CN1757714A - Composite bacterial additive of alfalfa ensilage - Google Patents
Composite bacterial additive of alfalfa ensilage Download PDFInfo
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- CN1757714A CN1757714A CNA2005100286955A CN200510028695A CN1757714A CN 1757714 A CN1757714 A CN 1757714A CN A2005100286955 A CNA2005100286955 A CN A2005100286955A CN 200510028695 A CN200510028695 A CN 200510028695A CN 1757714 A CN1757714 A CN 1757714A
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- streptococcus
- additive
- alfalfa
- acidi lactici
- green juice
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Abstract
A composite microbial additive for preparing the ensilage of alfalfa is composed of the fermented liquid of green juice and the liquid culture medium MRS containing lactosteptococcus and faecal steptococcus. Its preparing process includes such steps as preparing the fermented liquid of green juice, separating lactosteptococcus, activating lactosteptococcus and faecal steptococcus, and proportional mixing. It can decrease the pH value of ensilage and increase its quality.
Description
Technical field
What the present invention relates to is a kind of additive of biological technical field, specifically is a kind of composite bacterial additive of alfalfa ensilage.
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 phytophagous animal fine forage grass, 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 nutritive substance 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 nutritive substance 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, lactic acid bacteria formulation, zymin and milk-acid bacteria and zymin etc.The research of China aspect alfalfa ensilage still is in the starting stage, the research of addictive for alfalfa silage is still belonged to blank, but China's alfalfa ensilage market potential is huge, and the market outlook of additives for ensiling are fine.
Find through literature search prior art, (1997) such as Mitsuaki Ohshima propose the making method of fermented green juice (Previously Fermented Juice is abbreviated as PFJ) and the effect in alfalfa ensilage thereof first at AnimalFeed Science Technology the 66th phase of magazine (129-137 page or leaf).The main making processes 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 making processes, therefore, begin gradually to be applied aborning.As the lactic acid fermented startup factor, fermented green juice can promote the milk-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 milk-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 deleterious 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 milk-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, Shang Weijian mixes report as addictive for alfalfa silage with fermented green juice with streptococcus acidi lactici and streptococcus faecium.
Summary of the invention
The deficiency and the defective that the present invention is directed to PFJ ensiling in the background technology make improvements, a kind of composite bacterial additive of alfalfa ensilage is provided, makes it in fermented green juice, add this streptococcus acidi lactici and streptococcus faecium and strengthened fermented green juice and promote ensiling clover fermentation, reduce the effect of pH value fast.
The present invention is achieved by the following technical solutions, the composite bacterial additive of alfalfa ensilage of the present invention is made up of streptococcus acidi lactici (Streptococcus lactis), streptococcus faecium (Streptococcus faecalis) and fermented green juice (PFJ), specifically be to mix with the weight percent of the MRS liquid nutrient medium of preserving streptococcus acidi lactici and streptococcus faecium by 50-60%: 20-25%: 20-25% by fermented green juice, form microoganism additives for ensiling lucerne, contain streptococcus acidi lactici or streptococcus faecium 5 * 10 at least in wherein every ml MRS liquid nutrient medium
6Cfu, this additive of every 10ml contains PFJ 5-6ml, streptococcus acidi lactici and streptococcus faecium each 10
6～10
7Cuf, the per kilogram clover adds the 10-15ml additive during ensiling.Fermented green juice and streptococcus acidi lactici are being raw material by the clover of ensiling, and through making and separating, streptococcus faecium separates from corn.Can adapt to the environment that alfalfa ensilage ferments by isolating streptococcus acidi lactici in the clover better than isolating streptococcus acidi lactici from other plant, produce more lactic acid.
Described MRS liquid nutrient medium (%) prescription is: peptone 1g, extractum carnis 1g, yeast extract paste 0.5g, K
2HPO
40.2g, dibasic ammonium citrate 0.2g, sodium acetate 0.5g, glucose 2g, soil temperature 800.1%, MgSO
4.7H
2O 0.058g, MnSO
4.4H
2O 0.025g, pH=6.2,121 ℃ of sterilization 15min.
