CN105595012A - Method for preparing feed additive through compound microbial fermentation - Google Patents
Method for preparing feed additive through compound microbial fermentation Download PDFInfo
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- CN105595012A CN105595012A CN201610100497.3A CN201610100497A CN105595012A CN 105595012 A CN105595012 A CN 105595012A CN 201610100497 A CN201610100497 A CN 201610100497A CN 105595012 A CN105595012 A CN 105595012A
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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
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Abstract
The invention belongs to a method for preparing a feed additive through compound microbial fermentation. The method comprises the following steps that 1, water is added to a solid material according to the solid-liquid ratio of 2.5 kg: 1 L-5 kg: l L to obtain a solid-liquid mixture, and the solid-liquid mixture is sterilized and is cooled to reach 33-38 DEG C; 2, activated enterococcus faecalis is inoculated to a liquid enterococcus faecalis culture medium for cultivation; a bacillus subtilis seed solution is prepared: the activated bacillus subtilis is inoculated to a liquid bacillus subtilis culture medium for cultivation; a bacillus coagulans seed solution is prepared: activated bacillus coagulans is inoculated to a liquid bacillus coagulans culture medium for cultivation; 3, the bacillus subtilis seed solution, the bacillus subtilis seed solution and the bacillus coagulans seed solution prepared in the step 2 are mixed to obtain a hybrid seed solution, the hybrid seed solution is inoculated to the solid-liquid mixture obtained after sterilization in the step 1, and fermentation and drying are performed to obtain the feed additive.
Description
Technical field
The invention belongs to herb fermenting technical field of feed additive production, be specifically related to a kind of method that compound microorganism ferments is prepared feed addictive.
Background technology
Herb fermenting adopts microbial fermentation processes to produce Chinese medicine exactly. Chinese medicine preparation in traditional Chinese medicine is produced has just utilized natural microbial herb fermenting to improve herbal medicine efficacy, eliminate herbal toxic effect, to change herbal nature. Modern herb fermenting has adopted modern microbial fermentation technology, on purpose select microorganism and Chinese medicine collocation to carry out herb fermenting, its metabolism theory, effective component theory are different from traditional herb fermenting, and production controllability is good, and drug effect is higher, better, renewal.
Generally acknowledged Main Function mechanism has the following aspects at present: (1) drug effect is higher, and 100% absorbs: beneficial bacteria of intestinal tract fermented tcm, and the little molecularization of active ingredient, is easy to absorb; Medicinal fungi fermented tcm, active ingredient improves; Glucosides is transformed into aglycon by the beta-glucosidase of microorganism; The enzyme of microorganism makes the traditional Chinese medicine ingredients stripping in plant cell; (2) have no side effect, mouthfeel is better: the metabolite of sweat microorganism can divide toxolysin, can improve Chinese medicine mouthfeel, flavoring simultaneously; (3) drug effect is upgraded: the thalline of microorganism and metabolin, the microbial conversion of traditional Chinese medicine ingredients, the metabolic regulation effect of traditional Chinese medicine ingredients, interaction of microbe composition and traditional Chinese medicine ingredients etc., above all factors all can produce new effective component or modify new way is provided for active ingredient of Chinese herbs; (4) in Chinese medicine, also contain the nutritional labeling such as carbohydrate, protein, the digestive ferment that microbial metabolism produces can promote better digesting and assimilating of nutritional labeling; (5) drug effect improves, curative effect speed-raising. Compared with traditional Chinese medicine, after fermentation, the drug effect of Chinese medicine improves 4-28 doubly, and absorbs latter 30 minutes arrival blood.
Summary of the invention
The object of this invention is to provide a kind of compound microorganism ferments and prepare the method for feed addictive.
