CN101074426B - Method for producing bean-dregs feed containing conjugated linolic acid by plant lactobacillin fermentation - Google Patents
Method for producing bean-dregs feed containing conjugated linolic acid by plant lactobacillin fermentation Download PDFInfo
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- 230000004151 fermentation Effects 0.000 title claims abstract description 39
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- 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
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Abstract
A method for preparing a soybean meal forage rich in conjugated linoleic acid by fermenting vegetable lactobacillus belongs to the technical field of microorganism and animal nutrition and forage. The strain Lactobacillus plantarum ANCLA01 of the invention is preserved in the CGMCC with the preservation number of CGMCCNo.1906. The vegetable lactobacillus ANCLA01 is separated from corn silage, and is screened, purified and cultured with the name of Lactobacillus plantarum ANCLA01 applicable to soybean meal fermentation. The Lactobacillus plantarum ANCLA01 can improve the content of the conjugated linoleic acid in the ferment soybean meal forage and produce the functional ferment soybean meal forage rich in conjugated linoleic acid.
Description
Technical field
The invention belongs to microorganism and animal nutrition and feed technical field, be specifically related to the new bacterial strain of a kind of high yield conjugated linolic acid (CLA, Conjugated linoleic acid) and utilize its fermentative production to be rich in the functional bean-dregs feed of CLA.
Background technology
Utilize the domestic existing research of lactobacillus-fermented dregs of beans, but utilize the lactobacillus-fermented dregs of beans that produces CLA not report as yet.Anticancer, functions such as promotion is grown, inhibition accumulation of fat, enhancing body immunological competence that CLA has can improve pig, the fowl speed of growth and lean ratio, are the research focuses of subjects such as current medicine, protective foods and Animal nutrition.Milk and beef, mutton are human topmost CLA natural origins.People improve CLA content in milk from ruminants, the meat by two kinds of diet nutrient regulatory pathways: the one, and daily ration adds grease or changes daily ration adipose-derived, with the linoleic content of CLA metabolism substrate in the increase cud, and then improves CLA content in the livestock product; The 2nd, directly add CLA or ruminally-protected CLA (promptly crossing cud CLA).But because CLA costs an arm and a leg, it is too high to add cost in the feed, uses few in the production practice.Therefore, the animal nutrition and feed scholar is seeking other suitable, cost effective method that improves CLA content in the livestock product.
Summary of the invention
Technical problem to be solved by this invention is: the deficiency that is intended to overcome above-mentioned direct interpolation conjugated linolic acid and changes the daily ration lipid conformation, provide a kind of can the high yield conjugated linolic acid the new bacterial strain of plant lactobacillus and utilize its fermentative preparation to be rich in the method for conjugated linolic acid bean-dregs feed, to improve the content of conjugated linolic acid in the ruminant.
The new bacterial strain of plant lactobacillus of the present invention system is from separation from corn silage, screening, purifying and cultivate called after plant lactobacillus ANCLA01 (Lactobacillus plantarum ANCLA01).
The method that this plant lactobacillus fermentative preparation is rich in the conjugated linolic acid bean-dregs feed is: after earlier the strains A NCLA01 of this high yield conjugated linolic acid being spread cultivation in batches, after again the dregs of beans impurity elimination being pulverized, in bean cake powder, add wheat bran, carbohydrate, yeast, microcosmic salt, salt, magnesium salts, Vegetable oil lipoprotein and water, behind the mixing and stirring sterilization, plant lactobacillus ANCLA01 or plant lactobacillus ANCLA01 strain cultured solution are inoculated in the dregs of beans temperature controlled fermentation.
Description of drawings
Fig. 1 is the photo that plant lactobacillus ANCLAO1 of the present invention takes at microscopically;
Fig. 2 is a plant lactobacillus ANCLAO1 bacterial strain pcr amplification band photo of the present invention;
Fig. 3 is the ability of different strains bio-transformation conjugated linolic acid;
Fig. 4 is the utilization of plant lactobacillus ANCLAO1 to carbohydrate in the fermentation substrate.
Plant lactobacillus ANCLAO1 of the present invention (Lactobacillus plantarum ANCLAO1) has been carried out in accordance with regulations preservation: depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center abbreviates CGMCC as; Address: China. Beijing. Zhong Guan-cun, Institute of Microorganism, Academia Sinica; Preservation date: on January 8th, 2007; The numbering CGMCCNo.1906 that preservation is registered on the books.
