CN103355356A - Use of Bacillus subtilis fermentation liquid to inhibit the growth of Penicillium and Aspergillus niger - Google Patents
Use of Bacillus subtilis fermentation liquid to inhibit the growth of Penicillium and Aspergillus niger Download PDFInfo
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- CN103355356A CN103355356A CN2013100308944A CN201310030894A CN103355356A CN 103355356 A CN103355356 A CN 103355356A CN 2013100308944 A CN2013100308944 A CN 2013100308944A CN 201310030894 A CN201310030894 A CN 201310030894A CN 103355356 A CN103355356 A CN 103355356A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 16
- 230000004151 fermentation Effects 0.000 title claims abstract description 16
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- 239000012530 fluid Substances 0.000 claims description 13
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Abstract
本发明公开一种枯草芽孢杆菌(Bacillussubtilis)菌株(CICCNo.10321)发酵液抑制青霉和黑曲霉生长的用途，枯草芽孢杆菌(Bacillussubtilis)菌株(CICCNo.10321)对青霉和黑曲霉具有抑制作用（抑制青霉和黑曲霉的抑菌圈大于20cm以上），且抑制效果好，持续时间长。该菌株培养简单，周期短，成本低，适合于作为霉菌抑制剂，抑制饲料等易霉变物品中青霉和黑曲霉的生长，有效降低霉菌毒素在动物性食品中的残留。
The invention discloses a use of a fermentation liquid of a Bacillus subtilis strain (CICC No. 10321) for inhibiting the growth of Penicillium and Aspergillus niger. The Bacillus subtilis strain (CICC No. 10321) has an inhibitory effect on Penicillium and Aspergillus niger (the inhibition zone of Penicillium and Aspergillus niger is greater than 20 cm), and the inhibitory effect is good and the duration is long. The strain is simple to culture, has a short cycle, and is low in cost. It is suitable as a mold inhibitor to inhibit the growth of Penicillium and Aspergillus niger in feed and other items that are prone to mold, and effectively reduce the residue of mycotoxins in animal food.
Description
Technical field
The present invention relates to a kind of new purposes of fermentation of bacillus subtilis liquid, especially a kind of fermentation of bacillus subtilis liquid suppresses the purposes of mould and Aspergillus Niger Growth.
Background technology
Mycotoxin is the poisonous secondary metabolite that mould produces in process of growth, mainly comprises aflatoxin, Trichothecenes toxin (such as T-2 toxin, neo-solaniol and vomitoxin), zearalenone, ochratoxin A and FT etc.According to the statistics made by the departments concerned, in existing 194 parts of feeds and raw material corn, the wheat bran, the pollution rate of vomitoxin, T-2 toxin and zearalenone is respectively 50.0%, 18.1% and the biodegradable progress of 26%(Mycotoxins in Feed [J]. Scientia Agricultura Sinica 2012,45(1): 153-158).Mycotoxin mainly produces toxicity to animal's liver and kidney, behind the feed that animal ingestion goes mouldy, can cause a series of body reactions, be called " background color is sick ", main manifestations is that the animal body feed intake reduces poor growth, sexual maturity is postponed, and sexual desire descends, sperm concentration and Quality Down, litter size reduces, the lactation ability reduces, miscarriage, and immunity function descends, the incidence of disease improves, the illnesss such as bloody stool.At present, be widely used the mineral matter such as kaolin both at home and abroad as taking off mould dose, but this kind takes off the mould dose of mycotoxin in can only absorbent articles, can not suppress fungus growth in the feed preparation process, and in absorbing mycotoxin the also trace element such as vitamin, mineral matter and some feed addictives etc. in the adsorption feed, significantly reduced the nutritive value of feed and the conversion ratio of feed in animal body etc.In recent years, utilize the research of the tunning mould fungus inhibition growth of microorganism to receive many scholars' concern, domestic research mainly contain Li Junxia (screening of Aflatoxin B 1 Degradation Strains and evaluation [J]. Scientia Agricultura Sinica, 2008,41(5): 1459-1463) etc., the report of external mould fungus inhibition mainly concentrates on Bifidobacterium, saccharomycete etc., and focuses mostly in the research of aspergillus flavus.But, also do not have up to now the relevant report about fermentation of bacillus subtilis liquid inhibition mould and Aspergillus Niger Growth.
