CN109929771A - Ensiling agent and preparation method thereof - Google Patents
Ensiling agent and preparation method thereof Download PDFInfo
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- CN109929771A CN109929771A CN201811353692.2A CN201811353692A CN109929771A CN 109929771 A CN109929771 A CN 109929771A CN 201811353692 A CN201811353692 A CN 201811353692A CN 109929771 A CN109929771 A CN 109929771A
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Abstract
The invention discloses a kind of ensiling agent and preparation method thereof, which includes the component of following mass fraction: 2~4 parts of lactobacillus plantarum, 2~4 parts of lactobacillus acidophilus, 1~3 part of Pediococcus acidilactici, 1~3 part of lactobacillus reuteri, 1 part of lactobacillus buchneri, 1 part of bacillus subtilis, 1 part of cellulase.The present invention passes through screening efficient lactic acid bacterium, carry out scientific compatibility, successfully develop ensiling agent, it is operated under working condition in normal ensiling, realize the pH value for quickly reducing ensiling object, inhibit acetic acid bacteria, saccharomycete, enterobacteria, clostridium, bacillus, mould and Li bacillus, produces the high-grade maize complete stool ensilage of high value.
Description
Technical field
The present invention relates to a kind of ensiling agent and preparation method thereof.
Background technique
Ensilage is the important roughage source of milk cow, in ensiling manufacturing process, although ensure that moisture, sugar
Point and anaerobic condition after, ensiling still may will fail, and main cause is the lactic acid grown nonparasitically upon another plant naturally in plant with environmental pollution
Bacterium type and quantity are widely different, and targeted content of lactic acid bacteria is relatively low, can not sugared part in Rapid Fermentation ensiling object, fastly
Prompt drop low ph value, to inhibit the growth and breeding of other miscellaneous bacterias.The failure of ensiling production not only will lead to feed economic loss, go back
It will cause the pollution of environment.
Summary of the invention
In order to overcome drawbacks described above, the present invention provides a kind of ensiling agent and preparation method thereof, can be in good ensiling pipe
On the basis of reason measure, Silage Quality is further promoted, reduces the economic loss of ensiling failure.
The present invention is to solve technical solution used by its technical problem: a kind of ensiling agent, including following mass parts
Several component: 2~4 parts of lactobacillus plantarum, 2~4 parts of lactobacillus acidophilus, 1~3 part of Pediococcus acidilactici, lactobacillus reuteri 1~3
Part, 1 part of lactobacillus buchneri, 1 part of bacillus subtilis, 1 part of cellulase.
The present invention also provides a kind of preparation methods of ensiling agent as described above, comprising the following steps:
Step 1, matrix manufacturing is cultivated:
Firstly, by peptone 5kg, glucose 7kg, yeast powder 2kg, sodium acetate 1kg, precipitated calcium carbonate 2kg, ammonium citrate
0.4kg, tween 0.2kg, additive package 50g, with distilled water dosage at 200L, stirring and dissolving is simultaneously heated to 65~75 DEG C；So
Afterwards, with 20% sodium hydroxide (NaOH) solution tune pH value for 7.0~7.2, ingredient is completed；It is sterilized 15 minutes with 115~120 DEG C again
Afterwards, 37 DEG C are cooled the temperature to, sterilising medium is obtained；
Step 2, the activation and expansion culture of strain:
The lactobacillus plantarum of refrigeration is inoculated in the above-mentioned sterilising medium of 100ml once to be activated, temperature is 37~40
DEG C, it being detected after 30 hours, by 100ml bacterium solution, all the above-mentioned sterilising medium of access 500ml carries out re-activation after detection is up to standard,
Temperature be 37~40 DEG C, 30 hours after detect it is up to standard after expand culture 72 hours；
The activation and expansion culture of lactobacillus acidophilus are the same as above-mentioned lactobacillus plantarum；
