CN114532445A - Additive for pasture silage - Google Patents
Additive for pasture silage Download PDFInfo
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- CN114532445A CN114532445A CN202210364152.4A CN202210364152A CN114532445A CN 114532445 A CN114532445 A CN 114532445A CN 202210364152 A CN202210364152 A CN 202210364152A CN 114532445 A CN114532445 A CN 114532445A
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- additive
- glucose
- culture medium
- pasture
- streptococcus
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- 239000000654 additive Substances 0.000 title claims abstract description 25
- 230000000996 additive effect Effects 0.000 title claims abstract description 19
- 239000004460 silage Substances 0.000 title abstract description 13
- 244000057717 Streptococcus lactis Species 0.000 claims abstract description 34
- 235000014897 Streptococcus lactis Nutrition 0.000 claims abstract description 34
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 28
- 239000008103 glucose Substances 0.000 claims abstract description 28
- 239000000284 extract Substances 0.000 claims abstract description 22
- 238000000855 fermentation Methods 0.000 claims abstract description 20
- 230000004151 fermentation Effects 0.000 claims abstract description 20
- 241000194032 Enterococcus faecalis Species 0.000 claims abstract description 18
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 244000025254 Cannabis sativa Species 0.000 claims abstract description 13
- 239000002994 raw material Substances 0.000 claims abstract description 6
- 244000283763 Acetobacter aceti Species 0.000 claims abstract description 4
- 235000007847 Acetobacter aceti Nutrition 0.000 claims abstract description 4
- 241000045403 Astragalus propinquus Species 0.000 claims abstract description 4
- 240000001046 Lactobacillus acidophilus Species 0.000 claims abstract description 4
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 claims abstract description 4
- 240000001929 Lactobacillus brevis Species 0.000 claims abstract description 4
- 235000013957 Lactobacillus brevis Nutrition 0.000 claims abstract description 4
- 244000199885 Lactobacillus bulgaricus Species 0.000 claims abstract description 4
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 claims abstract description 4
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 4
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 4
- 241000191998 Pediococcus acidilactici Species 0.000 claims abstract description 4
- 235000006533 astragalus Nutrition 0.000 claims abstract description 4
- 229940039695 lactobacillus acidophilus Drugs 0.000 claims abstract description 4
- 229940004208 lactobacillus bulgaricus Drugs 0.000 claims abstract description 4
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 4
- 239000001963 growth medium Substances 0.000 claims description 33
- 239000007787 solid Substances 0.000 claims description 22
- 230000004913 activation Effects 0.000 claims description 18
- 239000001888 Peptone Substances 0.000 claims description 15
- 108010080698 Peptones Proteins 0.000 claims description 15
- 235000015278 beef Nutrition 0.000 claims description 15
- 229940041514 candida albicans extract Drugs 0.000 claims description 15
- 235000019319 peptone Nutrition 0.000 claims description 15
- 239000012138 yeast extract Substances 0.000 claims description 15
- 238000009630 liquid culture Methods 0.000 claims description 14
- 239000000706 filtrate Substances 0.000 claims description 12
- 229920001817 Agar Polymers 0.000 claims description 10
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 10
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 10
- 239000008272 agar Substances 0.000 claims description 10
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 claims description 10
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 10
- 235000017281 sodium acetate Nutrition 0.000 claims description 10
- 239000001632 sodium acetate Substances 0.000 claims description 10
- 230000001954 sterilising effect Effects 0.000 claims description 10
- 238000001914 filtration Methods 0.000 claims description 9
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 8
- 229920000053 polysorbate 80 Polymers 0.000 claims description 8
- 238000004659 sterilization and disinfection Methods 0.000 claims description 7
- 239000009636 Huang Qi Substances 0.000 claims description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 6
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 6
- 239000002609 medium Substances 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- FRPHFZCDPYBUAU-UHFFFAOYSA-N Bromocresolgreen Chemical compound CC1=C(Br)C(O)=C(Br)C=C1C1(C=2C(=C(Br)C(O)=C(Br)C=2)C)C2=CC=CC=C2S(=O)(=O)O1 FRPHFZCDPYBUAU-UHFFFAOYSA-N 0.000 claims description 5
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 5
- CDUFCUKTJFSWPL-UHFFFAOYSA-L manganese(II) sulfate tetrahydrate Chemical compound O.O.O.O.[Mn+2].[O-]S([O-])(=O)=O CDUFCUKTJFSWPL-UHFFFAOYSA-L 0.000 claims description 5
- 235000015193 tomato juice Nutrition 0.000 claims description 5
- 230000001580 bacterial effect Effects 0.000 claims description 4
- 239000002689 soil Substances 0.000 claims description 4
