CN1320106C - Preparation method of composite bacterial additive of alfalfa ensilage - Google Patents
Preparation method of composite bacterial additive of alfalfa ensilage Download PDFInfo
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- CN1320106C CN1320106C CNB200510028696XA CN200510028696A CN1320106C CN 1320106 C CN1320106 C CN 1320106C CN B200510028696X A CNB200510028696X A CN B200510028696XA CN 200510028696 A CN200510028696 A CN 200510028696A CN 1320106 C CN1320106 C CN 1320106C
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Abstract
The present invention relates to a method for preparing a composite bacterial additive of alfalfa ensilage, which belongs to the biotechnology field. The composite bacterial additive is formed by mixing 50 to 60 wt% of previously fermented juice and 20 to 25 wt% of MRS liquid culture medium in which streptococcus acidi lactici and streptococcus faecalis are preserved. The present invention comprises the following preparation steps: (1) making previously fermented juice; (2) extracting streptococcus acidi lactici from the previously fermented juice; (3) activating the streptococcus acidi lactici and streptococcus faecalis; (4) mixing the previously fermented juice and the activated streptococcus acidi lactici and the activated streptococcus faecalis according to proportion, and forming the composite bacterial additive of alfalfa ensilage. Compared with the effect achieved by singly using the two additives, namely the previously fermented juice and the streptococcus acidi lactici, the additive of the present invention has better effect of reducing pH and improving ensilage quality. After alfalfa processed with the additive is ensiled for 30 days, the pH value of the ensilage is 13.3 to 13.8% lower than processed with the previously fermented juice.
Description
Technical field
What the present invention relates to is a kind of method of biological technical field, specifically is a kind of preparation method 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 (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 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 and preparation method thereof 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 preparation method 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 described alfalfa ensilage 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 the MRS liquid nutrient medium of preserving streptococcus faecium and forms, and its preparation process is as follows:
(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.
Described step (1) is specially:
(a) 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, be chopped into 3-5cm;
(b) add distilled water in clover (g) and 1: 2.5 ratio of water (ml), thorough mixing with organizing pulverizer that mixed clover is smashed to pieces at a high speed, left standstill 1 hour, and double gauze filters;
(c) in filtrate, add the glucose or the suitable syrup of glucose amount in the ratio of clover raw material: glucose=10g: 1g, fully stir, place 30 ℃ environment fermentation 48 hours, make fermented green juice.
Described step (2) is specially: (a) with 60～100 times of fermented green juice dilutions, place the separate solid substratum, 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 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, and the product acidity of this streptococcus acidi lactici in 48 hours can see Table 1.
Described separate solid substratum (100ml) prescription is: extractum carnis 1g, peptone 1g, yeast extract paste 1g, glucose 1g, tomato juice 20%, soil temperature 80 0.05%, lime carbonate 2g, tetrabromo-mcresolsulfonphthalein 0.01%, agar 1.5g, pH5.8～6.4.
Described MRS solid medium (100ml) 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.
The pH value and the lactic acid content of table 1ST strain cultured solution change (mmol/L)
| 0h | 6h | 12h | 24h | 48h | |
| The pH lactic acid content | 6.09±0.02 0.50±0.03 | 4.87±0.04 3.30±0.03 | 4.34±0.02 17.74±0.43 | 3.99±0.01 27.91±1.30 | 4.02±0.01 36.35±1.29 |
Described step (3), be specially: streptococcus acidi lactici and streptococcus faecium are placed in the MRS liquid nutrient medium activate 35～37 ℃ of activation temperatures, soak time 24～36 hours respectively, when activation is finished, contain streptococcus acidi lactici or streptococcus faecium 5 * 10 in every ml liquid nutrient medium at least
6Cfu.
Described MRS liquid nutrient medium (100ml) 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, pH=6.2,121 ℃ of sterilization 15min.
Described step (4), be specially: fermented green juice with the MRS liquid nutrient medium of preserving streptococcus acidi lactici with preserve the mixed of the MRS liquid nutrient medium of streptococcus faecium by 50-60%: 20-25%: 20-25%, form microoganism additives for ensiling lucerne, wherein every this additive of 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.
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.At present, yet there are no fermented green juice is mixed report as addictive for alfalfa silage with streptococcus acidi lactici and streptococcus faecium.
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 (100ml) prescription is: extractum carnis 1g, peptone 1g, yeast extract paste 1g, glucose 1g, tomato juice 20%, soil temperature 80 0.05%, lime carbonate 2g, tetrabromo-mcresolsulfonphthalein 0.01%, agar 1.5g, pH=5.8.
Described MRS solid medium (100ml) 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 (100ml) 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, pH=6.2,121 ℃ of sterilization 15min.