The activation and expansion culture of Pediococcus acidilactici are the same as above-mentioned lactobacillus plantarum；
The activation and expansion culture of lactobacillus reuteri are the same as above-mentioned lactobacillus plantarum；
The activation and expansion culture of lactobacillus buchneri are the same as the above-mentioned object lactobacillus of plant；
The activation and expansion culture of bacillus subtilis are the same as above-mentioned lactobacillus plantarum；
Step 3, the collection of bacterium mud:
The lactobacillus plantarum liquid after fermentation is centrifuged with centrifuge, and collects bacterium mud；
The bacterium mud collection method of lactobacillus acidophilus is the same as above-mentioned lactobacillus plantarum；
The bacterium mud collection method of Pediococcus acidilactici is the same as above-mentioned lactobacillus plantarum；
The above-mentioned same lactobacillus plantarum of the bacterium mud collection method of lactobacillus reuteri；
The bacterium mud collection method of lactobacillus buchneri is the same as above-mentioned lactobacillus plantarum；
The bacterium mud collection method of bacillus subtilis is the same as above-mentioned lactobacillus plantarum；
Step 4, the freeze-drying of bacterium mud:
Freeze-drying preparation is carried out to be coated with bacterium powder treatment process for lactobacillus plantarum mud collected by step 3, it is specific to walk
Suddenly are as follows: protective agent is added in a. bacterium mud, and protectant mass fraction reaches 25%, mixes, and b., which is contained, to be freezed into freezing plate at -30 DEG C
50 minutes, c. was freezed 60 minutes at -80 DEG C, and d. is freeze-dried 13~15 hours on frozen vacuum dryer, and e. freeze-dried powder is close
It closes and saves in -4 DEG C of refrigerators；
The bacterium mud freeze-drying method of lactobacillus acidophilus is the same as above-mentioned lactobacillus plantarum；
The bacterium mud freeze-drying method of Pediococcus acidilactici is the same as above-mentioned lactobacillus plantarum；
The bacterium mud freeze-drying method of lactobacillus reuteri is the same as above-mentioned lactobacillus plantarum；
The bacterium mud freeze-drying method of lactobacillus buchneri is the same as above-mentioned lactobacillus plantarum；
Bacillus subtilis bacterium mud collected by step 3 is subjected to freeze-drying preparation, specific steps are as follows: a. contains flat into freezing
Ware freezes 50 minutes at -30 DEG C, and b. is freezed 60 minutes at -80 DEG C, and it is small that c. is freeze-dried 13~15 on frozen vacuum dryer
When, in d. freeze-dried powder -4 DEG C of refrigerators of closed preservation；
Step 5, prepared by finished product:
Take 2~4 parts of 2~4 parts of lactobacillus plantarum freeze-dried powder, lactobacillus acidophilus freeze-drying powder, the lactic acid sheet ball prepared in step 4
1~3 part of bacterium freeze-dried powder, 1~3 part of lactobacillus reuteri freeze-dried powder, 1 part of lactobacillus buchneri freeze-dried powder, bacillus subtilis freeze-drying
1 part of powder, 1 part of cellulase pack after mixing under aseptic conditions, obtain ensiling agent.
As a further improvement of the present invention, the additive package in the step 1 by following mass fraction substance group
At: 3 parts of dipotassium hydrogen phosphate, 6 parts of magnesium sulfate, 2 parts of manganese sulfate.
As a further improvement of the present invention, the protective agent in the step 4 includes the ingredient of following mass percentage:
Sucrose 40%, skimmed milk 40%, concentration of sodium glutamate 20%.
The beneficial effects of the present invention are: the present invention carries out scientific compatibility, successfully develops blueness by screening efficient lactic acid bacterium
Storage agent realizes the pH value for quickly reducing ensiling object, inhibits acetic acid bacteria, saccharomycete, intestines in the case where normal ensiling operates working condition
Bacillus, clostridium, bacillus, mould and Li bacillus produce the high-grade maize complete stool ensilage of high value.