Summary of the invention
The present invention be found bacillus subtilis (
Bacillus subtilis) (CICC No.10321) have and suppress mould and aspergillus niger effect, thereby invented the purposes that fermentation of bacillus subtilis liquid suppresses mould and Aspergillus Niger Growth.
Technical solution of the present invention is: a kind of fermentation of bacillus subtilis liquid suppresses the purposes of mould and Aspergillus Niger Growth, described bacillus subtilis be bacillus subtilis (
Bacillus subtilis) bacterial strain (CICCNo.10321).
Described fermentation of bacillus subtilis liquid is to ferment as follows:
A. the scraping bacillus subtilis (
Bacillus subtilis) the rear agar plate line of bacterial strain (CICCNo.10321), cultivating after 24 hours the single colony inoculation of picking cultivated in liquid nutrient medium 24 hours, described liquid nutrient medium is to contain in every premium on currency: tryptone 18 ~ 20g, dusty yeast 8 ~ 10g, glucose 6 ~ 8, dipotassium hydrogen phosphate 4 ~ 6g, pH value are 6.5 ~ 7.5;
B. bacterium liquid is inoculated in again in the liquid nutrient medium identical with a step, inoculum concentration 5 ~ 10%, 37 ℃, 180 ~ 220r/min cultivates 24 ~ 48h;
C. with gained culture fluid 8000r/min, 5min is centrifugal, and 0.22 μ m membrane filtration gets supernatant, i.e. zymotic fluid.
The present invention found bacillus subtilis (
Bacillus subtilis) bacterial strain (CICCNo.10321) is to mould and aspergillus niger inhibited (suppressing the inhibition zone of mould and aspergillus niger greater than more than the 20cm), and inhibition is good, longer duration.This strain culturing is simple, and the cycle is short, and cost is low, is suitable for as mould inhibitor, suppresses the growth of mould and aspergillus niger in the article that easily go mouldy such as feed, effectively reduces residual in animal food of mycotoxin.
Description of drawings
Fig. 1 is that embodiment of the invention fermentation of bacillus subtilis liquid is to the drug sensitive experiment photo of mould.
Fig. 2 is that embodiment of the invention fermentation of bacillus subtilis liquid is to the drug sensitive experiment photo of aspergillus niger.
Fig. 3 is that embodiment of the invention fermentation of bacillus subtilis liquid and other fermentation of bacillus subtilis liquid are to four kinds of mould inhibition histograms (the different strains zymotic fluid is to the fungistatic effect histogram of four kinds of moulds).
Embodiment
The preparation bacillus subtilis (
Bacillus subtilis) bacterial strain (CICCNo.10321) zymotic fluid:
A. scraping be purchased from Chinese industrial microorganism fungus kind preservation administrative center bacillus subtilis (
Bacillus subtilis) the rear agar plate line of bacterial strain (CICCNo.10321), cultivating after 24 hours the single colony inoculation of picking cultivated in liquid nutrient medium 24 hours, described liquid nutrient medium is to contain in every premium on currency: tryptone 18 ~ 20g, dusty yeast 8 ~ 10g, glucose 6 ~ 8, dipotassium hydrogen phosphate 4 ~ 6g, pH value are 6.5 ~ 7.5;
B. bacterium liquid is inoculated in again in the liquid nutrient medium identical with a step, inoculum concentration 5 ~ 10%, 37 ℃, 180 ~ 220r/min cultivates 24 ~ 48h;
C. with gained culture fluid 8000r/min, 5min is centrifugal, and 0.22 μ m membrane filtration degerming gets supernatant, i.e. zymotic fluid.
Bacillus subtilis (
Bacillus subtilis) bacterial strain (CICCNo.10321) zymotic fluid is to the drug sensitive experiment (punch method) of mould:
Dip an amount of Penicillium notatum suspension with aseptic swab stick, evenly coat the Rose Bengal Sodium flat board, in coated Rose Bengal Sodium plate, punch behind the card punch flame disinfection, abandon or adopt hole middle plateform piece, add 0.2 μ l zymotic fluid behind the agar back cover, measure the inhibition zone size behind 37 ℃ of cultivation 48h.