- 235000019206 astragalus extract Nutrition 0.000 claims description 3
- 238000009835 boiling Methods 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 238000007865 diluting Methods 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 239000004459 forage Substances 0.000 claims description 3
- 238000005360 mashing Methods 0.000 claims description 3
- 238000000746 purification Methods 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 238000002791 soaking Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000006188 syrup Substances 0.000 claims description 3
- 235000020357 syrup Nutrition 0.000 claims description 3
- 210000001519 tissue Anatomy 0.000 claims description 3
- 238000009472 formulation Methods 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 238000004383 yellowing Methods 0.000 claims description 2
- 239000004461 grass silage Substances 0.000 claims 6
- 241000209504 Poaceae Species 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- 238000000034 method Methods 0.000 claims 1
- 238000002360 preparation method Methods 0.000 abstract description 6
- 241000411851 herbal medicine Species 0.000 abstract description 4
- 229920000136 polysorbate Polymers 0.000 description 3
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 150000003868 ammonium compounds Chemical class 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 235000005985 organic acids Nutrition 0.000 description 2
- 235000019629 palatability Nutrition 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 239000010902 straw Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000186660 Lactobacillus Species 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/113—Acidophilus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/121—Brevis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/123—Bulgaricus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/157—Lactis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/225—Faecalis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/231—Lactis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/41—Pediococcus
- A23V2400/413—Acidilactici
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Physiology (AREA)
- Molecular Biology (AREA)
- Health & Medical Sciences (AREA)
- Mycology (AREA)
- Botany (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses an additive for pasture silage, which comprises the following components: lactobacillus plantarum, Lactobacillus bulgaricus, Lactobacillus acidophilus, Acetobacter aceti, Lactobacillus brevis, Pediococcus acidilactici, lactococcus lactis subsp lactis and Astragalus membranaceus extract, wherein the Streptococcus lactis and Streptococcus faecalis need to be activated, and green juice fermentation broth needs to be used in the additive, wherein the green juice fermentation broth comprises pasture grass, water and glucose. The invention adds the Chinese herbal medicine preparation into the silage raw materials for fermentation, improves the silage quality and provides an effective way for the popularization and application of the Chinese herbal medicine.
Description
Technical Field
The invention relates to the technical field of pasture silage production, in particular to an additive for pasture silage.
Background
In order to expand the feed source and save the grain feed, nearly 6 hundred million tons of crop straws are produced every year in China, which is equivalent to dozens of times of wild hay produced by natural grasslands. But the straw feed has the disadvantages of coarse and hard texture, high lignified coarse cellulose content, poor palatability, low nutritional value and poor utilization effect. The silage is succulent feed prepared and stored by properly processing fresh green feed and fermenting by microorganisms such as lactobacillus under anaerobic condition, and not only can well keep the characteristics of green and succulent raw materials, but also has special sour aroma, soft texture, rich nutrition and good palatability.
However, the types of domestic and foreign silage additives mainly include ammonium compounds, organic acids, urea, biological preservatives and the like, while inorganic chemical additives have certain corrosivity and have potential safety hazards to people and livestock, while biological additives are imported from foreign countries and have high cost.
Disclosure of Invention
The invention aims to solve the following defects in the prior art, the types of silage additives at home and abroad mainly comprise ammonium compounds, organic acids, urea, biological preservatives and the like, but inorganic chemical additives have certain corrosivity and have potential safety hazards to people and livestock, while biological additives are imported from foreign countries and have higher cost.
In order to achieve the purpose, the invention adopts the following technical scheme:
an additive for the ensiling of pasture comprising:
lactobacillus plantarum, Lactobacillus bulgaricus, Lactobacillus acidophilus, Acetobacter aceti, Lactobacillus brevis, Pediococcus acidilactici, lactococcus lactis subsp lactis and Astragalus membranaceus extract, wherein the Streptococcus lactis and Streptococcus faecalis need to be activated.
