CN1718076A - Method for preparing microoganism additives for ensiling lucerne - Google Patents
Method for preparing microoganism additives for ensiling lucerne Download PDFInfo
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- CN1718076A CN1718076A CNA2005100286902A CN200510028690A CN1718076A CN 1718076 A CN1718076 A CN 1718076A CN A2005100286902 A CNA2005100286902 A CN A2005100286902A CN 200510028690 A CN200510028690 A CN 200510028690A CN 1718076 A CN1718076 A CN 1718076A
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Abstract
A microbial additive for the alfalfa ensilage is prepared through choosing fresh alfalfa, preparing fermented green juice, separating and purifying streptococcus lactis, activating, and proportionally mixing said fermented green juice with the activated streptococcus lactis in the liquid MRS culture medium. Its advantage is high quality of alfalfa ensilage.
Description
Technical field
That the present invention relates to is a kind of preparation method of biological technical field, specifically is a kind of preparation method of microoganism additives for ensiling lucerne.
Background technology
Alfalfa (Medicago sativa L. is hereinafter to be referred as clover) is a kind of perennial leguminous forage, and nutritive value is abundant, is the forage grass of plant-eating animal high-quality, and the good reputation of " king of herbage " is arranged.For a long time, the preservation of alfalfa is mainly based on hay curing, and the disadvantage of this method is that the nutriment loss is serious.In recent years, Canada, Japan and other countries began to utilize the method for ensiling to preserve clover, had solved the serious problem of nutriment loss in the clover preservation process.Because the clover soluble sugar content is low, the buffer capacity height belongs to the leguminous forage of difficult ensiling, and directly the ensiling effect is bad, need use additive in the silage fermentation process.The commonplace additive types of external use at present has the mixture of formic acid, lactobacillus preparation, enzyme preparation and lactic acid bacteria and enzyme preparation etc.
Find through literature search prior art, (1997) such as Mitsuaki Ohshima propose the preparation method of fermented green juice (PreviouslyFermented Juice is abbreviated as PFJ) and the effect in alfalfa ensilage thereof first at Animal FeedScience Technology the 66th phase of magazine (129-137 page or leaf).The main manufacturing process of this method is as follows: fresh clover → shred → add adds in water → dipping → pulverizing (making beating) → filtration → filtrate that glucose → PFJ ferments → make.Because fermented green juice contains the multiple lactic acid bacteria culturers that derives from clover, the effect of obvious reduction ensilage pH value is arranged in alfalfa ensilage, and, fermented green juice is made simple, draw materials conveniently, have no side effect behind the edible ensilage of domestic animal, harmless in the ensiling manufacturing process, therefore, begin gradually to be applied aborning.As the lactic acid fermented startup factor, fermented green juice can promote the lactic acid bacteria population to enlarge in ensilage, suppresses the growth of other harmful bacterium, improves the alfalfa ensilage quality.Many studies have shown that its effect is more effective than the single lactic acid bacteria of inoculation, can significantly reduce the pH value of ensiling clover with fermented green juice ensiling clover.Yet, in the fermented green juice except that containing various lactobacillus, also contain multiple harmful bacterial classification, these bacterial classifications equally also have the effect that starts the factor, in the alfalfa ensilage process, induce harmful bacterial classification group to enlarge, thereby reduced or slowed down the effect that fermented green juice promotes the lactic acid bacteria expanding propagation better, make the ensiling clover can not sharply reduce the pH value of ensilage early stage in a short time in preparation yeast phase and acidifying, this be fermented green juice as the greatest drawback that additives for ensiling exists in alfalfa ensilage, have influence on ensiling clover high-quality fermentation and form high-quality ensilage.In further retrieval, yet there are no so far relevant from clover separating lactic acid streptococcus (Streptococcus lactis) do not see yet fermented green juice and streptococcus lactis mixed report as addictive for alfalfa silage as the report of additives for ensiling.
Summary of the invention
The present invention is directed to the deficiency and the defective of fermented green juice ensiling in the background technology, a kind of preparation method of microoganism additives for ensiling lucerne is provided, make its gained microoganism additives for ensiling lucerne, obvious reduction ensiling preparation yeast phase and acidifying clover pH in early stage value are arranged, improve the effect of ensiling quality.