Embodiment:
Embodiment: materials and methods
1.1 material: corn silage is collected in Hefei City white Supreme Being's dairy industry Development Co., Ltd; Substratum: pH6.5-6.8, the MRS broth culture, the PY substratum, MRS solid medium, MRS semisolid medium, MRS liquid nutrient medium and benefit are added 0.1g (100ml)
-1The MRS liquid nutrient medium of LA.Substratum is with the preceding 20min that all sterilizes under 121 ℃ of high pressure steam.
1.2 main agents: LA and CLA: purity 〉=99%, available from Sigma company; Bacterial genomes DNA extraction test kit, PCR test kit and glue reclaim test kit, available from precious biotechnology (Dalian) company limited; Other reagent is analytical pure.
1.3 bacterial screening:
1.3.1 primary dcreening operation: gather choose immediately after the ensiling Oranoleptic indicator preferably the ensiling branches and leaves immerse in 37 ℃ of distilled waters, remove branches and leaves after ten minutes, draw fresh even soak solution 10mL and be added to 500mL and be equipped with in the Erlenmeyer flask of 300mL MRS broth culture.37 ℃ static increase bacterium 96h after, dilution spread plate method is toppled over the MRS agar plate, cultivates 48h in 37 ℃ of anaerobism behind four rides, picking list bacterium colony percutaneous puncture-inoculation is cultivated rear defences for 37 ℃ and is preserved in refrigerator in the MRS semisolid medium.
1.3.2 multiple sieve: primary dcreening operation is preserved bacterial classification after 37 ℃ of activation, and the inoculum size by 5% is inoculated in the band nut test tube (d18mmx160mm) that 10mL MRS liquid nutrient medium is housed, and wherein linoleic acid content is 0.1g (100mL) in the MRS liquid nutrient medium
-1N
2Drive away air, measure CLA behind the anaerobically fermenting 24h, produce the CLA bacterium with screening.
1.4CLA measure: with the normal hexane is solvent, and the CLA standard specimen is made into the solution of different concns, is reference with the hexane, in 233nm place its absorption value of mensuration, is X-coordinate with CLA concentration (ug/mL), and light absorption value is an ordinate zou, the drawing standard curve.(1998) method operations such as Jiang are pressed in the extraction of CLA in the fermented liquid.Afterwards, institute is obtained fatty acid extract 5mL n-hexane dissolution, with the fermentation broth extract of not inoculating is reference, with ultraviolet spectrophotometer 00-350nm scope interscan again, read the absorption value at charateristic avsorption band 233nm place, calculate the content (Pariza etc., 1999) of CLA in the fermented liquid according to typical curve.Reference: substratum adds and the linolic acid of sample with amount, does not connect bacterial classification, the same sample of other step.
1.5 strain identification
1.5.1 bacterium colony and morphologic observation: after the superior strain activation that obtains, be inoculated on the MRS plate culture medium by four district's collimation methods, anaerobism is observed colony characteristics after cultivating 24h.Observe strain morphology feature, length range and width range etc. down in opticmicroscope behind the gramstaining.
1.5.2 Physiology and biochemistry is identified: with reference to " uncle's outstanding Bacteria Identification handbook the 9th edition (1994) is carried out experiments such as catalase reaction, gelatin liquification test, hydrogen sulfide reaction, indole reaction, temperature growth to the conjugated linolic acid superior strain that obtains and carried out preliminary evaluation.Produce evaluations of classifying of physiology such as acid, aerogenesis experiment and seminose, raffinose and biochemical aspect from glucose, experiment is with reference to " the 9th edition (1994) milk-acid bacteria classification evaluation of the outstanding Bacteria Identification handbook of uncle and experimental technique carry out concrete operations.
1.5.3 molecular biology identification: segmental recovery of purpose and purifying are all operated by the test kit description of step behind the pcr amplification of the preparation of template, bacterial strain 16SrDNA and the agarose gel electrophoresis.The purpose fragment that is recovered to is delivered to by precious biotechnology (Dalian) company limited then and checked order.With sequencing result use landed bacterium among BLAST software and the GenebanK the 16SrRNA sequence relatively, find and land of poor qualityly with the highest the landing the bacterial strain sequence number and access it of its similarity, carry out diversity ratio.