4, the making of the composite bacterial additive of alfalfa ensilage: fermented green juice and the MRS liquid nutrient medium of preserving streptococcus acidi lactici and the mixed of the MRS liquid nutrient medium of preserving 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 (100ml) prescription is: extractum carnis 1g, peptone 1g, yeast extract paste 1g, glucose 1g, tomato juice 20%, soil temperature 80 0.05%, lime carbonate 2g, tetrabromo-mcresolsulfonphthalein 0.01%, agar 1.5g, pH=6.1.
Described MRS solid culture based formulas 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 MRS liquid nutrient medium of preserving streptococcus acidi lactici and the mixed of the MRS liquid nutrient medium of preserving 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 (100ml) 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 culture based formulas 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 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 MRS liquid nutrient medium of preserving streptococcus acidi lactici and the mixed of the MRS liquid nutrient medium of preserving 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± 2％ 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.
The making method flow process of above-mentioned three embodiment can be sketched and be:
Glucose → PFJ → the dilution of fermenting → make → streptococcus acidi lactici cultivation → purifying → acquisition streptococcus acidi lactici → choosing streptococcus faecium → activation → PFJ mixes with streptococcus acidi lactici and streptococcus faecium for fresh clover → selected → chopping → dipping → pulverizing (making beating) → leave standstill → filter → add → composite bacterial additive of alfalfa ensilage.
Claims (7)
1, a kind of preparation method of composite bacterial additive of alfalfa ensilage, it is characterized in that, 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 the MRS liquid nutrient medium of preserving streptococcus faecium and to form, preparation process is as follows:
(1) fermented green juice of making alfalfa;
(2) separating lactic acid suis from fermented green juice, wherein the isolation medium ph=5.8-6.4 of streptococcus acidi lactici;
(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.
2, the preparation method of the composite bacterial additive of alfalfa ensilage according to claim 1 is characterized in that, described step (1) is specially:
(a) 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, be chopped into 3-5cm;
(b) add distilled water, thorough mixing with organizing pulverizer that mixed clover is smashed to pieces at a high speed, left standstill 1 hour, and double gauze filters;
(c) in filtrate, add glucose, fully stir, place 30 ℃ environment to ferment 48 hours, make fermented green juice.
3, the preparation method of the composite bacterial additive of alfalfa ensilage according to claim 2 is characterized in that, in described (b), adds distilled water in clover g and 1: 2.5 ratio of water ml.
4, the preparation method of the composite bacterial additive of alfalfa ensilage according to claim 2 is characterized in that, in described (c), adds glucose in the ratio of clover raw material: glucose=10g: 1g in filtrate.
5, the preparation method of the composite bacterial additive of alfalfa ensilage according to claim 1 is characterized in that, described step (2) is specially:
(a) with 60～100 times of fermented green juice dilutions, place the separate solid substratum, 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 substratum flavescence, be placed on MRS solid medium purifying, obtain streptococcus acidi lactici.
6, the preparation method of the composite bacterial additive of alfalfa ensilage according to claim 1, it is characterized in that, described step (3), be specially: streptococcus acidi lactici and streptococcus faecium are placed in the MRS liquid nutrient medium activate respectively, 35～37 ℃ of activation temperatures, soak time 24～36 hours when activation is finished, contains streptococcus acidi lactici or streptococcus faecium 5 * 10 at least in every ml liquid nutrient medium
6Cfu.
7, the preparation method of the composite bacterial additive of alfalfa ensilage according to claim 1, it is characterized in that, described step (4), be specially: fermented green juice is mixed with the MRS liquid nutrient medium of preserving streptococcus acidi lactici and the weight percent of the MRS liquid nutrient medium of preserving streptococcus faecium by 50-60%: 20-25%: 20-25%, form microoganism additives for ensiling lucerne, wherein every this additive of 10ml contains fermented green juice 5-6ml, streptococcus acidi lactici and streptococcus faecium each 10
6～10
7Cuf, the per kilogram clover adds the 10-15ml additive during ensiling.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2159388A (en) * | 1984-06-01 | 1985-12-04 | Pan Britannica Ind Ltd | Preservation of silage |
-
2005
- 2005-08-11 CN CNB200510028696XA patent/CN1320106C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2159388A (en) * | 1984-06-01 | 1985-12-04 | Pan Britannica Ind Ltd | Preservation of silage |
Non-Patent Citations (4)
| Title |
|---|
| 优质青贮饲料发酵的调控 李槿年，饲料研究 1997 * |
| 绿汁发酵液和乳酸菌剂MMD3在不同含水率苜蓿青贮中的添加试验 张涛等，中国农业大学学报，第9卷第5期 2004 * |
| 绿汁发酵液和乳酸菌剂MMD3在不同含水率苜蓿青贮中的添加试验 张涛等，中国农业大学学报，第9卷第5期 2004;青贮添加剂的研究和应用 纪亚君，四川畜牧兽医，第1期 1996;优质青贮饲料发酵的调控 李槿年，饲料研究 1997 * |
| 青贮添加剂的研究和应用 纪亚君，四川畜牧兽医，第1期 1996 * |
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