CN101869230B - Microbial additive for alfalfa haylage, preparation method and application thereof - Google Patents
Microbial additive for alfalfa haylage, preparation method and application thereof Download PDFInfo
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- CN101869230B CN101869230B CN 201010224736 CN201010224736A CN101869230B CN 101869230 B CN101869230 B CN 101869230B CN 201010224736 CN201010224736 CN 201010224736 CN 201010224736 A CN201010224736 A CN 201010224736A CN 101869230 B CN101869230 B CN 101869230B
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- aspergillus oryzae
- streptococcus lactis
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Abstract
The invention discloses a microbial additive for alfalfa haylage, a preparation method and an application thereof. The additive is prepared from a bacterial suspension of aspergillusoryzae and an MRS liquid nutrient medium for streptococcus lactis according to the ratio of 50-70%: 50-30% in weight percent, wherein per milliliter of the bacterial suspension of the aspergillusoryzae at least contains 1.0*108 aspergillusoryzae spores, and per milliliter of the MRS liquid nutrient medium at least contains 4.0*108 cfu streptococcus lactis. The microbial additive quickens the initial fermentation rate of lactobacillus, enhances acid production capability, effectively inhibits the activity of saccharomycetes and other undesirable microorganism, strengths haylage effect and effectively promotes the production performance of cows, beef cattle, mutton sheep and swine. The strain adopted by the invention can be obtained easily, can be produced massively on application and has practicability.
Description
Technical field
The present invention relates to a kind of microbe additive, specifically relate to the half-dried forage ensiling of a kind of clover microbe additive.
Background technology
Clover is a kind of Perennial legume forages, and nutritive value is abundant, is the forage grass of plant-eating animal high-quality, and the good reputation of " King of Pasture " is arranged.For a long time, the major product of ALFALFA PRODUCTION is hay, but in the modulated process of the many local alfalfa hay of China, all exist drench with rain, the loss such as fallen leaves, the General Loss rate is about 30%.Particularly in the main producing region of China clover, because rainwater and heat same period, the loss late that the clover harvest season is drenched with rain is higher.And northern China clover main producing region cradles 2,3 batches clover and is difficult to shine high-quality hay processed, adopt the way production dehydrated alfalfa of oven dry affected by rainy season, can produce high-quality grass product, but drying plant is expensive, and consume mass energy, can only be in limited range applications.For this reason, adopting alfalfa ensilage is to address the above problem comparatively desirable measure.
Alfalfa ensilage not only can reduce nutrient loss, and can keep the nutritive peculiarity of green forage.Good palatability behind the alfalfa ensilage, quality better, digestibility be high, can long preservation, and wrap up in ensiling that packet form the stores field that also can circulate with stretched film, can sell in the strange land and utilize.Because the clover soluble sugar content is low, buffer capacity is high, belongs to the leguminous forage of difficult ensiling, and directly silage effect is bad, need to use additive in ensilage.At present the domestic half-dried bale of clover is wrapped up in and often in the bag ensilage is added lactic acid bacteria, and its defective is wrapped up in for bundle residually in the bag has an air, and lactic acid bacteria can't normal growth, causes the mould Fast-propagation, causes the Alfalfa Silage nutrient loss, in addition mould lose rotten.For improving the quality of alfalfa haylage, be necessary to improve on present addictive for alfalfa silage basis, wrap up in oxygen in the bag to consume the half-dried bale of Alfalfa Silage, the initial fermenting speed of lactic acid bacteria is accelerated, strengthen acid producing ability, thereby effectively suppress the activity of saccharomycete and other undesirable microorganism, make the alfalfa haylage process reach as early as possible the condition of anaerobic environment, guarantee to preserve to greatest extent the nutritive value of clover.
Summary of the invention
The present invention is directed in the prior art can't deaeration defective and deficiency, a kind of microbial additive for alfalfa haylage is provided, this microbe additive can make organic acid content increase, pH value descend, suppress harmful fungus growth, promote lactobacter growth, reached the purpose of fast reducing pH and raising ensiling quality.