2. result
2.1 the screening of bacterial classification: when removing the cultivation of air conditions bottom fermentation, isolate 17 strains altogether and generate the stronger bacterial strain of CLA ability.By table 1 result as can be known, isolated bacterial strain CLA growing amount is between 4.316-33.442ug/mL (wet basis), and wherein No. 12 bacterial strain CLA output are up to 33.442ug/mL.This bacterial strain is named as ANCLA01.
2.2 strain identification:
2.2.1 the bacterium colony of bacterial classification and morphologic observation
Observe after the ANCLA01 bacterial strain MRS solid culture and find that bacterium colony projection, circle, smooth surface also are white in color; As shown in Figure 1, oily mirror is observed down and is found that the ANCLA01 bacterial strain is Gram-positive behind the gramstaining, short straight, the blunt circle in two ends of thalline, and no gemma, thalline is wide in 0.9～1.1um scope, and is long in 2～7um scope, becomes monomer, formation or short chain shape between thalline.Therefore, the ANCLA01 colonial morphology is with " the plant lactobacillus morphological specificity described in the outstanding Bacteria Identification handbook of uncle is more consistent.
2.2.2 Physiology and biochemistry check
Sugar fermentating test shows that ANCLA01 bacterial strain glucose fermentation result does not have gas and produces; Growth temperature detects finds that bacterial strain can be bred growth for 15 ℃, does not grow substantially for 45 ℃; Lactic acid generates and detects the test discovery, has lactic acid to generate in the fermented liquid.Take into account the above-mentioned bacterial strains morphological specificity, show this Pseudomonas homofermentation Type B lactobacillus.
2.2.2 molecular biology identification
The bacterial strain diversity ratio is found, 1482 base sequences identical (sequence has been uploaded GenBank, the number of landing EF185922) after 1482 base sequences that the 5th base of ANCLA01 bacterial strain 16S rRNA sequence is later and the 26th base of Lactobacillus plantarum strain L5.This result confirms that the ANCLA01 bacterial strain is strictly plant lactobacillus.
2.3 the different fermentations condition is to the influence of CLA output in the plant lactobacillus ANCLA01 fermented bean dregs
2.3.1 the wheat bran addition produces the influence of CLA to bean pulp fermentation
Because plant lactobacillus ANCLA01 is an amphimicrobe, add wheat bran and can increase the dregs of beans gap, help thalli growth.Press the 1t dregs of beans and add wheat bran 10,15,20,25,30 and the test of 35kg wheat bran, find behind the fermentation 60h: when the 1t dregs of beans added wheat bran 20kg, thalli growth was best, and CLA output is the highest, reaches 12.46mg/g (DW basis).
2.3.2 different carbon sources produce the influence of CLA to bean pulp fermentation
Though contain a small amount of sugar in the dregs of beans, far can not satisfy the microorganism growth needs.Based on dregs of beans, press the 1t dregs of beans and add starch 10,15,20,25,30 and 35kg starch test or 1t dregs of beans interpolation sucrose 10,15,20,25,30 and 35kg sucrose, find behind the fermentation 60h: when the 1t dregs of beans adds starch 25kg or sucrose 15kg, thalli growth is best, CLA output is the highest, reaches 14.53 mg/g (DW basis).
2.3.3. different phosphate sources is produced the influence of CLA to plant lactobacillus ANCLA01 fermented bean dregs
Phosphoric acid salt can cushion during the fermentation the medium pH value that the generation owing to lactic acid causes and descend, thereby helps the generation of CLA.Based on the bean pulp fermentation substratum, add at the 1t dregs of beans under the prerequisite of wheat bran 20kg, sucrose 10kg, yeast extract paste 15kg, NaCl1.5kg, add 15kg potassium hydrogen phosphate, potassium primary phosphate respectively, ammonium hydrogen phosphate, 6 kinds of microcosmic salt fermentations such as primary ammonium phosphate, Sodium phosphate dibasic, sodium hydrogen phosphate are found behind the fermentation 60h: add K
2HPO
4The time, thalli growth is best, and CLA output is the highest, reaches 16.21mg/g (DW basis).