Preferably, the additive requires the use of a green juice broth comprising grass, water and glucose.
Preferably, the preparation method of the fermentation liquor comprises the following steps:
(a) taking fresh pasture grass plants, removing the base and the top by about 10cm respectively, simultaneously removing main branches and thicker branches, reserving the remaining young branches and leaves, and chopping into 4 cm;
(b) pasture grass (g) and water (ml) 1: 2.5, adding distilled water, fully mixing, mashing the mixed pasture by using a high-speed tissue grinder, standing for 1 hour, and filtering by using double-layer gauze;
(c) according to the forage grass raw materials: glucose 10 g: adding glucose or syrup with the amount of glucose into the filtrate at a ratio of 1g, stirring, and fermenting at 30 deg.C for 48 hr to obtain green juice fermentation broth.
Preferably, the preparation method of the astragalus extract comprises the following steps:
(a) soaking radix astragali in water for 30min, boiling, decocting with slow fire, and filtering to obtain first filtrate and residue;
(b) adding 150ml of water into the obtained filter residue, decocting with slow fire, filtering for the second time, mixing the obtained second filtrate and the first filtrate, decocting with slow fire, and concentrating until the solid content is 0.96-1.04g/ml to obtain the radix astragali extract.
Preferably, the isolation procedure of the streptococcus lactis is as follows:
(a) diluting the green juice fermentation liquor by 60-100 times, placing the diluted green juice fermentation liquor in a separation solid culture medium, culturing the green juice fermentation liquor for 48 hours at 35-40 ℃, wherein the pH value of the culture medium is 5.8-6.4;
(b) selecting bacterial colony with large calcium-dissolving ring and yellowing culture medium, placing in MRS solid culture medium for purification to obtain Streptococcus lactis (ST), and storing the purified Streptococcus lactis in test tube filled with MRS solid culture medium.
Preferably, the formula of the isolated solid medium is as follows: 1-4g of beef extract, 1-4g of peptone, 1-4g of yeast extract, 1-4g of glucose, 20% of tomato juice, 800.05% of soil temperature, 2g of calcium carbonate, 0.01% of bromocresol green and 1.5g of agar, wherein the pH value is 5.8-6.4.
Preferably, the formulation of the MRS solid medium (%) is as follows: 1-4g of peptone, 1-4g of beef extract, 0-4g of yeast extract, 40.2K2HPO40, 0.2g of diammonium citrate, 0.5g of sodium acetate, 2g of glucose, 0.1% of Tween 80, 0.058g of MgSO4.7H2O, 0.025g of MnSO4.4H2O, 1.5g of agar, 6.2 of pH and 15min of sterilization at 121 ℃.
Preferably, the activation steps of the streptococcus lactis and streptococcus faecalis are as follows:
(a) respectively activating streptococcus lactis and streptococcus faecalis in an MRS liquid culture medium at the temperature of 35-37 ℃ for 24-36 hours;
(b) when the activation is completed, each ml of liquid culture medium at least contains 5 x 106cfu of streptococcus lactis or streptococcus faecalis, and the formula of the MRS liquid culture medium (%) is as follows: 1-4g of peptone, 1-4g of beef extract, 1-4g of yeast extract, 0-1g of K2HPO4, 0-1g of diammonium citrate, 0-1g of sodium acetate, 1-2g of glucose, 0.1% of soil temperature 80, 0.058g of MgSO4.7H2O, 0.025g of MnSO4.4Hx2O, 6.2 of pH, and sterilizing at 121 ℃ for 15 min.
Compared with the prior art, the invention has the beneficial effects that:
the Chinese herbal medicine preparation is added into the silage raw materials for fermentation, so that the silage quality is improved, and an effective way is provided for popularization and application of the Chinese herbal medicine.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
The first embodiment is as follows: an additive for pasture silage, the additive comprising:
lactobacillus plantarum, Lactobacillus bulgaricus, Lactobacillus acidophilus, Acetobacter aceti, Lactobacillus brevis, Pediococcus acidilactici, lactococcus lactis subsp lactis and Astragalus membranaceus extract, wherein the Streptococcus lactis and Streptococcus faecalis need to be activated.