The present invention is achieved by the following measures:
The present invention is directed to clover forage Air affects the deficiency of lactic acid bacteria normal growth too much, select to have done certain adjustment at bacterial classification, in streptococcus lactis, add aerobic bacteria---aspergillus oryzae, strengthened silage effect, increased the production performance of milk cow, beef cattle, mutton sheep.The aspergillus oryzae spore germination can consume rapidly oxygen, provides the anaerobic environment that is beneficial to lactobacter growth, makes the breeding of lactic acid bacteria rapid, high volume.This additive can descend organic acid content increase, pH value the enzymolysis such as cellulose in the clover, has suppressed harmful fungus growth; In addition, can be monose with part soluble polysaccharide Substance Transformation, promote lactobacter growth, reach the purpose of fast reducing pH and raising ensiling quality.
The concrete technical scheme of the present invention is as follows:
A kind of microbial additive for alfalfa haylage is characterized in that: be comprised of the MRS fluid nutrient medium of the bacteria suspension of aspergillus oryzae (Aspergillus oryzae) and streptococcus lactis (Streptococcus lactis) mass percent by 50～70%:50 ~ 30%.
In the mentioned microorganism additive, contain at least 1.0 * 10 in the bacteria suspension of every milliliter of aspergillus oryzae
8The spore of individual aspergillus oryzae; At least contain 4.0 * 10 in every milliliter of MRS fluid nutrient medium
8The cfu streptococcus lactis.
In the mentioned microorganism additive, contain 5.0 * 10 in per 10 milliliters of microbe additives
8Individual ~ 7.0 * 10
8The spore of individual aspergillus oryzae contains 1.2 * 10 in per 10 milliliters of microbe additives
9~ 2.0 * 10
9The cfu streptococcus lactis.
The preparation method of above-mentioned microbial additive for alfalfa haylage may further comprise the steps:
(1) preparation of aspergillus oryzae bacteria suspension
A. the cultivation of aspergillus oryzae seed: aspergillus oryzae is placed in the corn synthetic medium activates, get seed liquor, activation temperature is 28 ℃, and incubation time is 4～5 days;
B. the solid culture of aspergillus oryzae: the weight ratio mixing of wheat bran and corn flour being pressed 85%:15%, being adjusted to moisture behind the mixing is 50wt%, get solid medium, the concentration of above-mentioned aspergillus oryzae seed liquor by 0.5wt% is joined in the solid medium, 28 ℃ of lower cultivations 24～48 hours of temperature, get the aspergillus oryzae spore;
C. aspergillus oryzae collecting cells: the above-mentioned solid medium that contains aspergillus oryzae of 10 grams is dissolved in 100 ml sterile waters, gets the aspergillus oryzae bacteria suspension;
(2) activation of streptococcus lactis: streptococcus lactis is placed in the MRS fluid nutrient medium activates, activation temperature is 37 ℃, soak time 36 hours;
(3) percentage by weight of the streptococcus lactis of aspergillus oryzae bacteria suspension and activation being pressed 50～70%:50～30% mixes, and gets microbe additive.
Among the above-mentioned preparation method, the component of described corn flour synthetic medium and content are: corn flour 5 grams, KH
2PO
40.3 gram, yeast extract 0.3 gram, glucose 1 gram, MgSO
47H
2O 1 gram, 1000 milliliters in water (being preferably distilled water), pH are about neutrality, sterilize 15 minutes for 121 ℃.
Among the above-mentioned preparation method, the component of described MRS fluid nutrient medium and content are: peptone 10 grams, beef extract 10 grams, yeast extract 5 grams, glucose 20 grams, KH
2PO
42 grams, calcium carbonate 10 grams, ammonium citrate 2 grams, 1 milliliter of Tween 80, agar 20 grams, MgSO
47H
2O 0.58 gram, MnSO44H
2O 0.25 gram, sodium acetate 5 grams, pH=6.4 sterilized 15 minutes for 121 ℃.
The application of above-mentioned microbial additive for alfalfa haylage is characterized in that, the per kilogram clover adds 10 ~ 15 milliliters of microbe additives during ensiling.
Microbe additive of the present invention in use should be cool extremely half-dried with clover forage, makes its water content between 55 ~ 60wt%, and silage effect can be better like this.