Additive of the present invention makes by following steps: (1) makes fermented green juice; (2) separating lactic acid suis from fermented green juice; (3) streptococcus acidi lactici and streptococcus faecium activation; (4) fermented green juice and activatory streptococcus acidi lactici and streptococcus faecium are mixed in proportion, form the composite bacterial additive of alfalfa ensilage.
Characteristics of the present invention are: (1) streptococcus acidi lactici is by separating in the clover; (2) this streptococcus acidi lactici, streptococcus faecium and three kinds of additives that have complementary functions of fermented green juice are mixed in proportion, form this additive, it is different from external streptococcus acidi lactici or the PFJ method as addictive for alfalfa silage of using separately.Fermented green juice is the lactic acid fermented startup factor; Streptococcus acidi lactici is the lactic acid bacteria culturers that the preparation yeast phase plays a major role, and is the fastest lactic acid bacteria culturers of preparation yeast phase breeding; Streptococcus faecium is the lactic acid bacteria culturers that plays a major role the ensiling acidifying phase, it is the fastest lactic acid bacteria culturers of acidifying phase breeding, three 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 two kinds of additives of streptococcus acidi lactici.With the clover of this additive treating, through ensiling in 30 days, ensilage pH value reduced by 13.3～13.8% than handling with fermented green juice.
Embodiment
Provide following examples in conjunction with the inventive method step:
Embodiment one
1, fermented green juice is made: (1) gets 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, be chopped into 3-5cm, (2) add distilled water in clover (g) and 1: 2.5 ratio of water (ml), thorough mixing, 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 ℃ environment to ferment 48 hours, make fermented green juice;
2, streptococcus acidi lactici separation, purifying: with 100 times of fermented green juice dilutions, place the separate solid substratum, medium pH=5.8, cultivate 48h down for 35 ℃, select that molten calcium circle is big, the bacterium colony of substratum flavescence, be placed on MRS solid medium purifying, obtain streptococcus acidi lactici (ST), the streptococcus acidi lactici behind the purifying is preserved with the test tube that the MRS solid medium is housed.
Described separate solid substratum (%) prescription is: extractum carnis 1g, peptone 1g, yeast extract paste 1g, glucose 1g, tomato juice 20%, soil temperature 800.05%, lime carbonate 2g, tetrabromo-mcresolsulfonphthalein 0.01%, agar 1.5g, pH=5.8.
Described MRS solid medium (%) prescription is: peptone 1g, extractum carnis 1g, yeast extract paste 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.
3, streptococcus acidi lactici and streptococcus faecium activation: streptococcus acidi lactici and streptococcus faecium are placed in the MRS liquid nutrient medium activate respectively, 35 ℃ of activation temperatures, soak time 36 hours when activation is finished, contains streptococcus acidi lactici or streptococcus faecium 5 * 10 at least in every ml liquid nutrient medium
6Cfu.
Described MRS liquid nutrient medium (%) prescription is: peptone 1g, extractum carnis 1g, yeast extract paste 0.5g, K
2HPO
40.2g, dibasic ammonium citrate 0.2g, sodium acetate 0.5g, glucose 2g, soil temperature 800.1%, MgSO
4.7H
2O 0.058g, MnSO
4.4H
2O 0.025g, pH=6.2,121 ℃ of sterilization 15min.
4, the making of the composite bacterial additive of alfalfa ensilage: fermented green juice and the mixed of the MRS liquid nutrient medium of preserving streptococcus acidi lactici and streptococcus faecium by 50%: 25%: 25%, form microoganism additives for ensiling lucerne, wherein every this additive of 10ml contains PFJ 5ml, streptococcus acidi lactici and streptococcus faecium each 1.25 * 10
7Cuf, the per kilogram clover adds the 10ml additive during ensiling.