In the preferred technical scheme of the embodiment, green juice fermentation liquor is required to be used in the additive, and the green juice fermentation liquor comprises pasture grass, water and glucose;
the preparation method of the fermentation liquor comprises the following steps:
(a) taking fresh pasture grass plants, removing the base and the top by about 10cm respectively, simultaneously removing main branches and thicker branches, reserving the remaining young branches and leaves, and chopping into 4 cm;
(b) pasture grass (g) and water (ml) 1: 2.5, adding distilled water, fully mixing, mashing the mixed pasture by using a high-speed tissue grinder, standing for 1 hour, and filtering by using double-layer gauze;
(c) according to the forage grass raw materials: glucose 10 g: adding glucose or syrup with the amount equivalent to that of glucose into the filtrate according to the proportion of 1g, fully stirring, and fermenting for 48 hours at the temperature of 30 ℃ to prepare green juice fermentation liquor;
the preparation method of the astragalus extract comprises the following steps:
(a) soaking radix astragali in water for 30min, boiling, decocting with slow fire, and filtering to obtain first filtrate and residue;
(b) adding 150ml of water into the obtained filter residue, decocting with slow fire, filtering for the second time, mixing the obtained second filtrate and the first filtrate, decocting with slow fire, and concentrating until the solid content is 0.96-1.04g/ml to obtain the radix astragali extract;
the separation steps of the streptococcus lactis are as follows:
(a) diluting the green juice fermentation broth by 60 times, placing in a solid medium with pH of 5.8, and culturing at 35 deg.C for 48 hr;
(b) selecting a bacterial colony which has a large calcium-dissolving ring and turns yellow in a culture medium, placing the bacterial colony in an MRS solid culture medium for purification to obtain streptococcus lactis (ST), and storing the purified streptococcus lactis in a test tube filled with the MRS solid culture medium;
the formula of the isolated solid culture medium is as follows: 1g of beef extract, 1g of peptone, 1g of yeast extract, 1g of glucose, 20% of tomato juice, 800.05% of Tween, 2g of calcium carbonate, 0.01% of bromocresol green and 1.5g of agar, wherein the pH is 5.87;
the MRS solid culture medium (%) comprises the following components: peptone 1, beef extract 1g, yeast extract 0-4g, K2HPO40.2g, diammonium citrate 0.2g, sodium acetate 0.5g, glucose 2g, Tween 80 0.1%, MgSO4.7H2O 0.058g, MnSO4.4H2O 0.025g, agar 1.5g, pH 6.2, and sterilization at 121 ℃ for 15 min;
the activation steps of the streptococcus lactis and the streptococcus faecalis are as follows:
(a) respectively putting the streptococcus lactis and the streptococcus faecalis in an MRS liquid culture medium for activation, wherein the activation temperature is 36 ℃, and the activation time is 30 hours;
(b) when the activation is completed, each ml of liquid culture medium at least contains 5 x 106cfu of streptococcus lactis or streptococcus faecalis, and the formula of the MRS liquid culture medium (%) is as follows: 1g of peptone, 1g of beef extract, 1g of yeast extract, 0-1g of K2HPO4, 0.2g of diammonium citrate, 0.2g of sodium acetate, 1g of glucose, 0.1% of Tween 80, 0.058g of MgSO4.7H2O, 0.025g of MnSO4.4Hx2O, 6.2 of pH, and sterilizing at 121 ℃ for 15 min.