Specific embodiment
With reference to embodiments, it elaborates to the present invention.But protection scope of the present invention is not limited to following embodiments,
I.e. in every case with simple equivalent changes and modifications made by scope of the present invention patent and description, all still belong to the present invention
Within patent covering scope.
A kind of ensiling agent, the component including following mass fraction: 2~4 parts of lactobacillus plantarum, 2~4 parts of lactobacillus acidophilus,
1~3 part of Pediococcus acidilactici, 1~3 part of lactobacillus reuteri, 1 part of lactobacillus buchneri, 1 part of bacillus subtilis, cellulase 1
Part.
The preparation method of above-mentioned ensiling agent, comprising the following steps:
Step 1, matrix manufacturing is cultivated:
Firstly, by peptone 5kg, glucose 7kg, yeast powder 2kg, sodium acetate 1kg, precipitated calcium carbonate 2kg, ammonium citrate
0.4kg, tween 0.2kg, additive package (dipotassium hydrogen phosphate 15g, magnesium sulfate 30g, manganese sulfate 10g) 55g, with distilled water dosage
At 200L, stirring and dissolving is simultaneously heated to 65~75 DEG C；Then, with 20% sodium hydroxide (NaOH) solution tune pH value be 7.0~
7.2, complete ingredient；After sterilizing 15 minutes with 115~120 DEG C again, 37 DEG C are cooled the temperature to, obtains sterilising medium；
Step 2, the activation and expansion culture of strain:
The lactobacillus plantarum of refrigeration is inoculated in the above-mentioned sterilising medium of 100ml once to be activated, temperature is 37~40
DEG C, it being detected after 30 hours, by 100ml bacterium solution, all the above-mentioned sterilising medium of access 500ml carries out re-activation after detection is up to standard,
Temperature be 37~40 DEG C, 30 hours after detect it is up to standard after expand culture 72 hours；
The activation and expansion culture of lactobacillus acidophilus are the same as above-mentioned lactobacillus plantarum；
The activation and expansion culture of Pediococcus acidilactici are the same as above-mentioned lactobacillus plantarum；
The activation and expansion culture of lactobacillus reuteri are the same as above-mentioned lactobacillus plantarum；
The activation and expansion culture of lactobacillus buchneri are the same as above-mentioned lactobacillus plantarum；
The activation and expansion culture of bacillus subtilis are the same as above-mentioned lactobacillus plantarum；
Step 3, the collection of bacterium mud:
The lactobacillus plantarum liquid after fermentation is centrifuged with centrifuge, and collects bacterium mud；
The bacterium mud collection method of lactobacillus acidophilus is the same as above-mentioned lactobacillus plantarum；
The bacterium mud collection method of Pediococcus acidilactici is the same as above-mentioned lactobacillus plantarum；
The bacterium mud collection method of lactobacillus reuteri is the same as above-mentioned lactobacillus plantarum；
The bacterium mud collection method of lactobacillus buchneri is the same as above-mentioned lactobacillus plantarum；
The bacterium mud collection method of bacillus subtilis is the same as above-mentioned lactobacillus plantarum；
Step 4, the freeze-drying of bacterium mud:
Freeze-drying preparation is carried out to be coated with bacterium powder treatment process for lactobacillus plantarum mud collected by step 3, it is specific to walk
Suddenly are as follows: protective agent is added in a. bacterium mud, and protectant mass fraction reaches 25%, mixes, and b., which is contained, to be freezed into freezing plate at -30 DEG C
50 minutes, c. was freezed 60 minutes at -80 DEG C, and d. is freeze-dried 13~15 hours on frozen vacuum dryer, and e. freeze-dried powder is close
It closes and saves in -4 DEG C of refrigerators；
The bacterium mud freeze-drying method of lactobacillus acidophilus is the same as above-mentioned lactobacillus plantarum；