The present invention is wrapped up in the bag ensilage at the half-dried bale of clover and is tied the oxygen of wrapping up in the bag by adding the aspergillus oryzae eliminating, accelerate the initial fermenting speed of lactic acid bacteria, strengthen acid producing ability, effectively suppress the activity of saccharomycete and other undesirable microorganism, strengthened silage effect.Use this microbe additive than using separately lactic acid bacteria further to reduce pH, improved alfalfa silage quality, promoted the performance of milk cow, beef cattle, mutton sheep, pig production performance.Lactic acid bacteria commonly used when the used streptococcus lactis of the present invention is ensiling, used aspergillus oryzae (Aspergillus oryzae) is a kind of fungi commonly used in the food industry, be used in the production making soy sauce, the saccharifying agent of yellow rice wine, also be used for fermenting and producing cellulase, kojic acid etc.This two kinds of bacterial classifications all are simple and easy to, and therefore, this additive can be mass-produced using, and has practicality.
The specific embodiment
Below by specific embodiment the present invention is further set forth, should be understood that, following explanation only is exemplary, the present invention is not further set forth.Following content or concentration are mass percent if no special instructions.
Culture medium used among the following embodiment is as follows: corn flour synthetic medium prescription is: corn flour 5 grams, KH
2PO
40.3 gram, yeast extract 0.3 gram, glucose 1 gram, MgSO
47H
2O 1 gram, 1000 milliliters of distilled water were sterilized 15 minutes for 121 ℃.
MRS Liquid Culture based formulas is: peptone 10 grams, beef extract 10 grams, yeast extract 5 grams, glucose 20 grams, KH
2PO
42 grams, calcium carbonate 10 grams, ammonium citrate 2 grams, 1 milliliter of Tween 80, agar 20 grams, MgSO
47H
2O 0.58 gram, MnSO44H
2O 0.25 gram, sodium acetate 5 grams, pH=6.4 sterilized 15 minutes for 121 ℃.
Embodiment 1
1, the cultivation of aspergillus oryzae seed: aspergillus oryzae is placed in the corn synthetic medium activates, activation temperature is 28 ℃, incubation time 4 ~ 5 days.
2, the solid culture of aspergillus oryzae: the weight ratio mixing of wheat bran and corn flour being pressed 85%:15%, being adjusted to moisture behind the mixing is 50%, get solid medium, above-mentioned aspergillus oryzae seed liquor is added in the solid medium by 0.5% concentration, under 28 ℃ of temperature, cultivated 24 ~ 48 hours, produce green aspergillus oryzae spore, its spore germination rate ﹥ 99%.After the cultivation, contain at least spore 10 in every gram solid medium
9Individual.
3, aspergillus oryzae collecting cells: above-mentioned 10 gram solid mediums are dissolved in 100 ml sterile waters, make in every milliliter of bacteria suspension and contain at least 10
8Individual spore.
4, the activation of streptococcus lactis: streptococcus lactis is placed on the activation of MRS fluid nutrient medium, and activation temperature is 37 ℃, and soak time 36 hours when activation is finished, contains streptococcus lactis 4 * 10 in every milliliters of liquid culture medium at least
8Cfu.
5, the microbe additive of alfalfa ensilage is made: the streptococcus lactis that activates in aspergillus oryzae bacteria suspension and the MRS fluid nutrient medium is mixed in the ratio of 50%:50%, form the microbe additive of alfalfa ensilage, wherein per 10 milliliters of these microbe additives contain aspergillus oryzae 5.0 * 10
8Individual, lactic acid hammer 2.0 * 10
9Cfu, per kilogram adds 10 milliliters of microbe additives during ensiling.
After using embodiment 1 made microbe additive that clover is carried out ensiling, contained protein, lactic acid bacteria, lactic acid and pH effect see Table 1 in the clover.
As seen from Table 1, with the clover that this microbe additive is processed, through 40 ~ 60 days ensiling, the indices of ensiling all reached good effect.The pH value is processed low by 11.3% than streptococcus lactis, protein and lactic acid content are processed than streptococcus lactis respectively and improved 7.1% and 9.0%, and the quantity of lactic acid bacteria has increased by 10 times more than, and difference has all reached the level of signifiance.
Use this microbe additive Alfalfa Silage feeding cow, 30 days 50 cow head feeding experiments show that every of average daily ingestion amount improves 3%, and the output of milk improves 6.8%.
Embodiment 2
1, the cultivation of aspergillus oryzae seed: aspergillus oryzae is placed in the corn synthetic medium activates, activation temperature is 28 ℃, incubation time 4 ~ 5 days.
