CN109619289A - Bacillus coagulans ACCC10229 is reducing the application in alfalfa ensilage butyric acid content - Google Patents
Bacillus coagulans ACCC10229 is reducing the application in alfalfa ensilage butyric acid content Download PDFInfo
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Abstract
The present invention provides bacillus coagulans ACCC10229 to reduce the application in alfalfa ensilage butyric acid content, belongs to field of feed.In the present invention, by adding bacillus coagulans ACCC10229 fermentation liquid during alfalfa ensilage, to reduce the content of butyric acid in alfalfa ensilage.Embodiment the result shows that: bacillus coagulans ACCC10229 fermentation liquid can make butyric acid in alfalfa ensilage content reduce by 73.64%~82.26%.
Description
Technical field
The invention belongs to field of feed, and in particular to bacillus coagulans ACCC10229 is reducing alfalfa ensilage fourth
Application in acid content.
Background technique
Alfalfa (Medicago sativa) is widely used high-quality protein class herbage in milk cattle cultivating, is known as
King of Pasture.Often is overlapped with rainy season in the more region clover harvest of Summer Rainfall in China weather category rain heat same season, lose compared with
Greatly.Ensiling is difficult preferred method of harvesting and storing in solution clover rainy season as one of the effective means for promoting feeding value of forage.Lucerne
Mu ensiling (alfalfa silage), soft and succulency, palatability are good and digestibility is high, are convenient for long-term preservation.
For poultry, pig sheep, the butyric acid in feed can be improved nutrient digestibility, increase digestive ferment secretion, adjustment intestines
Road microbiota improves epithelial cell integrality and enhancing system of defense function, to improve growth of animal and productivity
Can, but the butyric acid in feed is unfavorable for the digestion and absorption of ox.The degree of butyric fermentation is identification ox ensilage quality
Important symbol, butyric acid content is higher, and ensilage quality is lower.
Summary of the invention
In view of this, the present invention provides bacillus coagulans ACCC10229 in reducing alfalfa ensilage butyric acid content
Using when preparing alfalfa ensilage, addition bacillus coagulans ACCC10229, can be effectively reduced butyric acid in alfalfa ensilage
Content improves the Forage quality of alfalfa ensilage.
To solve the above-mentioned problems, the present invention provides following technical schemes:
The present invention provides bacillus coagulans ACCC10229 to reduce the application in alfalfa ensilage butyric acid content.
Preferably, after the bacillus coagulans ACCC10229 being fermented, obtained fermentation liquid is added to clover
In prepare alfalfa ensilage.
Preferably, the preparation method of the bacillus coagulans ACCC10229 fermentation liquid includes: by bacillus coagulans
ACCC10229 bacterium solution is inoculated into liquid state fermentation culture medium, and at 30~40 DEG C, ferment 20~28h under conditions of 80~120rpm,
Obtain bacillus coagulans ACCC10229 fermentation liquid；The liquid state fermentation culture medium takes water as a solvent, the group including following concentration
Point: peptone 12g/L, powdered beef 12g/L, yeast powder 6g/L, glucose 24g/L, dipotassium hydrogen phosphate 2g/L, diammonium hydrogen citrate
2g/L, sodium acetate 5g/L, magnesium sulfate 0.58g/L, manganese sulfate 0.25g/L and tween 5mL/L, the pH of the liquid state fermentation culture medium
Value is 5.8~6.0.
Preferably, the viable count of the bacillus coagulans ACCC10229 bacterium solution is 1.0 × 108~1.0 × 1010CFU/
g。
Preferably, the inoculum concentration is the 5%~15% of liquid state fermentation total weight of medium.
Preferably, the preparation method of the alfalfa ensilage, includes the following steps:
1) bacillus coagulans ACCC10229 is fermented, obtains bacillus coagulans ACCC10229 fermentation liquid；
2) clover is mixed with the bacillus coagulans ACCC10229 fermentation liquid of the step 1), obtains total mixture；
3) total mixture of the step 2) is sealed 40~50d of fermentation, obtains alfalfa ensilage.