The bacterium mud freeze-drying method of Pediococcus acidilactici is the same as above-mentioned lactobacillus plantarum；
The above-mentioned same lactobacillus plantarum of the bacterium mud freeze-drying method of lactobacillus reuteri；
The bacterium mud freeze-drying method of lactobacillus buchneri is the same as above-mentioned lactobacillus plantarum；
Bacillus subtilis bacterium mud collected by step 3 is subjected to freeze-drying preparation, specific steps are as follows: a. contains flat into freezing
Ware freezes 50 minutes at -30 DEG C, and b. is freezed 60 minutes at -80 DEG C, and it is small that c. is freeze-dried 13~15 on frozen vacuum dryer
When, in d. freeze-dried powder -4 DEG C of refrigerators of closed preservation；
Step 5, prepared by finished product:
Take 2~4 parts of 2~4 parts of lactobacillus plantarum freeze-dried powder, lactobacillus acidophilus freeze-drying powder, the lactic acid sheet ball prepared in step 4
1~3 part of bacterium freeze-dried powder, 1~3 part of lactobacillus reuteri freeze-dried powder, 1 part of lactobacillus buchneri freeze-dried powder, bacillus subtilis freeze-drying
1 part of powder, 1 part of cellulase pack after mixing under aseptic conditions, obtain ensiling agent.
Action principle of the invention under anaerobic, utilizes lactobacillus plantarum, lactobacillus acidophilus, lactic acid for ensiling raw material
Piece coccus, lactobacillus reuteri fermentation, convert lactic acid for plant soluble-carbohydrate, so that the pH value of ensiling raw material is fast
Prompt drop to 4.2 hereinafter, reach inhibit acetic acid bacteria, saccharomycete, enterobacteria, clostridium, bacillus, mould and Li bacillus etc. not by
The growth and breeding of microorganism is welcome, crop nutritive peculiarity is kept, produces the quality silage of high value.Below in conjunction with implementation
Example elaborates to effect of the invention.
1, experimental material
1) test site: Tianjin Feng Tai agriculture and animal husbandry Science and Technology Ltd., the Heilungkiang Nenjiang County pasture Jian Ye.
2) ensiling raw material: using whole-plant corn as ensiling raw material, guarantee the water content of raw material 60%~70%.
3) additive amount: one ton of ensiling raw material of fermentation adds 5 grams of this ensiling agent.
2, experimental design
Experiment sets control group (natural ensiling: not adding ensiling agent), experimental group (addition ensiling agent of the present invention), and experimental group is
Following Examples 1 to 5.Wherein, Examples 1 to 5 is respectively according to composition of raw materials shown in table 1 and technique of the present invention step
It is rapid to obtain.
1. Examples 1 to 5 of table
3, experimental method
1) for corn silage Cutting Height other than 15 centimetres, Cutting Length is 2cm or so, enters cellar compacting, ensilage is discharged
In air, compacted density be not less than 750kg/m3。
2) pH: behind Kaifeng, taking ensilage, is measured with pH electrode test table.
Dry matter (DM) content: using 65 DEG C of seasoning measurements.
Crude protein (CP) content: it is measured using GB6432-86.
Ammoniacal nitrogen (TBN) content: it is measured using direct distillation.
Cellulose (neutral detergent fiber NDF, acidity wash silk ribbon fiber ADF and acidic cleaning lignin ADL) content: model is used
The measurement of family name's Fiber Assay.
Organic acid content: it is measured using efficient liquid phase for spectrometry.
3) V-Score standards of grading (2001), Kaiser method score-system can be used in wilted grass silage appraisement system
(2005) and 1996 tentative standard of the Ministry of Agriculture provides.
Corn silage appraisement system of the present invention presses the silage fermentation flavor evaluation that Kaiser method score-system (2005) proposes
Standard.Scoring rank divides Pyatyi from high to low: 1 grade, 2 grades, 3 grades, 4 grades, 5 grades.