The present invention is achieved by the following technical solutions, microoganism additives for ensiling lucerne of the present invention is made up of by the percentage by weight of 50-70%:30-50% fermented green juice (PFJ) and the MRS fluid nutrient medium of preserving streptococcus lactis, contains streptococcus lactis 2 * 10 at least in wherein every ml MRS fluid nutrient medium
7Cfu, method step is as follows:
(1) selects fresh clover material;
(2) making of fermented green juice;
(3) streptococcus lactis separation, purge process;
(4) streptococcus lactis activation;
(5) fermented green juice is mixed in proportion with the streptococcus lactis of activation, form microoganism additives for ensiling lucerne.
Described step (1) is specially: get fresh alfalfa plants, remove each about 10cm of base portion and top, remove major branch and thicker branch simultaneously, remaining sprout and leaf are kept, as the making material of fermented green juice;
Described step (2), be specially: fresh clover material is chopped into 3-5cm, add distilled water in clover (g) and 1: 2.5 ratio of water (ml), fully mix, with organizing pulverizer to smash to pieces at a high speed, left standstill 1 hour, double gauze filters, and adds the glucose or the suitable syrup of glucose amount, abundant stirring by the ratio of clover raw material: glucose=10g: 1g in filtrate, place 30 ℃ environment to ferment 48 hours, make fermented green juice;
Described step (3), be specially: with 60～100 times of fermented green juice dilutions, place the separating solids culture medium, 48h are cultivated down for 35 ~ 40 ℃ in medium pH=5.8～6.4, select that molten calcium circle is big, the bacterium colony of culture medium flavescence, be placed on MRS solid medium purifying, obtain streptococcus lactis (ST), the streptococcus lactis behind the purifying is preserved with the test tube that the MRS solid medium is housed, and the product acidity of this streptococcus lactis in 48 hours can see Table 1.
Described separating solids culture medium (%) prescription is: beef extract 1g, and peptone 1g, yeast extract 1g, glucose 1g, tomato juice 20%, soil temperature 80 0.05%, calcium carbonate 2g, bromocresol green 0.01%, agar 1.5g, pH 5.8～6.4.
Described MRS solid medium (%) prescription is: peptone 1g, beef extract 1g, yeast extract 0.5g, K
2HPO
40.2g, dibasic ammonium citrate 0.2g, sodium acetate 0.5g, glucose 2g, soil temperature 80 0.1%, MgSO
4.7H
2O 0.058g, MnSO
4.4H
2O 0.025g, agar 1.5g, pH=6.2,121 ℃ of sterilization 15min.
The pH value of table 1 ST strain cultured solution and lactic acid content change (mmol/L)
| 0h | 6h | 12h | 24h | 48h | |
| PH | 6.09±0.02 | 4.87±0.04 | 4.34±0.02 | 3.99±0.01 | 4.02±0.01 |
| Lactic acid content | 0.50±0.03 | 3.30±0.03 | 17.74±0.43 | 27.91±1.30 | 36.35±1.29 |
Described step (4) is specially: the streptococcus lactis that separates is placed in the MRS fluid nutrient medium activates, and 35～37 ℃ of activation temperatures, soak time 24～36 hours when activation is finished, contains streptococcus lactis 2 * 10 in every ml fluid nutrient medium at least
7Cfu.
Described MRS fluid nutrient medium (%) prescription is: peptone 1g, beef extract 1g, yeast extract 0.5g, K
2HPO
40.2g, dibasic ammonium citrate 0.2g, sodium acetate 0.5g, glucose 2g, soil temperature 80 0.1%, MgSO
4.7H
2O0.058g, MnSO
4.4H
2O 0.025g, pH=6.2,121 ℃ of sterilization 15min.
Described step (5), be specially: fermented green juice is mixed in the ratio of 50-70%: 30-50% with streptococcus lactis alive in the MRS fluid nutrient medium, form microoganism additives for ensiling lucerne, wherein every this additive of 10ml contains PFJ5-7ml, streptococcus lactis 10
7～10
8Cuf, the per kilogram clover adds the 10-15ml additive during ensiling.