2.3.4.K
2HPO
4Dosage is produced the influence of CLA to plant lactobacillus ANCLA01 fermented bean dregs
Based on the bean pulp fermentation substratum, add at the 1t dregs of beans under the prerequisite of wheat bran 20kg, sucrose 10kg, yeast extract paste 15kg, NaCl1.5kg, press the 1t dregs of beans and add starch 10,15,20,25,30 and 35kgK
2HPO
4, find behind the fermentation 60h: the 1t dregs of beans adds K
2HPO
4During 15kg, thalli growth is best, and CLA output is the highest, reaches 18.68mg/g (DW basis).
2.3.5. yeast extract paste is produced the influence of CLA to plant lactobacillus ANCLA01 fermented bean dregs
Lactobacillus plantarum is the class bacterial classification higher to nutritional requirement, and the complexity that nitrogenous source is utilized must influence elementary generation hilsa herring of this bacterium, thereby influences the output of CLA.CLA production needs somatomedins such as rare amino acid, and contains these somatomedins in the yeast extract paste, and therefore, an amount of yeast extract paste that adds helps thalli growth in substratum, helps the CLA ox and produces.Based on the bean pulp fermentation substratum, add wheat bran 20kg, sucrose 10kg, K at the 1t dregs of beans
2HPO
4Under the prerequisite of 15kg, NaCl1.5kg, press the 1t dregs of beans and add yeast extract paste 5,10,15,20,25,30kg, find behind the fermentation 60h: when the 1t dregs of beans added yeast extract paste 10kg, CLA output was 20.36mg/g (DW basis), strengthen the yeast extract paste addition, CLA output no longer obviously improves.
2.3.6. salt is produced the influence of CLA to plant lactobacillus ANCLA01 fermented bean dregs
Based on the bean pulp fermentation substratum, add wheat bran 20kg, sucrose 10kg, yeast extract paste 15kg, K at the 1t dregs of beans
2HPO
4Under the prerequisite of 15kg, press the 1t dregs of beans and add starch 0.5,1.0,1.5,2.0 and 2.5kg NaCl, find behind the fermentation 60h: when the 1t dregs of beans added NaCl1kg, thalli growth was best, and CLA output is the highest, reaches 16.13mg/g (DW basis).
2.3.7. magnesium salts is produced the influence of CLA to plant lactobacillus ANCLA01 fermented bean dregs
Based on the bean pulp fermentation substratum, add wheat bran 20kg, sucrose 10kg, yeast extract paste 15kg, K at the 1t dregs of beans
2HPO
4Under the prerequisite of 15kg, NaCl1.5kg, press interpolation 0.1,0.2,0.3,0.4 of 1t dregs of beans and 0.5kg MgSO
4, find behind the fermentation 60h: the 1t dregs of beans adds MgSO
4O.3kg the time, CLA reaches 22.41mg/g (DW basis).Strengthen MgSO
4Consumption, CLA output descends on the contrary.
2.3.8. fermentation time is produced the influence of CLA to plant lactobacillus ANCLA01 fermented bean dregs
Based on the bean pulp fermentation substratum, add wheat bran 20kg, sucrose 10kg, yeast extract paste 15kg, K at the 1t dregs of beans
2HPO
415kg, NaCl1.5kg, MgSO
40.3kg prerequisite under, fermentation time by 24,36,48,60,72,84,96h, found that: when fermenting after 48 hours, CLA no longer improves, and reaches 24.15mg/g (DW basis), shows that best fermentation time is 48h.
2.3.9. add Vegetable oil lipoprotein is produced CLA to plant lactobacillus ANCLA01 fermented bean dregs influence
Based on dregs of beans, add wheat bran 20kg, sucrose 10kg, yeast extract paste 15kg, K at the 1t dregs of beans
2HPO
415kg, NaCl1.5kg, MgSO
40.3kg prerequisite under, press the 1t dregs of beans and add sunflower seed oil 10,15,20,25,30,35kg, or the 1t dregs of beans adds rapeseed oil or peanut oil 10,15,20,25,30,35kg, find behind the fermentation 60h: the 1t dregs of beans adds sunflower seed oil 10kg, when rapeseed oil or peanut oil 30kg, CLA output is the highest, reaches 26.41mg/g (DW basis).