For achieving the above object, the technical solution used in the present invention is that a kind of compound microorganism ferments is prepared the method for feed addictive, comprise the following steps: 1. in solid material, add water by solid-to-liquid ratio 2.5kg ︰ 1L ~ 5kg ︰ 1L, sterilizing, is cooled to 33 ~ 38 DEG C, obtains solidliquid mixture; Wherein, described solid material is made up of the raw material of following portions by weight: Eucommia Leaf Powder 1-10 part, feed with paper-mulberry leaf powder 1-10 part, Astragalus Root P.E 1-5 part, wheat bran 1-3 part, calcium carbonate 0.05-0.3 part;
2. the preparation of enterococcus faecalis seed liquor: the enterococcus faecalis after activation is inoculated on enterococcus faecalis fluid nutrient medium, cultivates 24 hours for 37 ~ 39 DEG C; The preparation of bacillus subtilis seed liquor: the bacillus subtilis after activation is inoculated on bacillus subtilis fluid nutrient medium, cultivates 24 hours for 33 ~ 37 DEG C; The preparation of bacillus coagulans seed liquor: bacillus coagulans after activation is inoculated on bacillus coagulans fluid nutrient medium, cultivates 24 hours for 33 ~ 37 DEG C;
3. enterococcus faecalis seed liquor, bacillus subtilis seed liquor and bacillus coagulans seed liquor (1-10) by volume 2. step being prepared: (1-10): (1-10) mix, obtain seed mixture liquid, seed mixture liquid is inoculated in to the solidliquid mixture that 1. step obtains after sterilizing, temperature 35-40 DEG C of bottom fermentation 48-72 hour, be dried to water content < 6%, obtain feed addictive.
The formula of described enterococcus faecalis fluid nutrient medium is: peptone 30 ~ 50g/L, glucose 30 ~ 40g/L, salt 5 ~ 10g/L, hepar siccatum 5 ~ 7g/L, pH6.5; The formula of bacillus subtilis fluid nutrient medium is: yeast soaks powder 3-5g/L, glucose 10-20g/L, magnesium sulfate 3-5g/L, pH6.0; The formula of bacillus coagulans fluid nutrient medium is: yeast soaks powder 3-5g/L, tryptone 2-5g/L, glucose 10-20g/L, calcium chloride 3-5g/L, pH6.0.
Described step 1. in sterilizing be 121 DEG C of sterilizings 30 minutes in autoclave.
Described step 3. in the inoculation weight of seed mixture liquid be step 1. in solidliquid mixture weight 10 ~ 20%.
Described step 2. middle enterococcus faecalis activation is at 38 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula that enterococcus faecalis activates slant medium used is: peptone 30 ~ 50g/L, glucose 30 ~ 40g/L, salt 5 ~ 10g/L, hepar siccatum 5 ~ 7g/L, agar 15g/L, pH6.5; Bacillus subtilis activation is at 35 DEG C, to cultivate and activate for 24 hours on slant medium, the formula that bacillus subtilis activates slant medium used is: yeast soaks powder 3-5g/L, glucose 10-20g/L, magnesium sulfate 3-5g/L, agar 15-20g/L, pH6.0; Bacillus coagulans activation is at 35 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula of bacillus coagulans fluid nutrient medium is: yeast soaks powder 3-5g/L, tryptone 2-5g/L, glucose 10-20g/L, agar 15-20g/L, calcium chloride 3-5g/L, pH6.0.
Described Astragalus Root P.E can commercial existing Astragalus Root P.E; Preferably, the preparation method of described Astragalus Root P.E is as follows: in astragalus membranaceus powder, add water, the quality that adds of water is 8 times of astragalus membranaceus powder quality, floods 4 hours at 40-50 DEG C, and then refluxing extraction 2 hours, obtains filter residue and filtrate; In filter residue, add 6 times to the water of filter residue quality, refluxing extraction 2 hours, filter, then merge the filtrate obtaining for twice, being concentrated into density is 1.15-1.25g/ml, add ethanol to obtain mixture, in mixture, ethanol mass content is 75%, leaves standstill 8 hours, pumps supernatant, to precipitate and be dried, be yellow its extract.
In solid material, the contained glucide of the bark of eucommia and Astragalus Root P.E is as carbon source, protein in the bark of eucommia and paper mulberry and amino acid are as nitrogenous source, the vitamin that it is contained and carrotene are as growth factor, promote the growth of lactic acid bacteria (enterococcus faecalis, bacillus subtilis and bacillus coagulans), wheat bran can promote the output of Bacillus gemma.
The beneficial effect adopting in the present invention is: adopt flora combination in the present invention, its advantage of utilizing of novelty is abandoned its shortcoming. First, the metabolite peptide glycan of enterococcus faecalis has obvious curative effects to immunocompromised and RRTI etc., but enterococcus faecalis death rate after hydrochloric acid in gastric juice is higher, and bacillus coagulans stomach juice-resistant ability is strong, and has no drug resistance, good stability; Some lipase of bacillus subtilis energy metabolism, cellulase has good effect to the degree of depth degraded of solid material.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is further explained, but protection scope of the present invention is not limited in this. In following examples, enterococcus faecalis is purchased Bioteknologisk Institut in Beijing North Na Chuanlian, numbering LICC10319, and Latin formal name used at school:Enterococcusfaecalis; Bacillus subtilis is purchased in Chinese industrial microorganism fungus kind preservation administrative center, numbering CICC20445, and Latin formal name used at school:Bacillussubtilis; Bacillus coagulans is purchased in Qingdao DongHai Pharmacy Co., Ltd, and bacterial strain is bacillus coagulans TBC169, Latin formal name used at school:Bacilluscoagulans. Pork liver powder used is purchased from the biochemical Co., Ltd of Sanmenxia Gorge Bai Te.
