CN111685235B - Preparation method of lactobacillus plantarum fermented liquid complete feed for piglets - Google Patents
Preparation method of lactobacillus plantarum fermented liquid complete feed for piglets Download PDFInfo
- Publication number
- CN111685235B CN111685235B CN202010680995.6A CN202010680995A CN111685235B CN 111685235 B CN111685235 B CN 111685235B CN 202010680995 A CN202010680995 A CN 202010680995A CN 111685235 B CN111685235 B CN 111685235B
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- Prior art keywords
- lactobacillus plantarum
- feed
- liquid
- fermented liquid
- prepared
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- 239000007788 liquid Substances 0.000 title claims abstract description 69
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 57
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 56
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 56
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 239000000463 material Substances 0.000 claims abstract description 19
- 239000000843 powder Substances 0.000 claims description 31
- 238000000855 fermentation Methods 0.000 claims description 25
- 230000004151 fermentation Effects 0.000 claims description 25
- 239000001963 growth medium Substances 0.000 claims description 14
- 239000002994 raw material Substances 0.000 claims description 13
- 238000009630 liquid culture Methods 0.000 claims description 11
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 8
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- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims description 5
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- 244000068988 Glycine max Species 0.000 claims description 5
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- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims description 5
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- 239000001888 Peptone Substances 0.000 claims description 4
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- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 4
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- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
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- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 4
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- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims description 4
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- FUFJGUQYACFECW-UHFFFAOYSA-L calcium hydrogenphosphate Chemical compound [Ca+2].OP([O-])([O-])=O FUFJGUQYACFECW-UHFFFAOYSA-L 0.000 claims 1
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- 206010012735 Diarrhoea Diseases 0.000 abstract description 7
- 230000036541 health Effects 0.000 abstract description 5
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- 239000000126 substance Substances 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 description 27
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 22
- 239000004310 lactic acid Substances 0.000 description 11
- 235000014655 lactic acid Nutrition 0.000 description 11
- 241000894006 Bacteria Species 0.000 description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
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- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 4
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
- 239000010949 copper Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
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- 235000013305 food Nutrition 0.000 description 4
- 239000011572 manganese Substances 0.000 description 4
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
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- 238000004321 preservation Methods 0.000 description 3
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- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 description 2
- 240000000103 Potentilla erecta Species 0.000 description 2
- 235000016551 Potentilla erecta Nutrition 0.000 description 2
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 2
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- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 2
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- 229930003448 Vitamin K Natural products 0.000 description 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 2
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 2
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- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 2
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- 229910052742 iron Inorganic materials 0.000 description 2
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- 230000035764 nutrition Effects 0.000 description 2
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 2
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- 229920001817 Agar Polymers 0.000 description 1
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- 235000019750 Crude protein Nutrition 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- PNNCWTXUWKENPE-UHFFFAOYSA-N [N].NC(N)=O Chemical compound [N].NC(N)=O PNNCWTXUWKENPE-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
-
- 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
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/22—Animal feeding-stuffs from material of animal origin from fish
-
- 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
- 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
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- 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
- A23K20/142—Amino acids; Derivatives thereof
-
- 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
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
-
- 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
- A23K20/174—Vitamins
-
- 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
- A23K20/20—Inorganic substances, e.g. oligoelements
-
- 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
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/26—Compounds containing phosphorus
-
- 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
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/30—Oligoelements
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The invention relates to a preparation method of lactobacillus plantarum fermented liquid complete feed for piglets, which can effectively solve the problems of high feeding cost, poor gastrointestinal health, high diarrhea rate, poor palatability of traditional dry materials and low utilization rate of nutrient substances of the traditional weaned piglets, and the preparation method comprises the steps of inoculating lactobacillus plantarum into the prepared liquid complete feed, and fermenting at 160-200 rpm for 3-6 d at 26-33 ℃ to obtain the lactobacillus plantarum fermented liquid complete feed.
Description
Technical Field
The invention relates to the technical field of fermented feed, in particular to a preparation method of lactobacillus plantarum fermented liquid complete feed for piglets.
Background
In recent years, domestic food safety events frequently occur, in particular, drug residues and bacterial drug resistance are caused by abuse of antibiotics, food safety and human health are seriously threatened, and the safety of feeds is also receiving more and more attention from the nation and society. The microbial fermentation feed is taken as a novel green environment-friendly feed, and brings new research ideas for human beings. Therefore, under the new situation of advocating food safety and green cultivation, the development of fermented liquid feed is more required, and the safety of meat food is ensured from the source.