2, the solid culture of aspergillus oryzae: the weight ratio mixing of wheat bran and corn flour being pressed 85%:15%, being adjusted to moisture behind the mixing is 50%, get solid medium, above-mentioned aspergillus oryzae seed liquor is added in the solid medium by 0.5% concentration, cultivated 24 ~ 48 hours, produce green spores, its spore germination rate ﹥ 99%.After the cultivation, contain at least spore 10 in every gram solid medium
9Individual.
3, aspergillus oryzae collecting cells: 10 gram solid mediums are dissolved in 100 ml sterile waters, make in every milliliter of bacteria suspension and contain at least 10
8Individual spore.
4, the activation of streptococcus lactis: streptococcus lactis is placed on the activation of MRS fluid nutrient medium, and activation temperature is 37 ℃, and soak time 36 hours when activation is finished, contains streptococcus lactis 4.0 * 10 in every milliliters of liquid culture medium at least
8Cfu.
5, the microbe additive of alfalfa ensilage is made: the streptococcus lactis that activates in aspergillus oryzae bacteria suspension and the MRS fluid nutrient medium is mixed in the ratio of 60%:40%, form the microbe additive of alfalfa ensilage, wherein per 10 milliliters of these microbe additives contain aspergillus oryzae 6.0 * 10
8Individual, lactic acid hammer 1.6 * 10
9Cfu, per kilogram adds 10 milliliters of microbe additives during ensiling.
After using embodiment 2 made microbe additives that clover is carried out ensiling, contained protein, lactic acid bacteria, lactic acid and pH effect see Table 2 in the clover.
As seen from Table 2, with the clover of this additive treating, through 40 ~ 60 days ensiling, the indices of ensiling all reached good effect.The pH value is processed low by 10.3% than streptococcus lactis, protein and lactic acid content are processed than streptococcus lactis respectively and improved 6.8% and 10.9%, and the quantity of lactic acid bacteria has increased by 10 times more than, and difference has all reached the level of signifiance.
Use this additive Alfalfa Silage beef cattle of feeding, 40 beef cattle feedings tests in 60 days show that every of average daily ingestion amount improves 2.6%, and daily gain improves 5.7%.
Embodiment 3
1, the cultivation of aspergillus oryzae seed: aspergillus oryzae is placed in the corn synthetic medium activates, activation temperature is 28 ℃, incubation time 4～5 days.
2, the solid culture of aspergillus oryzae: the weight ratio mixing of wheat bran and corn flour being pressed 85%:15%, being adjusted to moisture behind the mixing is 50%, get solid medium, above-mentioned aspergillus oryzae seed liquor is added in the solid medium by 0.5% concentration, cultivated 24 ~ 48 hours, produce green spores, its spore germination rate ﹥ 99%.After the cultivation, contain at least spore 10 in every gram solid medium
9Individual.
3, aspergillus oryzae collecting cells: 10 gram solid mediums are dissolved in 100 ml sterile waters, make in every milliliter of bacteria suspension and contain at least 10
8Individual spore.
4, the activation of streptococcus lactis: streptococcus lactis is placed on the activation of MRS fluid nutrient medium, and activation temperature is 37 ℃, and soak time 36 hours when activation is finished, contains streptococcus lactis 4 * 10 in every milliliters of liquid culture medium at least
8Cfu.
5, the microbe additive of alfalfa ensilage is made: the streptococcus lactis that activates in aspergillus oryzae bacteria suspension and the MRS fluid nutrient medium is mixed in the ratio of 70%:30%, form the microbe additive of alfalfa ensilage, wherein per 10 milliliters of these microbe additives contain aspergillus oryzae 7 * 10
8Individual, lactic acid hammer 1.2 * 10
9Cfu, per kilogram adds 15 milliliters of microbe additives during ensiling.
After using embodiment 3 made microbe additives that clover is carried out ensiling, contained protein, lactic acid bacteria, lactic acid and pH effect see Table 3 in the clover.
As seen from Table 3, with the clover of this additive treating, through 40 ~ 60 days ensiling, the indices of ensiling all reached good effect.The pH value is processed low by 9.27% than streptococcus lactis, protein and lactic acid content and lactic acid bacterium number are processed than streptococcus lactis respectively and improved 6.6% and 8.85%, and the quantity of lactic acid bacteria has increased by 10 times more than, and difference has all reached the level of signifiance.
Use this additive Alfalfa Silage mutton sheep of feeding, 60 days 100 mutton sheep feeding experiments show that every of average daily ingestion amount improves 2.6%, and daily gain improves 5.8%.