Embodiment 1
Compound microorganism ferments is prepared a method for feed addictive, comprises the following steps: 1. in solid material, add water by solid-to-liquid ratio 4kg ︰ 1L, in autoclave, 121 DEG C of sterilizings 30 minutes, are cooled to 38 DEG C, obtain solidliquid mixture; Wherein, described solid material is made up of the raw material of following portions by weight: 5 parts of Eucommia Leaf Powders, 5 parts, feed with paper-mulberry leaf powder, 3 parts of Astragalus Root P.Es, 2 parts, wheat bran, 0.1 part, calcium carbonate;
2. the preparation of enterococcus faecalis seed liquor: the enterococcus faecalis after activation is inoculated on enterococcus faecalis fluid nutrient medium, cultivates 24 hours for 38 DEG C; The preparation of bacillus subtilis seed liquor: the bacillus subtilis after activation is inoculated on bacillus subtilis fluid nutrient medium, cultivates 24 hours for 35 DEG C; The preparation of bacillus coagulans seed liquor: bacillus coagulans after activation is inoculated on bacillus coagulans fluid nutrient medium, cultivates 24 hours for 35 DEG C;
3. enterococcus faecalis seed liquor, bacillus subtilis seed liquor and the bacillus coagulans seed liquor of 2. step being prepared by volume 5:5:4 mixed, obtain seed mixture liquid, by seed mixture liquid inoculation (the inoculation weight of seed mixture liquid be step 1. in solidliquid mixture weight 15%) solidliquid mixture that 1. obtains after sterilizing in step, 38 DEG C of bottom fermentations of temperature 48 hours, in drying box, at 43 DEG C, be dried to water content < 5%, obtain feed addictive.
The formula of described enterococcus faecalis fluid nutrient medium is: peptone 40g/L, glucose 35g/L, salt 8g/L, pork liver powder 6g/L, pH6.5; The formula of bacillus subtilis fluid nutrient medium is: yeast soaks powder 5g/L, glucose 15g/L, magnesium sulfate 5g/L, pH6.0; The formula of bacillus coagulans fluid nutrient medium is: yeast soaks powder 3g/L, tryptone 2g/L, glucose 15g/L, calcium chloride 5g/L, pH6.0.
Described step 2. middle enterococcus faecalis activation is at 38 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula that enterococcus faecalis activates slant medium used is: peptone 40g/L, glucose 35g/L, salt 8g/L, pork liver powder 6g/L, agar 15g/L, pH6.5; Bacillus subtilis activation is at 35 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula that bacillus subtilis activates slant medium used is: yeast soaks powder 5g/L, glucose 15g/L, magnesium sulfate 5g/L, agar 20g/L, pH6.0; ; Bacillus coagulans activation is at 35 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula of bacillus coagulans fluid nutrient medium is: yeast soaks powder 3g/L, tryptone 2g/L, glucose 15g/L, agar 18g/L, calcium chloride 5g/L, pH6.0.
The preparation method of described Astragalus Root P.E is as follows: in astragalus membranaceus powder, add water, the quality that adds of water is 8 times of astragalus membranaceus powder quality, floods 4 hours at 45 DEG C, and then refluxing extraction 2 hours, obtains filter residue and filtrate; In filter residue, add 6 times to the water of filter residue quality, refluxing extraction 2 hours, filter, then merge the filtrate obtaining for twice, being concentrated into density is 1.2g/ml, add ethanol to obtain mixture, in mixture, ethanol mass content is 75%, leaves standstill 8 hours, pumps supernatant, to precipitate and be dried, be yellow its extract.