The fermented liquid feed (Fermented liquid Feed, FLF) is a feed raw material with good palatability and high nutrition level, which is produced by decomposing or converting anti-nutritional factors in the feed by beneficial microorganisms through metabolic activities under the artificial control of fermentation conditions. FLF has great advantages in application over traditional dry and liquid feeds: the FLF can greatly save feed cost, improve the digestion utilization rate of the feed, improve the growth performance of animals, generate a large amount of acidic substances in the liquid fermentation process, and strengthen the acidity of the feed, thereby inhibiting or killing salmonella and escherichia coli, finally promoting the health of animals, ensuring the health of human beings, and having good application prospect.
Disclosure of Invention
Aiming at the situation, the invention aims to solve the defects of the prior art, and the invention aims to provide a preparation method of lactobacillus plantarum fermented liquid complete feed for piglets, which can effectively solve the problems of high feeding cost, poor gastrointestinal health, high diarrhea rate, poor palatability, low nutrient utilization rate and the like of the traditional dry materials of the weaned piglets.
The invention solves the technical proposal that the lactobacillus plantarum UR-3 is classified and named as lactobacillus plantarum (Lactobacillus plantarum) which is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms for 12 months and 16 days in 2019, and has the preservation number of: CGMCC No.19131. Preservation address: no.1 and No. 3 of the north cinquefoil of the morning sun area of beijing city.
The method for fermenting liquid complete feed by using lactobacillus plantarum UR-3 is that lactobacillus plantarum is inoculated into the prepared liquid complete feed, the inoculum size is 4-6%, and the lactobacillus plantarum fermented liquid complete feed is obtained after fermentation for 3-6 d at 160-200 rpm of a shaking table at 26-33 ℃.
The complete feed is prepared from (by mass) corn 47-52%, soybean meal 16-24%, puffed soybean 16.5-20%, whey powder 1-4%, imported fish meal 1-5%, soybean oil 0.5-1.8%, lysine 0.1-0.14%, methionine 0.1-0.14%, salt 0.1-0.2%, stone powder 1.0-1.2%, calcium hydrophosphate 0.1-0.9% and premix 2.0%, wherein each kilogram of the premix is prepared from (copper) Cu 200mg, (iron) Fe 2300mg, (zinc) Zn2800 mg, (manganese) Mn 450mg, (vitamin A) VA 95000IU, (vitamin D) 3 )VD 3 13000IU, (vitamin E) VE 3000IU, (vitamin K) 3 )VK 3 90mg (vitamin B) 2 )VB 2 300mg and the balance of starch are uniformly mixed.
The lactobacillus plantarum is an excellent strain suitable for fermented liquid feed, can effectively inhibit the growth and propagation of mixed bacteria and harmful bacteria in feed raw materials, ensures good fermentation of the fermented liquid feed, has a simple process and easy operation, can be fermented in a rough pig farm environment, further enhances the intestinal health of piglets, accelerates the growth and weight gain of the piglets, maintains the normal physiological functions of organisms, improves the immune function of the piglets, has good effect, has obvious effect on the growth performance of the piglets, and is innovation in the fermented feed.
Detailed Description
The following describes the embodiments of the present invention in further detail with reference to examples.
Example 1
The invention relates to a method for preparing fermented liquid complete feed by using lactobacillus plantarum, which comprises the following specific steps:
1) Preparing seed liquid: inoculating Lactobacillus plantarum UR-3 into the prepared sterilized MRS liquid culture medium, fermenting at 160-180 rpm for 2-4 d at 26-33 ℃, then sucking 2-4% of strain, transferring into another bottle of newly prepared MRS liquid culture medium, fermenting at 160-180 rpm for 2-4 d at 26-33 ℃ to finally prepare Lactobacillus plantarum seed liquid with the viable count of the seed liquid being more than or equal to 1X 10 9 cfu/mL；
2) Pretreatment of raw materials: pulverizing the complete feed raw materials, fully and uniformly mixing, finely pulverizing the mixed raw materials to 60 meshes to prepare a fermented liquid feed dry powder material, and mixing the fermented liquid feed dry powder material according to a water-material ratio of 1: 2-3, and adding into a triangular flask;
3) Fermentation: inoculating the lactobacillus plantarum seed solution prepared in the step 1) into the triangular flask of the uniformly mixed and packaged dry powder material of the fermented liquid feed in the step 2), wherein the inoculum size is 4-6%, completely sealing, fermenting for 3-6 d at 160-200 rpm of a shaking table at 26-37 ℃, and ending the fermentation when the pH of the fermentation liquid is reduced to below 4.0, thus obtaining the lactobacillus plantarum fermented liquid complete feed.