Example two:
the formula of the isolated solid culture medium is as follows: 2g of beef extract, 2g of peptone, 2g of yeast extract, 2g of glucose, 20% of tomato juice, 800.05% of Tween, 2g of calcium carbonate, 0.01% of bromocresol green and 1.5g of agar, wherein the pH value is 6;
the MRS solid culture medium (%) comprises the following components: 2g of peptone, 2g of beef extract, 0.5g of yeast extract, 0.2g of K2HPO4, 0.2g of diammonium citrate, 0.5g of sodium acetate, 2g of glucose, 0.1% of Tween 80, 0.058g of MgSO4.7H2O, 0.025g of MnSO4.4H2O, 1.5g of agar, 6.2 of pH and 15min of sterilization at 121 ℃;
the activation steps of the streptococcus lactis and the streptococcus faecalis are as follows:
(a) respectively putting the streptococcus lactis and the streptococcus faecalis in an MRS liquid culture medium for activation, wherein the activation temperature is 3 ℃, and the activation time is 36 hours;
(b) when the activation is completed, each ml of liquid culture medium at least contains 5 x 106cfu of streptococcus lactis or streptococcus faecalis, and the formula of the MRS liquid culture medium (%) is as follows: 2g of peptone, 2g of beef extract, 2g of yeast extract, 2g of K2HPO4, 2g of diammonium citrate, 0.5g of sodium acetate, 0.5g of glucose, 0.1% of Tween 80, 0.058g of MgSO4.7H2O, 0.025g of MnSO4.4Hx2O, 6.2 of pH and 15min of sterilization at 121 ℃;
example three:
the formula of the isolated solid culture medium is as follows: 3g of beef extract, 3g of peptone, 3g of yeast extract, 3g of glucose, 20% of tomato juice, 800.05% of Tween, 2g of calcium carbonate, 0.01% of bromocresol green and 1.5g of agar, wherein the pH value is 6.4;
the MRS solid culture medium (%) comprises the following components: 3g of peptone, 3g of beef extract, 3g of yeast extract, 3g of K2HPO40.2g of diammonium citrate, 0.5g of sodium acetate, 2g of glucose, 0.1% of Tween 80, 0.058g of MgSO4.7H2O, 0.025g of MnSO4.4H2O, 1.5g of agar, 6.2 of pH and 15min of sterilization at 121 ℃;
the activation steps of the streptococcus lactis and the streptococcus faecalis are as follows:
(a) respectively putting the streptococcus lactis and the streptococcus faecalis into an MRS liquid culture medium for activation, wherein the activation temperature is 35-37 ℃, and the activation time is 24-36 hours;
(b) when the activation is completed, each ml of liquid culture medium at least contains 5 x 106cfu of streptococcus lactis or streptococcus faecalis, and the formula of the MRS liquid culture medium (%) is as follows: 4g of peptone, 4g of beef extract, 4g of yeast extract, 0-1g of K2HPO4, 1g of diammonium citrate, 0.7g of sodium acetate, 2g of glucose, 0.1% of Tween 80, 0.058g of MgSO4.7H2O, 0.025g of MnSO4.4Hx2O, 6.2 of pH and 15min of sterilization at 121 ℃.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical scope of the present invention by equivalent replacement or change according to the technical solution and the inventive concept of the present invention within the technical scope of the present invention.
Claims (8)
1. An additive for the ensiling of pasture grasses, the additive comprising:
lactobacillus plantarum, Lactobacillus bulgaricus, Lactobacillus acidophilus, Acetobacter aceti, Lactobacillus brevis, Pediococcus acidilactici, lactococcus lactis subsp lactis and Astragalus membranaceus extract, wherein the Streptococcus lactis and Streptococcus faecalis need to be activated.
2. The additive for pasture grass silage as claimed in claim 1, wherein the additive requires the use of green juice fermentation broth comprising pasture grass, water and glucose.
3. The green juice fermentation broth of claim 2 wherein said broth is prepared by:
(a) taking fresh pasture grass plants, removing the base and the top by about 10cm respectively, simultaneously removing main branches and thicker branches, reserving the remaining young branches and leaves, and chopping into 4 cm;
(b) pasture grass (g) and water (ml) 1: 2.5, adding distilled water, fully mixing, mashing the mixed pasture by using a high-speed tissue grinder, standing for 1 hour, and filtering by using double-layer gauze;
(c) according to the forage grass raw materials: glucose 10 g: adding glucose or syrup with the amount of glucose into the filtrate at a ratio of 1g, stirring, and fermenting at 30 deg.C for 48 hr to obtain green juice fermentation broth.