The alfalfa ensilage effect of table 2 embodiment one
| Handle | Sensible quality | pH | α＝0.05 | Protein | α＝0.05 |
| Additive-free this fermented liquid of PFJ | Inferior medium good | 5.10±0.12 4.87±0.16 4.30±0.21 | a b c | 17.25±0.51％ 20.31±0.12％ 23.26±0.32％ | c b a |
As seen from Table 2, with the clover of this fermentation liquor treatment, through ensiling in 30 days, the ensilage sensory evaluation reached good, and the sensory evaluation of PFJ is medium; Than PFJ processing low 13.3% and raising 14.5%, difference all reaches conspicuous level respectively for pH value and protein content.
Embodiment two
1, fermented green juice is made: (1) gets 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, be chopped into 3-5cm, (2) add distilled water in clover (g) and 1: 2.5 ratio of water (ml), thorough mixing, 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 ℃ environment to ferment 48 hours, make fermented green juice;
2, streptococcus acidi lactici separation, purifying: with 80 times of fermented green juice dilutions, place the separate solid substratum, medium pH=6.1, cultivate 48h down for 37 ℃, select that molten calcium circle is big, the bacterium colony of substratum flavescence, be placed on MRS solid medium purifying, obtain streptococcus acidi lactici (ST), the streptococcus acidi lactici behind the purifying is preserved with the test tube that the MRS solid medium is housed;
Described separate solid substratum (%) prescription is: extractum carnis 1g, peptone 1g, yeast extract paste 1g, glucose 1g, tomato juice 20%, soil temperature 800.05%, lime carbonate 2g, tetrabromo-mcresolsulfonphthalein 0.01%, agar 1.5g, pH=6.1.
Described MRS solid medium (%) prescription is identical with embodiment one.
3, streptococcus acidi lactici and streptococcus faecium activation: streptococcus acidi lactici and streptococcus faecium are placed in the MRS liquid nutrient medium activate respectively, 35 ℃ of activation temperatures, soak time 30 hours when activation is finished, contains streptococcus acidi lactici or streptococcus faecium 5 * 10 at least in every ml liquid nutrient medium
6Cfu.
Described MRS liquid nutrient medium (%) prescription is identical with embodiment one.
4, the making of the composite bacterial additive of alfalfa ensilage: fermented green juice and the mixed of the MRS liquid nutrient medium of preserving streptococcus acidi lactici and streptococcus faecium by 55%: 20%: 25%, form microoganism additives for ensiling lucerne, wherein every this additive of 10ml contains PFJ 5ml, streptococcus acidi lactici 10
7Cuf, streptococcus faecium 1.25 * 10
7Cuf, the per kilogram clover adds the 12ml additive during ensiling.
The alfalfa ensilage effect of table 3 embodiment one
| Handle | Sensible quality | pH | α＝0.05 | Protein | α＝0.05 |
| Additive-free this fermented liquid of PFJ | Inferior medium good | 5.10±0.12 4.87±0.16 4.28±0.13 | a b c | 17.25±0.51％ 20.31±0.12％ 23.49±0.71％ | c b a |
As seen from Table 3, with the clover of this fermentation liquor treatment, through ensiling in 30 days, the ensilage sensory evaluation reached good, and the sensory evaluation of PFJ is medium; Than PFJ processing low 13.8% and raising 15.7%, difference all reaches conspicuous level respectively for pH value and protein content.
Embodiment three
1, fermented green juice is made: (1) gets 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, be chopped into 3-5cm, (2) add distilled water in clover (g) and 1: 2.5 ratio of water (ml), thorough mixing, 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 ℃ environment to ferment 48 hours, make fermented green juice;
2, streptococcus acidi lactici 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 substratum flavescence, be placed on MRS solid medium purifying, obtain streptococcus acidi lactici (ST), the streptococcus acidi lactici behind the purifying is preserved with the test tube that the MRS solid medium is housed;
Described solid medium (%) prescription is: extractum carnis 1g, peptone 1g, yeast extract paste 1g, glucose 1g, tomato juice 20%, soil temperature 800.05%, lime carbonate 2g, tetrabromo-mcresolsulfonphthalein 0.01%, agar 1.5g, pH=6.4.
