher vomitoxin degradation to live by the result of table 2
Property.
3. ph stability measures
It is 5,6,7,8,9,10 and 11 that obtained bacterium solution, which will be cultivated, respectively at pH, after handling 30min at 37 DEG C, is taken respectively
The 900 treated bacterium solutions of μ L are placed in 1.5mL centrifuge tubes, add in vomitoxin standard solution, are uniformly mixed, are mixed
Liquid is closed, wherein, the final concentration of 50ppm of vomitoxin in mixed liquor.Then again by above-mentioned mixed liquor in 37 DEG C, the item that pH is 7
It is reacted under part for 24 hours, after the completion of reaction, takes 20 μ L of sample for detecting the residual of vomitoxin.The results are shown in Table 3.
Still there is higher vomitoxin to degrade it is found that bacterium solution preserves 30min in the case where pH value is 5-11 by the result of table 3
Activity.
Comparative example 1
Method according to embodiment 1 is measured, the difference is that bacterium solution is prepared using preparation comparative example 1 respectively
Instead of bacterium solution used in embodiment 1.
The results are shown in Table 1 for degradation time measure, and the results are shown in Table 2 for thermal stability determination, ph stability
The results are shown in Table 3.
Comparative example 2
Method according to embodiment 1 is measured, the difference is that bacterium solution is prepared using preparation comparative example 2 respectively
Instead of bacterium solution used in embodiment 1.
The results are shown in Table 1 for degradation time measure, and the results are shown in Table 2 for thermal stability determination, ph stability
The results are shown in Table 3.
Table 1
Table 2
Table 3
Testing example 1
The dry powder 10g obtained by preparation example 1 with the corn flour that 1kg pollutions there are 200ppm vomitoxins to pollute is mixed, is added
Enter 1kg distilled water, do three repetitions, it is detoxification under conditions of 71 day to be uniformly mixed after 37 DEG C, pH value.Then according to《GB/T
Measure affine in immunity column purification-high performance liquid chromatography of deoxynivalenol in 30956-2014 feeds》At measure
The content of reason vomitoxin in each sample after 1 day, and calculate degradation rate.It is computed, the degradation rate of vomitoxin is in corn flour
90.8%.Also, the addition of dry powder does not generate the palatability of corn flour any influence.
Feed pig 14 days using corn flour after processing above, the results showed that, during this period, the diet of pig, drinking-water and
Daily routines are acted normally, and the weight of pig is also always maintained at the trend of steady growth.It these results suggest that preparation example 1
Obtained dry powder is added in corn flour for swine rearing, has safety.
Testing example 2
According to the method for testing example 1, the difference is that, the content of vomitoxin is 100ppm in corn flour, after testing
The degradation rate of vomitoxin is 95.9%.Also, the addition of dry powder does not generate the palatability of corn flour any influence.
Feed pig 14 days using corn flour after processing above, the results showed that, during this period, the diet of pig, drinking-water and
Daily routines are acted normally, and the weight of pig is also always maintained at the trend of steady growth.It these results suggest that and add dry powder
It adds in corn flour for swine rearing, there is safety.
Testing example 3
According to the method for testing example 1, the difference is that, the content of vomitoxin is 50ppm in corn flour, is vomitted after testing
The degradation rate for spitting toxin is 100%.Also, the addition of dry powder does not generate the palatability of corn flour any influence.
Feed pig 14 days using corn flour after processing above, the results showed that, during this period, the diet of pig, drinking-water and
Daily routines are acted normally, and the weight of pig is also always maintained at the trend of steady growth.It these results suggest that and add dry powder
It adds in corn flour for swine rearing, there is safety.
Test comparison example 1
It is carried out according to the method for testing example 1, the difference is that being replaced using the dry powder that preparation comparative example 1 obtains real
Apply the dry powder used in example 1.The degradation rate of vomitoxin is 16.8% after testing.