Characteristics of the present invention are: (1) streptococcus lactis is by separating in the clover; (2) two additives that have complementary functions of this streptococcus lactis and PFJ are mixed in proportion, form this additive, it is different from external streptococcus lactis (by obtaining in the non-clover material) or the PFJ method as addictive for alfalfa silage of using separately.Fermented green juice is the lactic acid fermented startup factor, and streptococcus lactis is the lactic acid bacteria culturers that the preparation yeast phase plays a major role, it is the fastest lactic acid bacteria culturers of preparation yeast phase breeding, two kinds of additives comprehensive and complementary, strengthened fermented green juice and promoted the effect that the ensiling clover is fermented, made this additive use the effect that better reduction pH and raising ensiling quality are arranged separately than fermented green juice and these two kinds of additives of streptococcus lactis.With the clover of this additive treating, through ensiling in 30 days, ensilage pH value reduced by 10.4～12.7% than handling with fermented green juice.Gained microoganism additives for ensiling lucerne of the present invention has obvious reduction ensiling preparation yeast phase and acidifying clover pH in early stage value, improves the effect of ensiling quality.
The specific embodiment
Provide following examples in conjunction with the inventive method step:
Embodiment one
1, selects fresh clover material: get fresh alfalfa plants, remove each about 10cm of base portion and top, remove major branch and thicker branch simultaneously, remaining sprout and leaf are kept, as the making material of fermented green juice;
2, fermented green juice is made: fresh clover material is chopped into 3-5cm, add distilled water in clover (g) and 1: 2.5 ratio of water (ml), fully mix, with organizing pulverizer to smash to pieces at a high speed, left standstill 1 hour, double gauze filters, the syrup that in filtrate, adds glucose or suitable glucose amount in the ratio of clover raw material: glucose=10g: 1g, fully stir, place 30 ℃ of fermentations 48 hours, make fermented green juice;
3, streptococcus lactis separation, purifying: with 100 times of fermented green juice dilutions, place the separating solids culture medium, medium pH=5.8, cultivate 48h down for 35 ℃, select that molten calcium circle is big, the bacterium colony of culture medium flavescence, be placed on MRS solid medium purifying, obtain streptococcus lactis (ST), the streptococcus lactis behind the purifying is preserved with the test tube that the MRS solid medium is housed.
Described separating solids culture medium (%) prescription is: beef extract 1g, peptone 1g, yeast extract 1g, glucose 1g, tomato juice 20%, soil temperature 80 0.05%, calcium carbonate 2g, bromocresol green 0.01%, agar 1.5g, pH=5.8.
Described MRS solid medium (%) prescription is: peptone 1g, beef extract 1g, yeast extract 0.5g, K
2HPO
40.2g, dibasic ammonium citrate 0.2g, sodium acetate 0.5g, glucose 2g, soil temperature 80 0.1%, MgSO
4.7H
2O 0.058g, MnSO
4.4H
2O 0.025g, agar 1.5g, pH=6.2,121 ℃ of sterilization 15min.
4, streptococcus lactis activation: the streptococcus lactis of separator well is placed in the MRS fluid nutrient medium activates, 35 ℃ of activation temperatures, soak time 24 hours when activation is finished, contains streptococcus lactis 2 * 10 in every ml fluid nutrient medium at least
7Cfu.
Described MRS fluid nutrient medium (%) prescription is: peptone 1g, beef extract 1g, yeast extract 0.5g, K
2HPO
40.2g, dibasic ammonium citrate 0.2g, sodium acetate 0.5g, glucose 2g, soil temperature 80 0.1%, MgSO
4.7H
2O0.058g, MnSO
4.4H
2O 0.025g, pH=6.2,121 ℃ of sterilization 15min.
5, the preparation of microoganism additives for ensiling lucerne: fermented green juice is mixed in 50%: 50% ratio with streptococcus lactis alive in the MRS fluid nutrient medium, form microoganism additives for ensiling lucerne, wherein every this additive of 10ml contains PFJ 5ml, streptococcus lactis 10
8Cuf, the per kilogram clover adds the 10ml additive during ensiling.
The alfalfa ensilage effect of table 2 embodiment one
| Handle | Sensible quality | pH | a＝0.05 | Protein | a＝0.05 |
| Additive-free | Inferior | 5.10±0.12 | a | 17.25±0.51％ | c |
| PFJ | Medium | 4.87±0.16 | b | 20.31±0.12％ | b |
| This additive | Good | 4.32±0.18 | c | 22.07±0.19％ | a |
As seen from Table 2, with the clover of this additive treating, through ensiling in 30 days, the ensilage sensory evaluation reached good, and the sensory evaluation of PFJ is medium; Than PFJ processing low 12.7% and raising 8.7%, difference all reaches the level of signifiance respectively for pH value and protein content.