2.3.10. plant lactobacillus ANCLA01 inoculum size is produced the influence of CLA to plant lactobacillus ANCLA01 fermented bean dregs
Based on dregs of beans, add wheat bran 20kg, sucrose 10kg, yeast extract paste 15kg, K at the 1t dregs of beans
2HPO
415kg, NaCl1.5kg, MgSO
40.3kg, under the prerequisite of sunflower seed oil 10kg, plant lactobacillus ANCLA0l dosage of inoculation is by 6 * 10
7, 12 * 10
7, 18 * 10
7, 24 * 10
7, 30 * 10
7, 36 * 10
7Air-dry dregs of beans of CFU/g or plant lactobacillus ANCLA01 pure culture liquid addition are 150,300,450,600,750, the air-dry dregs of beans of 900g/t, plant lactobacillus ANCLA01 and pure culture liquid thereof with preceding must the dilution of sterilization warm water 10 times, be convenient to even inoculation.Fermentation 60h finds: 1t dregs of beans lactobacillus inoculum dosage is 12 * 10
7When air-dry dregs of beans of CFU/g or plant lactobacillus ANCLA0l pure culture liquid addition were the air-dry dregs of beans of 300g/t, CLA output was the highest, reaches 30.06mg/g (DW basis).
2.3.11. the fermentation condition combined effect is produced the influence of CLA to plant lactobacillus ANCLA01 fermented bean dregs
On the basis of above single factor experiment, pass through L
18(3
7) orthogonal optimization design investigates the entire effect of each factor to bean pulp fermentation.Found that the primary and secondary that influences each component of plant lactobacillus ANCLA01 generation CLA is followed successively by in proper order: starch or sucrose, K
2HPO
4, wheat bran, yeast extract paste, NaCl, MgSO
4, fermentation time, best of breed is: add wheat bran 20kg, starch 25kg or sucrose 15kg, yeast extract paste 10kg, microcosmic salt 15kg, salt 1kg, magnesium salts 0.3kg at the 1t dregs of beans, sunflower seed oil 10kg (or rapeseed oil, peanut oil 30kg), the lactobacillus inoculum amount is 12 * 10
7Air-dry dregs of beans of CFU/g or milk-acid bacteria pure culture liquid addition are the air-dry dregs of beans of 300g/t, and this moment, CLA output was up to 36.42mg/g (DW basis).
Utilize this plant lactobacillus fermentative preparation to be rich in the method for conjugated linolic acid bean-dregs feed:
1) plant lactobacillus ANCLA01 bacterial strain being carried out batch spreads cultivation: after will preserving twice of bacterial strain activation with above-mentioned MRS liquid nutrient medium, be inoculated in the 250mL triangular flask, and every bottled liquid measure 100mL, 37 ℃ of constant temperature are pressed 2x10 after leaving standstill and cultivating 48～72h again
7CFU/mL adds concentration and adds in the liquid seed fermentation jar, mixes back cultivation and fermentation 48h under 37 ℃ of conditions;
2) in bean-dregs feed, inoculate plant lactobacillus ANCLA01: after the dregs of beans impurity elimination is crushed to the 60-80 order, press 1t dregs of beans interpolation wheat bran 20kg, starch 25kg or sucrose 15kg, yeast extract paste 10kg, microcosmic salt 15kg, salt 1kg, magnesium salts 0.3kg, Vegetable oil lipoprotein (sunflower seed oil 10kg, rapeseed oil or peanut oil 30kg) 10-30kg, water 1t, the 20min that sterilizes under 115 ℃ of conditions after the mixing and stirring is when treating that temperature is reduced to 35-39 ℃, again by milk-acid bacteria dosage 12 * 10
7Air-dry dregs of beans of CFU/g or milk-acid bacteria pure culture liquid addition are the air-dry dregs of beans of 300g/t, (milk-acid bacteria and pure culture liquid thereof must dilute 10 times by sterilization warm water with preceding to spray inoculation plant lactobacillus ANCLA01 in dregs of beans, be convenient to even inoculation), anaerobically fermenting 48h under 35-39 ℃ of condition.Many batch fermentations are after survey CLA content all above more than the 30.52mg/g (DW basis).