Embodiment 2
Compound microorganism ferments is prepared a method for feed addictive, comprises the following steps: 1. in solid material, add water by solid-to-liquid ratio 2.5kg ︰ 1L, in autoclave, 121 DEG C of sterilizings 30 minutes, are cooled to 35 DEG C, obtain solidliquid mixture; Wherein, described solid material is made up of the raw material of following portions by weight: 1 part of Eucommia Leaf Powder, 10 parts, feed with paper-mulberry leaf powder, 1 part of Astragalus Root P.E, 3 parts, wheat bran, 0.3 part, calcium carbonate;
2. the preparation of enterococcus faecalis seed liquor: the enterococcus faecalis after activation is inoculated on enterococcus faecalis fluid nutrient medium, cultivates 24 hours for 37 DEG C; The preparation of bacillus subtilis seed liquor: the bacillus subtilis after activation is inoculated on bacillus subtilis fluid nutrient medium, cultivates 24 hours for 33 DEG C; The preparation of bacillus coagulans seed liquor: bacillus coagulans after activation is inoculated on bacillus coagulans fluid nutrient medium, cultivates 24 hours for 33 DEG C;
3. enterococcus faecalis seed liquor, bacillus subtilis seed liquor and the bacillus coagulans seed liquor of 2. step being prepared by volume 1:10:1 mixed, obtain seed mixture liquid, by seed mixture liquid inoculation (the inoculation weight of seed mixture liquid be step 1. in solidliquid mixture weight 20%) solidliquid mixture that 1. obtains after sterilizing in step, 35 DEG C of bottom fermentations of temperature 72 hours, in air dry oven, at 40 DEG C, be dried to water content < 5%, obtain feed addictive.
The formula of described enterococcus faecalis fluid nutrient medium is: peptone 30g/L, glucose 40g/L, salt 5g/L, pork liver powder 5g/L, pH6.5; The formula of bacillus subtilis fluid nutrient medium is: yeast soaks powder 3g/L, glucose 20g/L, magnesium sulfate 4g/L, pH6.0; The formula of bacillus coagulans fluid nutrient medium is: yeast soaks powder 5g/L, tryptone 3g/L, glucose 10g/L, calcium chloride 3g/L, pH6.0.
Described step 2. middle enterococcus faecalis activation is at 38 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula that enterococcus faecalis activates slant medium used is: peptone 30g/L, glucose 40g/L, salt 5g/L, pork liver powder 5g/L, agar 20g/L, pH6.5; Bacillus subtilis activation is at 35 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula that bacillus subtilis activates slant medium used is: yeast soaks powder 3g/L, glucose 20g/L, magnesium sulfate 4g/L, agar 15g/L, pH6.0; ; Bacillus coagulans activation is at 35 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula of bacillus coagulans fluid nutrient medium is: yeast soaks powder 5g/L, tryptone 3g/L, glucose 10g/L, agar 15g/L, calcium chloride 3g/L, pH6.0.
The preparation method of described Astragalus Root P.E is as follows: in astragalus membranaceus powder, add water, the quality that adds of water is 8 times of astragalus membranaceus powder quality, floods 4 hours at 40 DEG C, and then refluxing extraction 2 hours, obtains filter residue and filtrate; In filter residue, add 6 times to the water of filter residue quality, refluxing extraction 2 hours, filter, then merge the filtrate obtaining for twice, being concentrated into density is 1.15g/ml, add ethanol to obtain mixture, in mixture, ethanol mass content is 75%, leaves standstill 8 hours, pumps supernatant, to precipitate and be dried, be yellow its extract.
Embodiment 3
Compound microorganism ferments is prepared a method for feed addictive, comprises the following steps: 1. in solid material, add water by solid-to-liquid ratio 5kg ︰ 1L, in autoclave, 121 DEG C of sterilizings 30 minutes, are cooled to 33 DEG C, obtain solidliquid mixture; Wherein, described solid material is made up of the raw material of following portions by weight: 10 parts of Eucommia Leaf Powders, 1 part, feed with paper-mulberry leaf powder, 5 parts of Astragalus Root P.Es, 1 part, wheat bran, 0.05 part, calcium carbonate;
2. the preparation of enterococcus faecalis seed liquor: the enterococcus faecalis after activation is inoculated on enterococcus faecalis fluid nutrient medium, cultivates 24 hours for 39 DEG C; The preparation of bacillus subtilis seed liquor: the bacillus subtilis after activation is inoculated on bacillus subtilis fluid nutrient medium, cultivates 24 hours for 37 DEG C; The preparation of bacillus coagulans seed liquor: bacillus coagulans after activation is inoculated on bacillus coagulans fluid nutrient medium, cultivates 24 hours for 37 DEG C;
3. enterococcus faecalis seed liquor, bacillus subtilis seed liquor and the bacillus coagulans seed liquor of 2. step being prepared by volume 10:1:10 mixed, obtain seed mixture liquid, by seed mixture liquid inoculation (the inoculation weight of seed mixture liquid be step 1. in solidliquid mixture weight 10%) solidliquid mixture that 1. obtains after sterilizing in step, 40 DEG C of bottom fermentations of temperature 60 hours, on fluid bed, at 44 DEG C, be dried to water content 65%, obtain feed addictive.