Embodiment two
1, selects fresh clover material: get fresh alfalfa plants, remove each about 10cm of base portion and top, remove major branch and thicker branch simultaneously, remaining sprout and leaf are kept, as the making material of fermented green juice;
2, fermented green juice is made: fresh clover material is chopped into 3-5cm, add distilled water in clover (g) and 1: 2.5 ratio of water (ml), fully mix, with organizing pulverizer to smash to pieces at a high speed, left standstill 1 hour, double gauze filters, the syrup that in filtrate, adds glucose or suitable glucose amount in the ratio of clover raw material: glucose=10g: 1g, fully stir, place 30 ℃ of fermentations 48 hours, make fermented green juice;
3, streptococcus lactis separation, purifying: with 80 times of fermented green juice dilutions, place the separating solids culture medium, medium pH=6.1, cultivate 48h down for 37 ℃, select that molten calcium circle is big, the bacterium colony of culture medium flavescence, be placed on MRS solid medium purifying, obtain streptococcus lactis (ST), the streptococcus lactis behind the purifying is preserved with the test tube that the MRS solid medium is housed;
Described separating solids culture medium (%) prescription is: beef extract 1g, peptone 1g, yeast extract 1g, glucose 1g, tomato juice 20%, soil temperature 80 0.05%, calcium carbonate 2g, bromocresol green 0.01%, agar 1.5g, pH=6.1.
Described MRS solid medium (%) prescription is identical with embodiment one.
4, streptococcus lactis activation: the streptococcus lactis that separates is placed in the MRS fluid nutrient medium activates, 37 ℃ of activation temperatures, soak time 30 hours when activation is finished, contains streptococcus lactis 2 * 10 in every ml fluid nutrient medium at least
7Cfu.
Described MRS fluid nutrient medium (%) prescription is identical with embodiment one.
5, the preparation of microoganism additives for ensiling lucerne: fermented green juice is mixed in 60%: 40% ratio with streptococcus lactis alive in the MRS fluid nutrient medium, form microoganism additives for ensiling lucerne, wherein every this additive of 10ml contains PFJ6ml, streptococcus lactis 8 * 10
7Cuf, the per kilogram clover adds the 12ml additive during ensiling.
The alfalfa ensilage effect of table 3 embodiment two
| Handle | Sensible quality | pH | α＝0.05 | Protein | α＝0.05 |
| Additive-free | Inferior | 5.10±0.12 | a | 17.25±0.51％ | c |
| PFJ | Medium | 4.87±0.16 | b | 20.31±0.12％ | b |
| This additive | Good | 4.38±0.21 | c | 22.17±0.24％ | a |
As seen from Table 3, with the clover of this additive treating, through ensiling in 30 days, the ensilage sensory evaluation reached good, and the sensory evaluation of PFJ is medium; Than PFJ processing low 11.9% and raising 9.2%, difference all reaches the level of signifiance respectively for pH value and protein content.
Embodiment three
1, selects fresh clover material: get fresh alfalfa plants, remove each about 10cm of base portion and top, remove major branch and thicker branch simultaneously, remaining sprout and leaf are kept, as the making material of fermented green juice;
2, fermented green juice is made: fresh clover material is chopped into 3-5cm, add distilled water in clover (g) and 1: 2.5 ratio of water (ml), fully mix, with organizing pulverizer to smash to pieces at a high speed, left standstill 1 hour, double gauze filters, the syrup that in filtrate, adds glucose or suitable glucose amount in the ratio of clover raw material: glucose=10g: 1g, fully stir, place 30 ℃ of fermentations 48 hours, make fermented green juice;
3, streptococcus lactis separation, purifying: with 60 times of fermented green juice dilutions, place solid medium, medium pH=6.4, cultivate 48h down for 40 ℃, select that molten calcium circle is big, the bacterium colony of culture medium flavescence, be placed on MRS solid medium purifying, obtain streptococcus lactis (ST), the streptococcus lactis behind the purifying is preserved with the test tube that the MRS solid medium is housed;
Described solid medium (%) prescription is: beef extract 1g, peptone 1g, yeast extract 1g, glucose 1g, tomato juice 20%, soil temperature 80 0.05%, calcium carbonate 2g, bromocresol green 0.01%, agar 1.5g, pH=6.4.
Described MRS solid medium (%) prescription is identical with embodiment one.
4, streptococcus lactis activation: the streptococcus lactis of separator well is placed in the MRS fluid nutrient medium activates, 37 ℃ of activation temperatures, soak time 36 hours when activation is finished, contains streptococcus lactis 2 * 10 in every ml fluid nutrient medium at least
7Cfu.