4, experimental result
Embodiment 1: pH value, organic acid content and the scoring of different ensilages are shown in Table 2, different ensilage chemical components
Mass fraction be shown in Table 3.
Embodiment 2: pH value, organic acid content and the scoring of different ensilages are shown in Table 4, different ensilage chemical components
Mass fraction be shown in Table 5.
Embodiment 3: pH value, organic acid content and the scoring of different ensilages are shown in Table 6, different ensilage chemical components
Mass fraction be shown in Table 7.
Embodiment 4: pH value, organic acid content and the scoring of different ensilages are shown in Table 8, different ensilage chemical components
Mass fraction be shown in Table 9.
Embodiment 5: pH value, organic acid content and the scoring of different ensilages are shown in Table 10, different ensilage chemistry at
The mass fraction divided is shown in Table 11.
2. embodiment 1 of table: pH value, organic acid content and the scoring of different ensilages
3. embodiment 1 of table: the mass fraction of different ensilage chemical components
4. embodiment 2 of table: pH value, organic acid content and the scoring of different ensilages
5. embodiment 2 of table: the mass fraction of different ensilage chemical components
6. embodiment 3 of table: pH value, organic acid content and the scoring of different ensilages
7. embodiment 3 of table: the mass fraction of different ensilage chemical components
8. embodiment 4 of table: pH value, organic acid content and the scoring of different ensilages
9. embodiment 4 of table: the mass fraction of different ensilage chemical components
10. embodiment 5 of table: pH value, organic acid content and the scoring of different ensilages
11. embodiment 5 of table: the mass fraction of different ensilage chemical components
6, experiment conclusion
By the experimental data of embodiment 1, embodiment 2, embodiment 3, embodiment 4 and embodiment 5 it can be seen that right
According in group, ensilage sensory assessment is up to 90 points (1 grades), and worst is 70 points (3 grades)；In experimental group, ensilage quality
Scoring is 100 points (1 grades).Compared to the control group, ensilage chemical component mass fraction also significantly improves experimental group.
In conclusion the present invention in the case where normal ensiling operates working condition, realizes the pH value for quickly reducing ensiling object, suppression
Acetic acid bacteria, saccharomycete, enterobacteria, clostridium, bacillus, mould and Li bacillus processed, the high-quality ensiling for producing high value are raised
Material.
Claims (4)
1. a kind of ensiling agent, which is characterized in that the component including following mass fraction: 2~4 parts of lactobacillus plantarum, acidophilus cream bar
2~4 parts of bacterium, 1~3 part of Pediococcus acidilactici, 1~3 part of lactobacillus reuteri, 1 part of lactobacillus buchneri, 1 part of bacillus subtilis, fibre
Tie up plain 1 part of enzyme.
2. a kind of preparation method of ensiling agent as described in claim 1, which comprises the following steps:
Step 1, matrix manufacturing is cultivated:
Firstly, by peptone 5kg, glucose 7kg, yeast powder 2kg, sodium acetate 1kg, precipitated calcium carbonate 2kg, ammonium citrate
0.4kg, tween 0.2kg, additive package 50g, with distilled water dosage at 200L, stirring and dissolving is simultaneously heated to 65~75 DEG C；So
Afterwards, with 20% sodium hydroxide (NaOH) solution tune pH value for 7.0~7.2, ingredient is completed；It is sterilized 15 minutes with 115~120 DEG C again
Afterwards, 37 DEG C are cooled the temperature to, sterilising medium is obtained；
Step 2, the activation and expansion culture of strain:
The lactobacillus plantarum of refrigeration is inoculated in the above-mentioned sterilising medium of 100ml once to be activated, temperature be 37~40 DEG C, 30
It is detected after hour, by 100ml bacterium solution, all the above-mentioned sterilising medium of access 500ml carries out re-activation, temperature after detection is up to standard
Detected after being 37~40 DEG C, 30 hours it is up to standard after expand culture 72 hours；