Described MRS solid medium (%) prescription is identical with embodiment one.
3, streptococcus acidi lactici and streptococcus faecium activation: streptococcus acidi lactici and streptococcus faecium are placed in the MRS liquid nutrient medium activate respectively, 37 ℃ of activation temperatures, soak time 24 hours when activation is finished, contains streptococcus acidi lactici or streptococcus faecium 5 * 10 at least in every m1 liquid nutrient medium
6Cfu.
Described MRS liquid nutrient medium (%) prescription is identical with embodiment one.
4, the making of the composite bacterial additive of alfalfa ensilage: fermented green juice and the mixed of the MRS liquid nutrient medium of preserving streptococcus acidi lactici and streptococcus faecium by 60%: 20%: 20%, form microoganism additives for ensiling lucerne, wherein every this additive of 10ml contains PFJ 6ml, streptococcus acidi lactici and streptococcus faecium each 10
7Cuf, the per kilogram clover adds the 15ml additive during ensiling.
The alfalfa ensilage effect of table 4 embodiment one
| Handle | Sensible quality | pH | α＝0.05 | Protein | α＝0.05 |
| Additive-free this fermented liquid of PFJ | Inferior medium good | 5.10±0.12 4.87±0.16 4.30±0.19 | a b c | 17.25±0.51％ 20.31±0.12％ 23.13±0.28％ | c b a |
As seen from Table 4, with the clover of this fermentation liquor treatment, through ensiling in 30 days, the ensilage sensory evaluation reached good, and the sensory evaluation of PFJ is medium; Than PFJ processing low 13.3% and raising 13.9%, difference all reaches conspicuous level respectively for pH value and protein content.
Claims (5)
1, a kind of composite bacterial additive of alfalfa ensilage is characterized in that, is mixed by the weight percent of 50-60%: 20-25%:20-25% by fermented green juice and the MRS liquid nutrient medium of preserving streptococcus acidi lactici and streptococcus faecium and forms.
2, the composite bacterial additive of alfalfa ensilage according to claim 1 is characterized in that, contains streptococcus acidi lactici or streptococcus faecium 5 * 10 in wherein every mlMRS liquid nutrient medium at least
6Cfu.
According to the composite bacterial additive of claim 1 or 2 described alfalfa ensilages, it is characterized in that 3, described MRS liquid culture based formulas is: peptone 1g, extractum carnis 1g, yeast extract paste 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.
4, the composite bacterial additive of alfalfa ensilage according to claim 1 is characterized in that, this additive of every 10ml contains fermented green juice 5-6ml, streptococcus acidi lactici and streptococcus faecium each 10
6～10
7Cuf.
According to the composite bacterial additive of claim 1 or 4 described alfalfa ensilages, it is characterized in that 5, the per kilogram clover adds the 10-15ml additive during ensiling.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100286955A CN1312274C (en) | 2005-08-11 | 2005-08-11 | Composite bacterial additive of alfalfa ensilage |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100286955A CN1312274C (en) | 2005-08-11 | 2005-08-11 | Composite bacterial additive of alfalfa ensilage |
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| Publication Number | Publication Date |
|---|---|
| CN1757714A true CN1757714A (en) | 2006-04-12 |
| CN1312274C CN1312274C (en) | 2007-04-25 |
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|---|---|---|---|
| CNB2005100286955A Expired - Fee Related CN1312274C (en) | 2005-08-11 | 2005-08-11 | Composite bacterial additive of alfalfa ensilage |
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Cited By (1)
| 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 |
-
2005
- 2005-08-11 CN CNB2005100286955A patent/CN1312274C/en not_active Expired - Fee Related
Cited By (2)
| 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 |
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| Publication number | Publication date |
|---|---|
| CN1312274C (en) | 2007-04-25 |
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