Preferably, clover and the mass volume ratio of bacillus coagulans ACCC10229 fermentation liquid are in the step 2)
1Kg:1.4~1.6mL.
It preferably, further include being diluted bacillus coagulans ACCC10229 fermentation liquid before being mixed described in step 2)；
The diluted multiple is 3~5 times.
It preferably, further include the clover section that clover is cut into 2~5cm before being mixed described in step 2).
Preferably, the temperature being sealed by fermentation in the step 3) is 20~30 DEG C.
The present invention provides bacillus coagulans ACCC10229 to reduce the application in alfalfa ensilage butyric acid content.At this
In invention, bacillus coagulans ACCC10229 is homofermentative lactic bacteria, and bacillus coagulans are added in ensilage
After ACCC10229 fermentation liquid, homofermentative lactic is carried out, increases fermenting speed, lactic acid content increases, and homolactic bacterium occupies excellent
Gesture, inhibits the growth of clostridium butyricum and enterobacteria, so that butyric acid and acetic acid content be made to decline, butyric acid contains in reduction alfalfa ensilage
Amount.Embodiment is the result shows that addition bacillus coagulans ACCC10229 fermentation liquid can be significant during preparing alfalfa ensilage
The content for reducing butyric acid in alfalfa ensilage, compared to control group 1~4, butyric acid content reduces by 73.64%~82.26%.
Specific embodiment
The present invention provides bacillus coagulans ACCC10229 to reduce the application in alfalfa ensilage butyric acid content.
In the present invention, the bacillus coagulans ACCC10229 is bought from Chinese agriculture Microbiological Culture Collection management
Center.
In the present invention, it is preferred to which obtained fermentation liquid is added after the bacillus coagulans ACCC10229 is fermented
It is added in clover and prepares alfalfa ensilage.
In the present invention, the preparation method of the bacillus coagulans ACCC10229 fermentation liquid preferably includes: will condense bud
Spore bacillus ACCC10229 bacterium solution is inoculated into liquid state fermentation culture medium, at 30~40 DEG C, ferments 20 under conditions of 80~120rpm
~28h obtains bacillus coagulans ACCC10229 fermentation liquid；The liquid state fermentation culture medium takes water as a solvent, including following dense
The component of degree: peptone 12g/L, powdered beef 12g/L, yeast powder 6g/L, glucose 24g/L, dipotassium hydrogen phosphate 2g/L, citric acid
Hydrogen diammonium 2g/L, sodium acetate 5g/L, magnesium sulfate 0.58g/L, manganese sulfate 0.25g/L and tween 5mL/L.In the present invention, described
The pH value of liquid state fermentation culture medium is preferably 5.8~6.0, and more preferably 5.9.The present invention is to each in the liquid state fermentation culture medium
The source of component is not particularly limited, using this field conventional commercial product.
In the present invention, the amount of the inoculation of the bacillus coagulans ACCC10229 bacterium solution is preferably liquid state fermentation culture
The 5%~15% of base total weight, more preferably 10%.In the present invention, the work of the bacillus coagulans ACCC10229 bacterium solution
Bacterium number is preferably 1.0 × 108~× 1.0 × 1010CFU/g, more preferably 1.0 × 109CFU/g.In the present invention, the condensation
The preparation method of bacillus ACCC10229 bacterium solution preferably include successively to carry out bacillus coagulans ACCC10229 activation and
Expand culture to obtain.In the present invention, the mode of the activation is preferably by bacillus coagulans ACCC10229 in MRS solid
It crosses on slant medium, by scribed culture medium in 35 DEG C of cultures to growing bacterium colony.In the present invention, time of the activation
Number preferably 2 times.In the present invention, the single colonie after activation is preferably inoculated in MRS liquid by the mode for expanding culture
In culture medium, at 35 DEG C, 120rpm overnight incubation obtains bacillus coagulans ACCC10229 bacterium solution.
In the present invention, the temperature of the fermentation is preferably 30~40 DEG C, and more preferably 35 DEG C；The stirring speed of the fermentation
Degree is preferably 80~120rpm, more preferably 100rpm；The time of the fermentation is preferably 20~28h, more preferably for 24 hours.