Example 2
1) Preparing seed liquid: inoculating Lactobacillus plantarum UR-3 into the prepared sterilized MRS liquid culture medium, fermenting at 30deg.C with shaking table 170rpm for 3d, sucking 3% strain, transferring into another bottle of newly prepared MRS liquid culture medium, fermenting at 30deg.C with shaking table 170rpm for 3d, and making into Lactobacillus plantarum seed solution with viable count of not less than 1×10 9 cfu/mL；
2) Pretreatment of raw materials: 50% of corn, 20% of bean pulp, 19.01% of puffed soybean, 3% of whey powder, 3% of imported fish meal, 1% of soybean oil, 0.12% of lysine, 0.12% of methionine, 0.15% of salt, 1.1% of stone powder, 0.5% of calcium hydrophosphate and 2.0% of premix are crushed and fully and uniformly mixed, the mixed raw materials are finely crushed to 60 meshes, a fermented liquid feed dry powder material is prepared, and the fermented liquid feed dry powder material is prepared according to a water-material ratio of 1: 2-3, and adding into a triangular flask;
3) Fermentation: inoculating the lactobacillus plantarum seed solution prepared in the step 1) into the triangular flask of the uniformly mixed and packaged dry powder material of the fermented liquid feed in the step 2), wherein the inoculum size is 5%, completely sealing, fermenting for 4d at 180rpm of a shaking table at 30 ℃, and ending the fermentation when the pH of the fermentation liquid is reduced to below 4.0, thus obtaining the lactobacillus plantarum fermented liquid complete feed.
The MRS liquid culture medium is as follows: beef extract powder 5.0g, peptone 10.0g, yeast extract powder 4.0g, glucose 20.0g, triammonium citrate 2.0g, tween-80.0 ml, manganese sulfate 0.05g, magnesium sulfate 0.2g, sodium acetate 5.0g, dipotassium hydrogen phosphate 2.0g and distilled water 100ml, heating, boiling, dissolving and subpackaging, adjusting pH value to 6.2+/-0.2,121 ℃ and sterilizing for 15min for standby.
The premix is prepared from (copper) Cu 200mg, (iron) Fe 2300mg, (zinc) Zn2800 mg, (manganese) Mn 450mg, (vitamin A) VA 95000IU and (vitamin D) 3 )VD 3 13000IU, (vitamin E) VE 3000IU, (vitamin K) 3 )VK 3 90mg (vitamin B) 2 )VB 2 300mg and the balance of starch are uniformly mixed.
Detecting various indexes of lactobacillus plantarum fermented liquid complete feed, wherein the fermented feed product meets the technical standards of the product through index detection: the concentration of lactic acid is more than or equal to 150mmol/L, and the viable count of lactic acid bacteria is more than or equal to 1.0X10 9 cfu/mL, pH less than or equal to 4.5, acetic acid concentration less than or equal to 40mmol/L, acetic acid concentration less than or equal to 0.8mmol/L.
1. The screening method of the lactobacillus plantarum comprises the following steps:
1: uv mutagenesis of strains: preparing bacterial suspension from original bacterial strain (CGMCC No. 1.1856, lactobacillus plantarum Lactobacillus plantarum), diluting to different concentration gradients, coating on MRS solid culture medium after ultraviolet irradiation, wrapping with black cloth, culturing for 3-6 d at 26-33 ℃ in dark, taking out for counting, calculating the mortality, and finding out the optimal irradiation dose.
2: screening of mutagenized strains: (1) performing primary screening on a separation medium containing calcium carbonate through a calcium dissolving ring; (2) finally, a lactic acid high-yield strain UR-3 is obtained through re-screening, and the lactic acid yield is obviously improved.