The result is as shown in Figure 1: among the figure 5,6 be the zymotic fluid of embodiment of the invention CICC No.10321 to the inhibition zone of mould, inhibition zone is transparent, without spore, diameter 23mm.
Bacillus subtilis (
Bacillus subtilis) bacterial strain (CICCNo.10321) zymotic fluid is to the drug sensitive experiment (punch method) of aspergillus niger:
Dip an amount of black-koji mould suspension with aseptic swab stick, evenly coat the Rose Bengal Sodium flat board, in coated Rose Bengal Sodium plate, punch behind the card punch flame disinfection, abandon or adopt hole middle plateform piece, add 0.2 μ l zymotic fluid behind the agar back cover, measure the inhibition zone size behind 37 ℃ of cultivation 72h.
The result is as shown in Figure 2: 5,6 is the inhibition zone of aspergillus niger of the zymotic fluid of embodiment of the invention CICC No.10321 among the figure, and inhibition zone is transparent, without spore, and diameter 21mm.
With same method detect preserving number be respectively CICC No.10002, CICC No.10066, CICC No.10282, CICC No.10339, CICC No.21145 and bacillus subtilis of the present invention (
Bacillus subtilis) zymotic fluid to the inhibition histograms of different moulds (mould, aspergillus niger, Mucor, aspergillus flavus) as shown in Figure 1, the result represents, bacillus subtilis of the present invention (
Bacillus subtilis) bacterial strain (CICCNo.10321) zymotic fluid is remarkable to mould and aspergillus niger inhibition.
Claims (2)
1. a fermentation of bacillus subtilis liquid suppresses the purposes of mould and Aspergillus Niger Growth, it is characterized in that: described bacillus subtilis be bacillus subtilis (
Bacillus subtilis) bacterial strain (CICCNo.10321).
2. fermentation of bacillus subtilis liquid according to claim 1 suppresses the purposes of mould and Aspergillus Niger Growth, it is characterized in that described fermentation of bacillus subtilis liquid is to ferment as follows:
A. the scraping bacillus subtilis (
Bacillus subtilis) the rear agar plate line of bacterial strain (CICCNo.10321), cultivating after 24 hours the single colony inoculation of picking cultivated in liquid nutrient medium 24 hours, described liquid nutrient medium is to contain in every premium on currency: tryptone 18 ~ 20g, dusty yeast 8 ~ 10g, glucose 6 ~ 8, dipotassium hydrogen phosphate 4 ~ 6g, pH value are 6.5 ~ 7.5;
B. bacterium liquid is inoculated in again in the liquid nutrient medium identical with a step, inoculum concentration 5 ~ 10%, 37 ℃, 180 ~ 220r/min cultivates 24 ~ 48h;
C. with gained culture fluid 8000r/min, 5min is centrifugal, and 0.22 μ m membrane filtration gets supernatant, i.e. zymotic fluid.
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| CN103859165A (en) * | 2014-03-06 | 2014-06-18 | 苏州万生源生物科技有限公司 | Mycotoxin purification agent for feed and preparation method and application of agent |
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| CN106721082A (en) * | 2016-12-20 | 2017-05-31 | 福州大学 | A kind of preparation method and applications of bacillus subtilis metabolite |
| CN109486707A (en) * | 2018-11-23 | 2019-03-19 | 北京林业大学 | A kind of bacillus subtilis strain and its application |
| CN109645029A (en) * | 2018-10-12 | 2019-04-19 | 北京师范大学 | Preparation and application of a kind of cultural relic mold and mildew inhibitor |
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| CN110184226A (en) * | 2019-06-28 | 2019-08-30 | 湖南农业大学 | Bacillus Mangrove-2008, microbial inoculum and its purposes and method in inhibition Aspergillus Niger Growth |
| CN110283746A (en) * | 2019-06-28 | 2019-09-27 | 湖南农业大学 | Bacillus Mangrove-2001, microbial inoculum and its purposes and method in inhibition Aspergillus Niger Growth |
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