Described MRS fluid nutrient medium (%) prescription is identical with embodiment one.
5, the preparation of microoganism additives for ensiling lucerne: fermented green juice is mixed in 70%: 30% ratio with streptococcus lactis alive in the MRS fluid nutrient medium, form microoganism additives for ensiling lucerne, wherein every this additive of 10ml contains PFJ 7ml, streptococcus lactis 6 * 10
7Cuf, the per kilogram clover adds the 15ml additive during ensiling.
The alfalfa ensilage effect of table 4 embodiment three
| Handle | Sensible quality | pH | α＝0.05 | Protein | α＝0.05 |
| Additive-free | Inferior | 5.10±0.12 | a | 17.25±0.51％ | c |
| PFJ | Medium | 4.87±0.02 | b | 20.31±0.12％ | b |
| This additive | Good | 4.41±0.18 | c | 22.01±0.19％ | a |
As seen from Table 4, with the clover of this additive treating, through ensiling in 30 days, the ensilage sensory evaluation reached good, and the sensory evaluation of PFJ is medium; Than PFJ processing low 10.4% and raising 8.4%, difference all reaches the level of signifiance respectively for pH value and protein content.
The preparation method of above-mentioned three embodiment and the technological process of production can be sketched and be:
Fresh clover → selected → chopping → dipping → pulverizing (making beating) → leave standstill → filter → the add glucose → PFJ → dilution of fermenting → make → streptococcus lactis cultivation → purifying → acquisition streptococcus lactis → activation → PFJ+ streptococcus lactis mixing → microoganism additives for ensiling lucerne.
Making difference between table 5 embodiment
| Embodiment | The PFJ extension rate | Medium pH | Cultivation temperature ℃ | Soak time h | Activation temperature ℃ |
| 1 | 100 | 5.8 | 35 | 24 | 35 |
| 2 | 80 | 6.1 | 38 | 30 | 37 |
| 3 | 60 | 6.4 | 40 | 36 | 37 |
Claims (10)
1, a kind of preparation method of microoganism additives for ensiling lucerne, it is characterized in that, described microoganism additives for ensiling lucerne is made up of by the percentage by weight of 50-70%:30-50% fermented green juice and the MRS fluid nutrient medium of preserving streptococcus lactis, and preparation process is as follows:
(1) selects fresh clover material;
(2) making of fermented green juice:
(3) streptococcus lactis separation, purge process;
(4) streptococcus lactis activation;
(5) fermented green juice is mixed in proportion with the streptococcus lactis of activation, form microoganism additives for ensiling lucerne.
2, the preparation method of microoganism additives for ensiling lucerne according to claim 1 is characterized in that, described step (1), be specially: get fresh alfalfa plants, remove each 10cm of base portion and top, remove major branch and thicker branch simultaneously, remaining sprout and leaf are kept.
3, the preparation method of microoganism additives for ensiling lucerne according to claim 1 is characterized in that, described step (2) is specially: the fresh alfalfa material that (a) will choose is chopped into 3-5cm; (b) in clover g and water ml1: 2.5 ratio adds distilled water, fully mix, with organizing pulverizer that mixed clover is smashed to pieces at a high speed, left standstill 1 hour, filter through double gauze: the syrup that (c) in filtrate, adds glucose or suitable glucose amount in the ratio of clover raw material: glucose=10g: 1g, fully stir, place 30 ℃ environment to ferment 48 hours, make fermented green juice.
4, the preparation method of microoganism additives for ensiling lucerne according to claim 1 is characterized in that, described step (3), be specially: (a) with 60～100 times of fermented green juice dilutions, place the separating solids culture medium, medium pH=5.8～6.4 are cultivated 48h down for 35 ~ 40 ℃; (b) select that molten calcium circle is big, the bacterium colony of culture medium flavescence, be placed on MRS solid medium purifying, obtain streptococcus lactis.
5, the preparation method of microoganism additives for ensiling lucerne according to claim 4, it is characterized in that described separating solids culture medium prescription is: beef extract 1g, peptone 1g, yeast extract 1g, glucose 1g, tomato juice 20%, soil temperature 800.05%, calcium carbonate 2g, bromocresol green 0.01%, agar 1.5g, pH5.8～6.4.