The MRS solid culture medium is as follows: beef extract powder 5.0g, peptone 10.0g, yeast extract powder 4.0g, glucose 20.0g, triammonium citrate 2.0g, tween-80.0 ml, manganese sulfate 0.05g, magnesium sulfate 0.2g, sodium acetate 5.0g, agar powder 1.8g, dipotassium hydrogen phosphate 2.0g, calcium carbonate 2.0g and distilled water 100ml are heated, boiled, dissolved and packaged, and the pH value is adjusted to 6.2+/-0.2,121 ℃ for 15min for later use.
Compared with other strains of fermented liquid feed, the mutagenized strain UR-3 provided by the invention has good performances in the aspects of growth, lactic acid production and the like, and in the fermented liquid feed, the lactic acid production capacity of the UR-3 strain is improved by 44.53% compared with that of the original strain, the lactic acid concentration is more than or equal to 200mmol/L, the biomass is obviously improved, and the crude protein quantity is relatively improved.
3: identification of good strains:
picking the bacterial colony of the mutagenized excellent bacterial strain UR-3, streaking and inoculating the bacterial colony to an MRS culture medium plate for pure culture, culturing for 24-72 hours at 26-33 ℃, and carrying out microscopic examination to observe the color and morphological characteristics of the separated bacteria: forming white round raised colony with diameter of 1.0-3.0 mm, smooth and moist surface, regular edge, positive gram staining and no mobility. In combination with reference to the "Berger's bacteria identification handbook", wherein the results of the physiological and biochemical experiments of the strain are shown in Table 1, the results of the carbon source fermentation experiments are shown in Table 2, and the strain UR-3 is obtained by comparing the 16SrRNA sequencing data of the strain UR-3 according to the physiological and biochemical and carbon source fermentation experimental results, and the strain UR-3 is lactobacillus plantarum (Lactobacillus plantarum) which has been deposited in China general microbiological culture Collection center, with the accession number: CGMCC No.19131. Preservation address: no.1 and No. 3 of the north cinquefoil of the morning sun area of beijing city.
TABLE 1 physiological and biochemical experimental results of strain UR-3
Note that: "-" represents negative, "+" represents positive.
TABLE 2 fermentation test results of Strain UR-3 carbon source
Note that: "+" means strong fermentation, "-" means no fermentation, and "d" means slightly fermentation.
2. Feeding piglet experiments:
the lactobacillus plantarum fermented liquid pig complete feed provided by the invention is applied to feeding piglets, can obviously improve average daily gain and average daily feed intake of the piglets, reduce diarrhea rate and feed conversion ratio of the piglets, further improve blood biochemical indexes of the piglets, and is fully proved by test data, and related test data are as follows:
the lactobacillus plantarum fermented liquid pig complete feed provided by the invention is applied to feeding of piglets:
the 90 heads of the long and large ternary hybridization piglets of the age of 35 days are selected and randomly divided into 3 groups, and the test period is 21 days.
Test protocol
Group I: control group, feeding basic diet, the basic diet is: 50% of corn, 20% of soybean meal, 19.01% of puffed soybean, 3% of whey powder, 3% of imported fish meal, 1% of soybean oil, 0.12% of lysine, 0.12% of methionine, 0.15% of salt, 1.1% of stone powder, 0.5% of calcium hydrophosphate and 2.0% of premix.
Group II: feeding a basal diet containing an antimicrobial;
group III: feeding the lactobacillus plantarum fermented liquid complete feed obtained in the embodiment 2 of the invention.
The test group and the control group can eat and drink water freely, the test group and the control group weigh on an empty stomach at the beginning and the end of the test, the material consumption is counted, the diarrhea condition is observed, and the average daily gain, the food consumption and the diarrhea incidence rate are calculated.
The test results are shown in Table 1.
TABLE 1 Effect of lactic acid bacteria fermented liquid feeds on piglet production Performance
As can be seen from table 1: compared with the control group, the average daily gain of the test II group and the test III group is respectively improved by 17.68 percent (P < 0.05) and 26.88 percent (P < 0.05), the difference between the test II group and the test III group is not obvious (P > 0.05), the average daily feed intake of the test II group and the test III group is respectively improved by 5.34 percent (P < 0.05) and 10.68 percent (P < 0.05), and the feed conversion ratio of the test II group and the test III group is respectively reduced by 12.75 percent (P < 0.05) and 11.74 percent (P < 0.05). The lactobacillus plantarum fermented liquid feed can increase the daily feed intake of piglets and reduce diarrhea rate.