Claims (5)
1. a plant lactobacillus fermentative preparation is rich in the method for conjugated linolic acid bean-dregs feed, it is characterized in that:
1) plant lactobacillus that is numbered CGMCCNo.1906 that preservation is registered on the books (Lactobacillus plantarum) ANCLA01 bacterial strain carries out batch and spreads cultivation: after will preserving bacterial strain and activate twice with the MRS liquid nutrient medium, be inoculated in the 250mL triangular flask, every bottled liquid measure 100mL, 37 ℃ of constant temperature are pressed 2x10 after leaving standstill and cultivating 48～72h again
7CFU/mL adds concentration and adds in the liquid seed fermentation jar, mixes back cultivation and fermentation 48h under 37 ℃ of conditions;
2) inoculation plant lactobacillus ANCLA01 in bean-dregs feed: back and wheat bran, sucrose or starch, yeast extract paste, microcosmic salt, salt, magnesium salts and Vegetable oil lipoprotein and water mixing and stirring are pulverized in the dregs of beans impurity elimination, after sterilizing, spray the above-mentioned also diluted plant lactobacillus ANCLA01 that spreads cultivation of inoculation, can enable behind the anaerobic condition bottom fermentation.
2. a kind of plant lactobacillus fermentative preparation according to claim 1 is rich in the method for conjugated linolic acid bean-dregs feed, it is characterized in that: described dregs of beans is 1: 0.02 with the mixed weight ratio of starch or sucrose, yeast extract paste, microcosmic salt, salt, magnesium salts and Vegetable oil lipoprotein and water: 0.015-0.025: 0.01: 0.015: 0.001: 0.0003: 0.01-0.03: 1.
3. a kind of plant lactobacillus fermentative preparation according to claim 1 is rich in the method for conjugated linolic acid bean-dregs feed, it is characterized in that: described Vegetable oil lipoprotein is sunflower seed oil or rapeseed oil or peanut oil.
4. a kind of plant lactobacillus fermentative preparation according to claim 1 is rich in the method for conjugated linolic acid bean-dregs feed, it is characterized in that: the dosage of described sprinkling inoculation plant lactobacillus ANCLA01 is by 12 * 10
7The air-dry dregs of beans of CFU/g.
5. a kind of plant lactobacillus fermentative preparation according to claim 4 is rich in the method for conjugated linolic acid bean-dregs feed, it is characterized in that: the described plant lactobacillus ANCLA01 that sprays inoculation in dregs of beans uses after diluting 10 times.
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| CN101020895B (en) * | 2007-01-24 | 2011-05-04 | 安徽农业大学 | Method for preparing silage rich in conjugated linoleic acid by lactobacillus plantarum fermentation |
| CN101525633B (en) * | 2009-04-13 | 2012-01-04 | 天津科技大学 | Method for preparing plant pathomycete zymotic fluid produced by lactobacillus |
| CN101632411B (en) * | 2009-08-20 | 2012-07-11 | 陈志敏 | Method for preparing high-quality rapeseed protein containing conjugated linoleic acid by multi-strain fermentation |
| CN102488085B (en) * | 2011-12-15 | 2013-12-11 | 安徽五粮泰生物工程股份有限公司 | Method of making fodder for young livestock |
| CN103205374B (en) * | 2013-03-15 | 2015-05-13 | 厦门和美科盛生物技术有限公司 | Lactic acid bacterium capable of reducing aflatoxin B1 in fermented feed and application of lactic acid bacterium |
| CN103478414B (en) * | 2013-09-11 | 2015-01-21 | 湖南大北农农业科技有限公司 | Feed rich in conjugated linoleic acid and preparation method of feed |
| CN103898028A (en) * | 2014-04-12 | 2014-07-02 | 东北农业大学 | High-density liquid-state cultivation method of feeding lactobacillus plantarum |
| CN106360077A (en) * | 2016-08-30 | 2017-02-01 | 惠州市埃德加科技有限公司 | Preparation method of nutrient foodstuff |
| CN107125453A (en) * | 2017-05-19 | 2017-09-05 | 魏聪 | A kind of preparation method for the fermented feed for raising snowflake beef cattle |
| CN114908017B (en) * | 2022-05-31 | 2023-03-14 | 四川农业大学 | Lactobacillus plantarum and application thereof in silage |
| CN115960767B (en) * | 2022-11-09 | 2024-04-26 | 厦门元之道生物科技有限公司 | Lactobacillus plantarum and application thereof |
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