In the present invention, bacillus coagulans ACCC10229 is homofermentative lactic bacteria, and condensation is added in ensilage
After bacillus ACCC10229 fermentation liquid, homofermentative lactic is carried out, increases fermenting speed, lactic acid content increases, homolactic
Bacterium occupies advantage, inhibits the growth of clostridium butyricum and enterobacteria, so that butyric acid and acetic acid content be made to decline, reduces in alfalfa ensilage
The content of butyric acid.
In the present invention, the preparation method of the alfalfa ensilage, preferably includes following steps:
1) bacillus coagulans ACCC10229 is fermented, obtains bacillus coagulans ACCC10229 fermentation liquid；
2) clover is mixed with the bacillus coagulans ACCC10229 fermentation liquid of the step 1), obtains total mixture；
3) total mixture of the step 2) is sealed 40~50d of fermentation, obtains alfalfa ensilage.
The present invention ferments bacillus coagulans ACCC10229, obtains bacillus coagulans ACCC10229 fermentation
Liquid.In the present invention, the fermentation process of the bacillus coagulans ACCC10229 is preferably condensation gemma described in above scheme
The preparation method of bacillus ACCC10229 fermentation liquid.
After obtaining bacillus coagulans ACCC10229 fermentation liquid, the present invention is preferably by clover and bacillus coagulans
The mixing of ACCC10229 fermentation liquid, obtains total mixture.In the present invention, clover is preferably cut into 2~5cm's before the mixing
Clover section, more preferably 4cm.In the present invention, the quality volume of the clover and bacillus coagulans ACCC10229 fermentation liquid
Than being preferably 1Kg:1.4~1.6mL, more preferably 1Kg:1.5mL.
The present invention is not particularly limited the source of clover, using this field routine clover.In the embodiment of the present invention
The polar bear kind provided with Dan Nong seeds company, " the beach saline land greening soil of agricultural resource and Environmental Research Institute in Tianjin
Earth improvement scientific research and testing base " plantation obtains.
In the present invention, preferably alfalfa ensilage leavening is diluted before the mixing；The diluted multiple is preferred
It is 3~5 times, more preferably 4 times.The dilution is preferably water with dilution.In the present invention, the mixed mode is preferably
Alfalfa ensilage leavening is sprayed on clover.
After obtaining total mixture, the total mixture is preferably sealed 40~50d of fermentation by the present invention, obtains alfalfa ensilage.
In the present invention, the mode of the sealing is preferably sealed using vinyon.Preferred discharge air before sealing.?
In the present invention, the temperature of the sealing and fermenting is preferably 20~30 DEG C, and more preferably 25 DEG C.The time of the sealing and fermenting is preferred
For 45d.
In order to further illustrate the present invention, technical solution provided by the invention is retouched in detail below with reference to embodiment
It states, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
Bacillus coagulans ACCC10229 is taken out from cryopreservation tube with sterilized oese, on MRS solid slope strokes
Scribed MRS solid medium is placed in 35 DEG C of incubators and turns out bacterium colony by line, and picking single colonie is connected to after activation 2 times
In 35 DEG C in MRS fluid nutrient medium, 120rpm is overnight, and it is spare to obtain bacillus coagulans ACCC10229 bacterium solution.
It is 1.0 × 10 that obtained bacillus coagulans ACCC10229 bacterium solution, which is adjusted to bacteria concentration,9After CFU/g, inoculation
(inoculum concentration is the 10% of fluid nutrient medium weight) (peptone 12g/L, powdered beef 12g/L, yeast powder into fluid nutrient medium
6g/L, glucose 24g/L, dipotassium hydrogen phosphate 2g/L, diammonium hydrogen citrate 2g/L, sodium acetate 5g/L, magnesium sulfate 0.58g/L, sulphur
Sour manganese 0.25g/L, tween 5mL, distilled water constant volume to 1L；After preparing liquid culture medium by above-mentioned formula, adjust pH value to
5.9) it is cultivated, at 35 DEG C, under conditions of 100rpm for 24 hours, obtains bacillus coagulans ACCC10229 fermentation liquid.