TABLE 2 influence of Lactobacillus plantarum fermented liquid pig complete feed on blood Biochemical index of piglets
As can be seen from Table 2, the total protein, immunoglobulin A and immunoglobulin G indices in the blood samples of test II and test III are significantly improved, the difference is significant (P < 0.05), the difference between test II and test III is significant (P < 0.05), the urea nitrogen content in the blood samples of test II and test III is significantly reduced, the difference is significant (P < 0.05), and the difference between test II and test III is significant (P < 0.05) compared with the control group.
Through repeated experiments in multiple batches, the products prepared by other embodiments of the invention can obtain the beneficial effects which are the same as or similar to the experimental results.
The lactobacillus plantarum fermented liquid complete feed provided by the invention is mainly applied to feeding piglets, the most difficult and complex link in the whole pig raising process is a piglet growth stage, and for piglets, the lactobacillus plantarum fermented liquid pig complete feed is relatively close to breast milk in form, can provide sufficient nutrition and water, and can be seen from feeding tests that the lactobacillus plantarum fermented liquid pig complete feed provided by the invention has good feeding effect, can improve the feed intake of piglets and reduce diarrhea rate; lactic acid generated in the fermentation process can inhibit the growth of intestinal pathogens, can obviously improve the immunity of piglets, effectively promote the healthy growth of the piglets, and is an ideal diet for the piglets; in addition, the related research on the influence of the lactobacillus plantarum fermented liquid pig complete feed on the piglet blood biochemical index is less, so that the research on the lactobacillus plantarum fermented liquid pig complete feed is significant to the development of the livestock industry in China, the research on the technology of the lactobacillus plantarum fermented liquid pig complete feed and the application of the lactobacillus plantarum fermented liquid pig complete feed on the piglet is a great innovation on the fermented feed, provides a test theoretical basis for replacing antibiotics in the antibiotic-free breeding application production of the lactobacillus plantarum fermented liquid pig complete feed, lays a theoretical foundation for further researching and developing safe, efficient and environment-friendly fermented feed, and has the potential of popularization and application in various pig farms as an efficient, green, ecological and safe feed.
The lactobacillus plantarum fermented liquid pig complete feed provided by the invention is used as a novel green environment-friendly feed, and has the advantages that: the feed is easy to produce and prepare, low in cost, safe, pollution-free, drug-free, digestible, small in damage to nutrient components of feed raw materials, good in application effect, completely meets the standard of fermented liquid feed products, contains rich lactic acid bacteria and metabolites thereof, lactic acid bacteria, small peptides, vitamins and the like, has sour and fragrant smell and good palatability, has a feeding promoting effect on animals, thereby increasing the feed intake of piglets, inhibiting the growth of intestinal harmful bacteria after being fed to livestock and poultry, maintaining the microecological balance of animal digestive tracts, enhancing the immunity of animal organisms, effectively preventing the occurrence of various diseases of the livestock and poultry, and is innovation in fermented feed, thereby having good popularization and application values.
Claims (3)
1. Lactobacillus plantarum UR-3, characterized in that it is designated lactobacillus plantarum (Lactobacillus plantarum) classified as deposited in the China Committee for culture Collection of microorganisms with deposit number: CGMCC No.19131.