6, the preparation method of microoganism additives for ensiling lucerne according to claim 4 is characterized in that, described MRS solid culture based formulas is: peptone 1g, beef extract 1g, yeast extract 0.5g, K
2HPO
40.2g, dibasic ammonium citrate 0.2g, sodium acetate 0.5g, glucose 2g, soil temperature 80 0.1%, MgSO
4.7H
2O 0.058g, MnSO
4.4H
2O 0.025g, agar 1.5g, pH=6.2,121 ℃ of sterilization 15min.
7, the preparation method of microoganism additives for ensiling lucerne according to claim 1, it is characterized in that, described step (4), be specially: the streptococcus lactis of separator well is placed in the MRS fluid nutrient medium activates, 35～37 ℃ of activation temperatures, soak time 24～36 hours when activation is finished, contains streptococcus lactis 2 * 10 in every ml MRS fluid nutrient medium at least
7Cfu.
8, the preparation method of microoganism additives for ensiling lucerne according to claim 7 is characterized in that, described MRS Liquid Culture based formulas is: peptone 1g, beef extract 1g, yeast extract 0.5g, K
2HPO
40.2g, dibasic ammonium citrate 0.2g, sodium acetate 0.5g, glucose 2g, soil temperature 80 0.1%, MgSO
4.7H
2O 0.058g, MnSO
4.4H
2O 0.025g, pH=6.2,121 ℃ of sterilization 15min.
9, the preparation method of microoganism additives for ensiling lucerne according to claim 1, it is characterized in that, described step (5), be specially: fermented green juice is mixed in the ratio of 50-70%: 30-50% with streptococcus lactis alive in the MRS fluid nutrient medium, form microoganism additives for ensiling lucerne.
10, the preparation method of microoganism additives for ensiling lucerne according to claim 9 is characterized in that, wherein every 10ml additive contains PFJ 5-7ml, streptococcus lactis 10
7～10
8Cuf, the per kilogram clover adds the 10-15ml additive during ensiling.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101869230A (en) * | 2010-07-13 | 2010-10-27 | 山东省农业科学院畜牧兽医研究所 | Microbial additive for alfalfa haylage, preparation method and application thereof |
| 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 |
| CN103053808A (en) * | 2013-01-25 | 2013-04-24 | 中国农业大学 | Silage additive and preparation method and application thereof |
| CN104193548A (en) * | 2014-08-11 | 2014-12-10 | 上海交通大学 | Preparation method of biological additive for improving effect of alfalfa green manure |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE68918471D1 (en) * | 1988-02-18 | 1994-11-03 | Calpis Food Ind Co Ltd | Lactic acid bacteria starter for the production of feed and an agent containing these viable bacteria. |
| RU1799395C (en) * | 1990-09-10 | 1993-02-28 | Институт Микробиологии И Вирусологии Ан@ Казсср | Strain of bacterium acidocaldarius used for straw ensilage |
| US6455063B1 (en) * | 1998-04-17 | 2002-09-24 | The Board Of Regents For Oklahoma State University | Propionibacterium P-63 for use in direct fed microbials for animal feeds |
| CN1297654C (en) * | 2003-03-14 | 2007-01-31 | 新疆威仕达生物工程股份有限公司 | Microbial ensiling strain and composite fungus, method for producing silage |
-
2005
- 2005-08-11 CN CNB2005100286902A patent/CN1324980C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101869230A (en) * | 2010-07-13 | 2010-10-27 | 山东省农业科学院畜牧兽医研究所 | Microbial additive for alfalfa haylage, preparation method and application thereof |
| CN101869230B (en) * | 2010-07-13 | 2013-01-16 | 山东省农业科学院畜牧兽医研究所 | Microbial additive for alfalfa haylage, preparation method and application thereof |
| 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 |
| CN103053808A (en) * | 2013-01-25 | 2013-04-24 | 中国农业大学 | Silage additive and preparation method and application thereof |
| CN103053808B (en) * | 2013-01-25 | 2014-01-08 | 中国农业大学 | Silage additive and preparation method and application thereof |
| CN104193548A (en) * | 2014-08-11 | 2014-12-10 | 上海交通大学 | Preparation method of biological additive for improving effect of alfalfa green manure |
| CN104193548B (en) * | 2014-08-11 | 2016-05-04 | 上海交通大学 | The bio-additive preparation method of a kind of alfalfa green manure synergy |
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| Publication number | Publication date |
|---|---|
| CN1324980C (en) | 2007-07-11 |
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