The test polar bear kind that clover is that Dan Nong seeds company provides, planting area is Agriculture In Tianjin resource and ring
" beach saline land green soil improves scientific research and testing base " of border research institute, mu application rate 1.2Kg.Clover flower at the beginning of first batch
Phase cradles clover and is cut to the clover of 2~5cm with hay cutter after sunning for 24 hours (dry matter content 29.13 ± 0.75%)
Section, for making alfalfa ensilage.
Embodiment 2
The clover section of Example 1, the bacillus coagulans ACCC10229 fermentation liquid that embodiment 1 is prepared spray
In clover section, above (mass volume ratio of clover section and bacillus coagulans ACCC10229 fermentation liquid is 1Kg:1.6mL, to guarantee
The homogeneity of alfalfa ensilage addition is sprayed onto clover section again after bacillus coagulans ACCC10229 fermentation liquid is first diluted 3 times
On), be packed into polyethylene plastic bag (25cm × 35cm), 3 repetitions are arranged in every bag of 300g, carried out simultaneously with sealing machine pumping and
Bag is opened in sealing after 20 DEG C of fermentation 50d, analyzes alfalfa ensilage ingredient.
Embodiment 3
The clover section of Example 1, the bacillus coagulans ACCC10229 fermentation liquid that embodiment 1 is prepared spray
In clover section, above (mass volume ratio of clover section and alfalfa ensilage leavening is 1Kg:1.4mL, to guarantee alfalfa ensilage addition
Homogeneity is sprayed on clover segment again after bacillus coagulans ACCC10229 fermentation liquid is first diluted 5 times), it is packed into polyethylene
3 repetitions are arranged in polybag (25cm × 35cm), every bag of 300g, are evacuated and are sealed simultaneously with sealing machine, fermented at 30 DEG C
Bag is opened after 40d, analyzes alfalfa ensilage ingredient.
Embodiment 4
The clover section of Example 1, the bacillus coagulans ACCC10229 fermentation liquid that embodiment 1 is prepared spray
In clover section, above (mass volume ratio of clover section and alfalfa ensilage leavening is 1Kg:1.5mL, to guarantee alfalfa ensilage addition
Homogeneity is sprayed onto again in clover section after bacillus coagulans ACCC10229 fermentation liquid is first diluted 4 times), it is packed into polyethylene modeling
3 repetitions are arranged in material bag (25cm × 35cm), every bag of 300g, are evacuated and are sealed simultaneously with sealing machine, fermented at 25 DEG C
Bag is opened after 45d, analyzes alfalfa ensilage ingredient.
Comparative example 1
In addition to not adding fermentation liquid, other operating procedures are identical with embodiment 4, alfalfa ensilage prepared, as control
Group 1.
Using the activation of embodiment 1, expansion culture and fermentation process preparation Pediococcus acidilactici CGMCC1.4, (purchase is certainly Chinese
General Microbiological Culture preservation administrative center) fermentation liquid, the above-mentioned lactic acid sheet ball being prepared is added when preparing alfalfa ensilage
Bacterium CGMCC1.4 fermentation liquid, in addition to fermentation liquid is different, other operating procedures are identical with embodiment 4, prepare alfalfa ensilage, make
For control group 2.
Preparing lactobacillus plantarum CICC20765 using the activation of embodiment 1, expansion culture and fermentation process, (purchase is in
State's Research for Industrial Microbial Germ preservation administrative center) fermentation liquid, the lactobacillus plantarum CICC20765 fermentation liquid and reality that will be prepared
After the bacillus coagulans ACCC10229 fermentation liquid of example 1 is applied according to the mass ratio mixing of 1:1, it is added during the fermentation,
In addition to fermentation liquid is different, other operating procedures are identical with embodiment 4, prepare alfalfa ensilage, and as a control group 3.
Using the activation of embodiment 1, expand culture and fermentation process preparation Pediococcus pentosaceus CICC22737 (in purchase certainly
State's Research for Industrial Microbial Germ preservation administrative center) fermentation liquid, the Pediococcus pentosaceus CICC22737 fermentation liquid and reality that will be prepared
After the bacillus coagulans ACCC10229 fermentation liquid of example 1 is applied according to the mass ratio mixing of 1:1, it is added during the fermentation,
In addition to fermentation liquid is different, other operating procedures are identical with embodiment 4, prepare alfalfa ensilage, and as a control group 4.