2. Use of lactobacillus plantarum UR-3 as claimed in claim 1 for the preparation of liquid complete feed, in particularIs characterized in that lactobacillus plantarum is inoculated into prepared liquid complete feed, the inoculum size is 4 to 6 percent, and the lactobacillus plantarum fermented liquid complete feed is obtained after fermentation for 3 to 6 days at a temperature of 26 to 33 ℃ and a shaking table of 160 to 200 rpm; the complete feed is prepared from (by mass) corn 47-52%, soybean meal 16-24%, puffed soybean 16.5-20%, whey powder 1-4%, imported fish meal 1-5%, soybean oil 0.5-1.8%, lysine 0.1-0.14%, methionine 0.1-0.14%, salt 0.1-0.2%, stone powder 1.0-1.2%, calcium hydrogen phosphate 0.1-0.9% and premix 2.0%, wherein each kilogram of the premix is prepared from Cu 200 mg%, fe 2300mg, zn2800 mg, mn 450mg, VA 95000IU and VD 3 13000 IU、VE 3000 IU、VK 3 90 mg、VB 2 300 mixing mg and the balance of starch uniformly;
the method comprises the following steps:
1) Preparing seed liquid: inoculating Lactobacillus plantarum UR-3 into the prepared sterilized MRS liquid culture medium, fermenting at 160-180 rpm for 2-4 d at 26-33 ℃, then sucking 2-4% of strain, transferring into another bottle of newly prepared MRS liquid culture medium, fermenting at 160-180 rpm for 2-4 d at 26-33 ℃ to finally prepare Lactobacillus plantarum seed liquid with the viable count of the seed liquid being more than or equal to 1X 10 9 cfu/mL; the MRS liquid culture medium is as follows: beef extract powder 5.0g, peptone 10.0g, yeast extract powder 4.0g, glucose 20.0g, triammonium citrate 2.0g, tween-80.0 ml, manganese sulfate 0.05g, magnesium sulfate 0.2g, sodium acetate 5.0g, dipotassium hydrogen phosphate 2.0g and distilled water 100ml, heating, boiling, dissolving and subpackaging, adjusting pH value to 6.2+/-0.2,121 ℃ and sterilizing for 15min for standby;
2) Pretreatment of raw materials: pulverizing the complete feed raw materials, fully and uniformly mixing, finely pulverizing the mixed raw materials to 60 meshes to prepare a fermented liquid feed dry powder material, and mixing the fermented liquid feed dry powder material according to a water-material ratio of 1: 2-3, and adding into a triangular flask;
3) Fermentation: inoculating the lactobacillus plantarum seed solution prepared in the step 1) into the triangular flask of the uniformly mixed and packaged dry powder material of the fermented liquid feed in the step 2), wherein the inoculum size is 4-6%, completely sealing, fermenting for 3-6 d at 160-200 rpm of a shaking table at 26-37 ℃, and ending the fermentation when the pH of the fermentation liquid is reduced to below 4.0, thus obtaining the lactobacillus plantarum fermented liquid complete feed.
3. Use of lactobacillus plantarum UR-3 according to claim 2 for the preparation of liquid complete feed, characterized in that it comprises the following steps:
1) Preparing seed liquid: inoculating Lactobacillus plantarum UR-3 into the prepared sterilized MRS liquid culture medium, fermenting at 30deg.C with shaking table 170rpm for 3d, sucking 3% strain, transferring into another bottle of newly prepared MRS liquid culture medium, fermenting at 30deg.C with shaking table 170rpm for 3d, and making into Lactobacillus plantarum seed solution with viable count of not less than 1×10 9 cfu/mL; the MRS liquid culture medium is as follows: beef extract powder 5.0g, peptone 10.0g, yeast extract powder 4.0g, glucose 20.0g, triammonium citrate 2.0g, tween-80.0 ml, manganese sulfate 0.05g, magnesium sulfate 0.2g, sodium acetate 5.0g, dipotassium hydrogen phosphate 2.0g and distilled water 100ml, heating, boiling, dissolving and subpackaging, adjusting pH value to 6.2+/-0.2,121 ℃ and sterilizing for 15min for standby;
2) Pretreatment of raw materials: 50% of corn, 20% of bean pulp, 19.01% of puffed soybean, 3% of whey powder, 3% of imported fish meal, 1% of soybean oil, 0.12% of lysine, 0.12% of methionine, 0.15% of salt, 1.1% of stone powder, 0.5% of calcium hydrophosphate and 2.0% of premix are crushed and fully and uniformly mixed, the mixed raw materials are finely crushed to 60 meshes, a fermented liquid feed dry powder material is prepared, and the fermented liquid feed dry powder material is prepared according to a water-material ratio of 1: 2-3, and adding into a triangular flask;
3) Fermentation: inoculating the lactobacillus plantarum seed solution prepared in the step 1) into the triangular flask of the uniformly mixed and packaged dry powder material of the fermented liquid feed in the step 2), wherein the inoculum size is 5%, completely sealing, fermenting for 4d at 180rpm of a shaking table at 30 ℃, and ending the fermentation when the pH of the fermentation liquid is reduced to below 4.0, thus obtaining the lactobacillus plantarum fermented liquid complete feed.
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