4. The additive for pasture grass silage as claimed in claim 1, wherein the astragalus extract is prepared by the following method:
(a) soaking radix astragali in water for 30min, boiling, decocting with slow fire, and filtering to obtain first filtrate and residue;
(b) adding 150ml of water into the obtained filter residue, decocting with slow fire, filtering for the second time, mixing the obtained second filtrate and the first filtrate, decocting with slow fire, and concentrating until the solid content is 0.96-1.04g/ml to obtain the radix astragali extract.
5. The additive for pasture grass silage as claimed in claim 1, wherein the streptococcus lactis is isolated by the following steps:
(a) diluting the green juice fermentation liquor by 60-100 times, placing the diluted green juice fermentation liquor in a separation solid culture medium, culturing the green juice fermentation liquor for 48 hours at 35-40 ℃, wherein the pH value of the culture medium is 5.8-6.4;
(b) selecting bacterial colony with large calcium-dissolving ring and yellowing culture medium, placing in MRS solid culture medium for purification to obtain Streptococcus lactis (ST), and storing the purified Streptococcus lactis in test tube filled with MRS solid culture medium.
6. The additive for pasture grass silage as claimed in claim 5, wherein the isolated solid medium formulation is: 1-4g of beef extract, 1-4g of peptone, 1-4g of yeast extract, 1-4g of glucose, 20% of tomato juice, 800.05% of soil temperature, 2g of calcium carbonate, 0.01% of bromocresol green and 1.5g of agar, wherein the pH value is 5.8-6.4.
7. The additive for pasture grass silage according to claim 5, wherein the MRS solid medium (%) formula is: 1-4g of peptone, 1-4g of beef extract, 0-4g of yeast extract, 40.2K2HPO40, 0.2g of diammonium citrate, 0.5g of sodium acetate, 2g of glucose, 0.1% of Tween 80, 0.058g of MgSO4.7H2O, 0.025g of MnSO4.4H2O, 1.5g of agar, 6.2 of pH and 15min of sterilization at 121 ℃.
8. The additive for pasture grass silage according to claim 1, wherein (a) streptococcus lactis and streptococcus faecalis are respectively activated in MRS liquid medium at 35-37 ℃ for 24-36 hours;
(b) when the activation is finished, each ml of liquid culture medium at least contains 5 x 106cfu of streptococcus lactis or streptococcus faecalis, and the formula of the MRS liquid culture medium (%) is as follows: 1-4g of peptone, 1-4g of beef extract, 1-4g of yeast extract, 0-1g of K2HPO4, 0-1g of diammonium citrate, 0-1g of sodium acetate, 1-2g of glucose, 0.1% of soil temperature 80, 0.058g of MgSO4.7H2O, 0.025g of MnSO4.4Hx2O, 6.2 of pH, and sterilizing at 121 ℃ for 15 min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210364152.4A CN114532445A (en) | 2022-04-08 | 2022-04-08 | Additive for pasture silage |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210364152.4A CN114532445A (en) | 2022-04-08 | 2022-04-08 | Additive for pasture silage |
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| CN202210364152.4A Pending CN114532445A (en) | 2022-04-08 | 2022-04-08 | Additive for pasture silage |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1757715A (en) * | 2005-08-11 | 2006-04-12 | 上海交通大学 | Preparation method of composite bacterial additive of alfalfa ensilage |
| CN103053808A (en) * | 2013-01-25 | 2013-04-24 | 中国农业大学 | Silage additive and preparation method and application thereof |
| CN105524867A (en) * | 2016-01-08 | 2016-04-27 | 中国农业大学 | Lactobacillus plantarum and application thereof in native grass silage |
| CN112515045A (en) * | 2020-11-30 | 2021-03-19 | 成都理工大学 | Silage additive for pasture silage |
-
2022
- 2022-04-08 CN CN202210364152.4A patent/CN114532445A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1757715A (en) * | 2005-08-11 | 2006-04-12 | 上海交通大学 | Preparation method of composite bacterial additive of alfalfa ensilage |
| CN103053808A (en) * | 2013-01-25 | 2013-04-24 | 中国农业大学 | Silage additive and preparation method and application thereof |
| CN105524867A (en) * | 2016-01-08 | 2016-04-27 | 中国农业大学 | Lactobacillus plantarum and application thereof in native grass silage |
| CN112515045A (en) * | 2020-11-30 | 2021-03-19 | 成都理工大学 | Silage additive for pasture silage |
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