Claims (3)
1. microbial additive for alfalfa haylage is characterized in that: be comprised of the MRS fluid nutrient medium of the bacteria suspension of aspergillus oryzae (Aspergillus oryzae) and streptococcus lactis (Streptococcus lactis) mass percent by 50～70%:50 ~ 30%; At least contain 1.0 * 10 in the bacteria suspension of every milliliter of aspergillus oryzae
8The spore of individual aspergillus oryzae; At least contain 4.0 * 10 in every milliliter of MRS fluid nutrient medium
8The cfu streptococcus lactis contains 5.0 * 10 in per 10 milliliters of microbe additives
8Individual ~ 7.0 * 10
8The spore of individual aspergillus oryzae contains 1.2 * 10
9~ 2.0 * 10
9The cfu streptococcus lactis.
2. the preparation method of a microbial additive for alfalfa haylage claimed in claim 1 is characterized in that may further comprise the steps:
(1) preparation of aspergillus oryzae bacteria suspension
A. the cultivation of aspergillus oryzae seed: aspergillus oryzae is placed in the corn synthetic medium activates, get seed liquor, activation temperature is 28 ℃, and incubation time is 4～5 days;
B. the solid culture of aspergillus oryzae: the weight ratio mixing of wheat bran and corn flour being pressed 85%:15%, being adjusted to moisture behind the mixing is 50wt%, get solid medium, the concentration of above-mentioned aspergillus oryzae seed liquor by 0.5wt% is joined in the solid medium, 28 ℃ of lower cultivations 24～48 hours of temperature, get the aspergillus oryzae spore;
C. aspergillus oryzae collecting cells: the above-mentioned solid medium that contains aspergillus oryzae of 10 grams is dissolved in 100 ml sterile waters, gets the aspergillus oryzae bacteria suspension;
(2) activation of streptococcus lactis: streptococcus lactis is placed in the MRS fluid nutrient medium activates, activation temperature is 37 ℃, soak time 36 hours;
(3) percentage by weight of the streptococcus lactis of aspergillus oryzae bacteria suspension and activation being pressed 50～70%:50～30% mixes, and gets microbe additive;
The component of described corn synthetic medium and content are: corn flour 5 grams, KH
2PO
40.3 gram, yeast extract 0.3 gram, glucose 1 gram, MgSO
47H
2O 1 gram, 1000 milliliters in water was sterilized 15 minutes for 121 ℃.
3. the application of a microbial additive for alfalfa haylage claimed in claim 1 is characterized in that: the per kilogram clover adds 10 ~ 15 milliliters of microbe additives during ensiling, and during ensiling, the clover water content is 55 ~ 60wt%.
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|---|---|---|---|
| CN 201010224736 CN101869230B (en) | 2010-07-13 | 2010-07-13 | Microbial additive for alfalfa haylage, preparation method and application thereof |
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| CN101869230B true CN101869230B (en) | 2013-01-16 |
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Families Citing this family (7)
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| CN102860412A (en) * | 2011-07-06 | 2013-01-09 | 吉林大学 | Co-production method for distilled spirit and livestock total nutrient feed through alfalfa fermentation |
| CN102318729B (en) * | 2011-07-29 | 2014-02-26 | 林善斌 | Method and equipment for preparing low-moisture silage |
| CN102511699A (en) * | 2011-12-21 | 2012-06-27 | 金华职业技术学院 | Alfalfa silaging method |
| CN107027977A (en) * | 2017-04-25 | 2017-08-11 | 六安市绿丰牧业科技有限公司 | The preparation method that a kind of sheep eats ensilage |
| CN108740293A (en) * | 2018-05-21 | 2018-11-06 | 何怡江 | The method that microbial fermentation prepares Alfalfa feed |
| CN108813152A (en) * | 2018-06-29 | 2018-11-16 | 内蒙古农业大学 | A kind of caragana microphylla base solid state fermentation feed and preparation method thereof |
| CN112753867B (en) * | 2020-12-30 | 2023-02-28 | 广西壮族自治区畜牧研究所 | Feed for improving functions of cattle liver and application thereof |
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| CN1757714A (en) * | 2005-08-11 | 2006-04-12 | 上海交通大学 | Composite bacterial additive of alfalfa ensilage |
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