Embodiment 5
By the forage analysis China services center CVAS by near infrared spectrum (NearInfrared, NIR) to embodiment 4 and
The alfalfa ensilage ingredient that control group 1~4 is prepared in comparative example 1 is analyzed, and specific measurement result is as shown in table 1.
Influence of the 1 different fermentations agent of table to component content in alfalfa ensilage
As can be seen from Table 1, butyric acid in alfalfa ensilage can be significantly reduced in bacillus coagulans ACCC10229 fermentation liquid
Content reduces by 73.64%~82.26% relative to control group 1~4.Meanwhile lactic acid content is also relatively high；Ammoniacal nitrogen, pH and
Acetic acid content is relatively low.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. bacillus coagulans ACCC10229 is reducing the application in alfalfa ensilage butyric acid content.
2. application according to claim 1, which is characterized in that the bacillus coagulans ACCC10229 ferments
Afterwards, obtained fermentation liquid is added in clover and prepares alfalfa ensilage.
3. application according to claim 2, which is characterized in that the system of the bacillus coagulans ACCC10229 fermentation liquid
Preparation Method includes: that bacillus coagulans ACCC10229 bacterium solution is inoculated into liquid state fermentation culture medium, at 30~40 DEG C, 80~
Ferment 20~28h under conditions of 120rpm, obtains bacillus coagulans ACCC10229 fermentation liquid；The liquid state fermentation culture medium
It takes water as a solvent, the component including following concentration: peptone 12g/L, powdered beef 12g/L, yeast powder 6g/L, glucose 24g/L,
Dipotassium hydrogen phosphate 2g/L, diammonium hydrogen citrate 2g/L, sodium acetate 5g/L, magnesium sulfate 0.58g/L, manganese sulfate 0.25g/L and tween
5mL/L, the pH value of the liquid state fermentation culture medium are 5.8~6.0.
4. application according to claim 3, which is characterized in that the viable bacteria of the bacillus coagulans ACCC10229 bacterium solution
Number is 1.0 × 108~1.0 × 1010CFU/g。
5. application according to claim 3, which is characterized in that the inoculum concentration is liquid state fermentation total weight of medium
5%~15%.
6. application described in any one according to claim 1~5, which is characterized in that the preparation method of the alfalfa ensilage, packet
Include following steps:
1) bacillus coagulans ACCC10229 is fermented, obtains bacillus coagulans ACCC10229 fermentation liquid；
2) clover is mixed with the bacillus coagulans ACCC10229 fermentation liquid of the step 1), obtains total mixture；
3) total mixture of the step 2) is sealed 40~50d of fermentation, obtains alfalfa ensilage.
7. application according to claim 6, which is characterized in that clover and bacillus coagulans in the step 2)
The mass volume ratio of ACCC10229 fermentation liquid is 1Kg:1.4~1.6mL.
8. application according to claim 6, which is characterized in that further include that will condense gemma bar before being mixed described in step 2)
Bacterium ACCC10229 fermentation liquid is diluted；The diluted multiple is 3~5 times.
9. application according to claim 6, which is characterized in that further include being cut into clover before being mixed described in step 2)
The clover section of 2~5cm.
10. application according to claim 6, which is characterized in that the temperature being sealed by fermentation in the step 3) is 20~30
℃。
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2019
- 2019-01-30 CN CN201910089425.7A patent/CN109619289A/en active Pending
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| CN103160455A (en) * | 2013-03-27 | 2013-06-19 | 杭州保安康生物技术有限公司 | Preparation method of spore preparation of bacillus coagulans |
| CN105567585A (en) * | 2015-12-16 | 2016-05-11 | 天津市畜牧兽医研究所 | Compound microbial agent for silage and application thereof in whole-plant corn silage |
| CN106819407A (en) * | 2017-04-07 | 2017-06-13 | 天津市畜牧兽医研究所 | A kind of additives for ensiling and its application for alfalfa ensilage |
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