Test comparison example 2
It is carried out according to the method for testing example 1, the difference is that being replaced using the dry powder that preparation comparative example 2 obtains real
Apply the dry powder used in example 1.The degradation rate of vomitoxin is 18.1% after testing.
By above example 1 it is found that bacillus licheniformis provided by the invention can compared with the result of comparative example 1-2
With the vomitoxin in efficiently and fast degrade grain and oil and/or feed, and the bacillus licheniformis also has high temperature and acid
Alkaline stability, (such as pH is 8-10) is still with good stability particularly under alkaline condition.
By more than testing example 1 compared with the result of test comparison example 1-2 it is found that even if by provided by the invention
Bacillus licheniformis is applied to vomitoxin content and is up in the grain and oil and/feed of more than 50ppm, which also may be used
With vomitoxin of efficiently and fast degrading, and on the palatabilities of grain and oil and/or feed without influencing (i.e. taste base before and after the processing
This indifference)；Meanwhile the bacillus licheniformis is added in the grain and oil and/or feed polluted by vomitoxin, without toxic production
Object generates, and has safety.Therefore, which has a good application prospect.
The preferred embodiment of the present invention has been described above in detail, still, during present invention is not limited to the embodiments described above
Detail, within the scope of the technical concept of the present invention, a variety of simple variants can be carried out to technical scheme of the present invention, this
A little simple variants all belong to the scope of protection of the present invention.
It is further to note that specific technical features described in the above specific embodiments, in not lance
In the case of shield, it can be combined by any suitable means.In order to avoid unnecessary repetition, the present invention to it is various can
The combination of energy no longer separately illustrates.
In addition, various embodiments of the present invention can be combined randomly, as long as it is without prejudice to originally
The thought of invention, it should also be regarded as the disclosure of the present invention.
Claims (10)
- A 1. bacillus licheniformis (Bacillus licheniformis), which is characterized in that the bacillus licheniformis Deposit number is CGMCC NO.13314.
- 2. a kind of microbial inoculum, which is characterized in that the microbial inoculum contains bacillus licheniformis (Bacillus described in claim 1 licheniformis)。
- 3. microbial inoculum according to claim 2, wherein, the microbial inoculum contain the bacillus licheniformis viable bacteria body and/or Dead thalline, preferably viable bacteria body.
- 4. the microbial inoculum according to Claims 2 or 3, wherein, the microbial inoculum is liquid microbial inoculum and/or solid-state microbial inoculum, preferably Solid-state microbial inoculum.
- 5. the microbial inoculum in bacillus licheniformis described in claim 1 and/or claim 2-4 described in any one is vomitted in degradation Spit the application in toxin, the preferably application in the vomitoxin in degradation grain and oil and/or feed.
- A kind of 6. method for vomitoxin of degrading, which is characterized in that this method includes：By lichens gemma described in claim 1 Microbial inoculum in bacillus and/or claim 2-4 described in any one is contacted with the sample that vomitoxin pollutes, in sample of degrading Vomitoxin.
- 7. according to the method described in claim 6, wherein, the sample of the vomitoxin pollution is the grain and oil of vomitoxin pollution And/or feed.
- 8. the method described according to claim 6 or 7, wherein, relative to sample described in 1g, the bacillus licheniformis and/or Thalline number in the microbial inoculum is at least 105CFU。
- 9. according to the method described in any one in claim 6-8, wherein, the condition of the contact includes：Temperature is 25-55 DEG C, preferably 30-40 DEG C；PH value is 5-11, preferably 7-10.
- 10. a kind of kit, which is characterized in that including bacillus licheniformis described in claim 1 and/or claim 2-4 Microbial inoculum described in middle any one.
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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 |
| CN111328962A (en) * | 2018-12-19 | 2020-06-26 | 吉林中粮生化有限公司 | Method for degrading vomitoxin in corn deep processing process |
| 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 |
| CN113755410A (en) * | 2021-11-10 | 2021-12-07 | 中国科学院天津工业生物技术研究所 | Bacillus licheniformis for degrading vomitoxin and application thereof |
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| 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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