The activation and expansion culture of lactobacillus acidophilus are the same as above-mentioned lactobacillus plantarum；
The activation and expansion culture of Pediococcus acidilactici are the same as above-mentioned lactobacillus plantarum；
The activation and expansion culture of lactobacillus reuteri are the same as above-mentioned lactobacillus plantarum；
The activation and expansion culture of lactobacillus buchneri are the same as the above-mentioned object lactobacillus of plant；
The activation and expansion culture of bacillus subtilis are the same as above-mentioned lactobacillus plantarum；
Step 3, the collection of bacterium mud:
The lactobacillus plantarum liquid after fermentation is centrifuged with centrifuge, and collects bacterium mud；
The bacterium mud collection method of lactobacillus acidophilus is the same as above-mentioned lactobacillus plantarum；
The bacterium mud collection method of Pediococcus acidilactici is the same as above-mentioned lactobacillus plantarum；
The above-mentioned same lactobacillus plantarum of the bacterium mud collection method of lactobacillus reuteri；
The bacterium mud collection method of lactobacillus buchneri is the same as above-mentioned lactobacillus plantarum；
The bacterium mud collection method of bacillus subtilis is the same as above-mentioned lactobacillus plantarum；
Step 4, the freeze-drying of bacterium mud:
Freeze-drying preparation, specific steps are carried out to be coated with bacterium powder treatment process for lactobacillus plantarum mud collected by step 3 are as follows:
A. protective agent is added in bacterium mud, and protectant mass fraction reaches 25%, mixes, and b., which is contained, freezes 50 points at -30 DEG C into freezing plate
Clock, c. are freezed 60 minutes at -80 DEG C, and d. is freeze-dried 13~15 hours on frozen vacuum dryer, the closed guarantor of e. freeze-dried powder
It deposits in -4 DEG C of refrigerators；
The bacterium mud freeze-drying method of lactobacillus acidophilus is the same as above-mentioned lactobacillus plantarum；
The bacterium mud freeze-drying method of Pediococcus acidilactici is the same as above-mentioned lactobacillus plantarum；
The bacterium mud freeze-drying method of lactobacillus reuteri is the same as above-mentioned lactobacillus plantarum；
The bacterium mud freeze-drying method of lactobacillus buchneri is the same as above-mentioned lactobacillus plantarum；
Bacillus subtilis bacterium mud collected by step 3 is subjected to freeze-drying preparation, specific steps are as follows: a. is contained into freezing plate
It being freezed 50 minutes at -30 DEG C, b. is freezed 60 minutes at -80 DEG C, and c. is freeze-dried 13~15 hours on frozen vacuum dryer,
D. in -4 DEG C of refrigerators of the closed preservation of freeze-dried powder；
Step 5, prepared by finished product:
2~4 parts of 2~4 parts of lactobacillus plantarum freeze-dried powder, lactobacillus acidophilus freeze-drying powder preparing in step 4, Pediococcus acidilactici is taken to freeze
1~3 part of dry powder, 1~3 part of lactobacillus reuteri freeze-dried powder, 1 part of lactobacillus buchneri freeze-dried powder, bacillus subtilis freeze-dried powder 1
Part, 1 part of cellulase pack after mixing under aseptic conditions, obtain ensiling agent.
3. preparation method according to claim 2, it is characterised in that: the additive package in the step 1 is by following matter
The material composition of amount number: 3 parts of dipotassium hydrogen phosphate, 6 parts of magnesium sulfate, 2 parts of manganese sulfate.
4. preparation method according to claim 2, it is characterised in that: the protective agent in the step 4 includes following quality
The ingredient of percentage composition: sucrose 40%, skimmed milk 40%, concentration of sodium glutamate 20%.
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Cited By (1)
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| CN115669810A (en) * | 2022-10-17 | 2023-02-03 | 湖南农业大学 | Rice silage additive |
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Application publication date: 20190625 |