The formula of described enterococcus faecalis fluid nutrient medium is: peptone 50g/L, glucose 30g/L, salt 10g/L, pork liver powder 7g/L, pH6.5; The formula of bacillus subtilis fluid nutrient medium is: yeast soaks powder 4g/L, glucose 10g/L, magnesium sulfate 3g/L, pH6.0; The formula of bacillus coagulans fluid nutrient medium is: yeast soaks powder 4g/L, tryptone 5g/L, glucose 20g/L, calcium chloride 4g/L, pH6.0.
Described step 2. middle enterococcus faecalis activation is at 38 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula that enterococcus faecalis activates slant medium used is: peptone 50g/L, glucose 30g/L, salt 10g/L, pork liver powder 7g/L, agar 18g/L, pH6.5; Bacillus subtilis activation is at 35 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula that bacillus subtilis activates slant medium used is: yeast soaks powder 4g/L, glucose 10g/L, magnesium sulfate 3g/L, agar 18g/L, pH6.0; Bacillus coagulans activation is at 35 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula of bacillus coagulans fluid nutrient medium is: yeast soaks powder 4g/L, tryptone 5g/L, glucose 20g/L, agar 20g/L, calcium chloride 4g/L, pH6.0.
The preparation method of described Astragalus Root P.E is as follows: in astragalus membranaceus powder, add water, the quality that adds of water is 8 times of astragalus membranaceus powder quality, floods 4 hours at 50 DEG C, and then refluxing extraction 2 hours, obtains filter residue and filtrate; In filter residue, add 6 times to the water of filter residue quality, refluxing extraction 2 hours, filter, then merge the filtrate obtaining for twice, being concentrated into density is 1.25g/ml, add ethanol to obtain mixture, in mixture, ethanol mass content is 75%, leaves standstill 8 hours, pumps supernatant, to precipitate and be dried, be yellow its extract.
Application test 1
On December 1st, 2013---on January 15th, 2014 is on pig farm, Sanmenxia Gorge Lingbao City, chooses totally 4 of the Landraces of first 30 days of the expected date of childbirth, is numbered 1,2,3, No. 4, is divided into two groups, and No. 1, No. 2 is test group, and No. 3, No. 4 is control group.
Test group: No. 1, No. 2 in-pigs first 30 days in the expected date of childbirth, 1% added embodiment 3 gained feed addictives in mixed fodder by mass percentage December 1, the sow of feeding, until stop using when farrowing.
Control group: No. 3, No. 4 in-pigs identical mixed fodder of yet as usual feeding from December 1, does not add any medicine and additive.
Two groups of tests, contrast all under certain environmental condition, are carried out feeding and management by a special poultry raiser, and the number of times of feeding, method, drinking-water require basically identical, require keeper to make a record. Test group and control group are all observed one and a half months, record litter size, incidence.
Test group is in 13 of No. 1 pig farrowing on January 2nd, 2014,12 of No. 2 pig farrowing, totally 25; Within 2014, in 9 of No. 3 pig farrowing on January 2, No. 2 pig farrows 8 control group, totally 17. In postpartum first quarter moon, the whole normal growths of test group piglet, 5 of control group morbidities.
Through test of many times, first 30 days of the expected date of childbirth was used feed addictive, can obviously improve farrowing rate 20-30%, and piglet all can healthy growth.
Application test 2
On June 1st, 2014---on September 30th, 2014 is in chicken farm, Sanmenxia Gorge Lingbao City, choose 1 the monthly age 400 of soil chickens, be divided at random two groups, each 200 of test group, control group.
Test group: June 1 was that 3% interpolation embodiment 1 gained feed addictive is fed in mixed fodder by mass percentage, until deliver for sale.
Control group: the identical mixed fodder of yet as usual feeding from June 1, does not add any medicine and additive.
Two groups of tests, contrast all under certain environmental condition, are carried out feeding and management by a special poultry raiser, and the number of times of feeding, method, drinking-water require basically identical, require keeper to make a record, and after delivering for sale, butcher and check chicken.
Test group is at duration of test, and the incidence of disease is obviously less than control group; In test group cock skin, hydroxyproline content reaches 26.27%, high control group 50.29%. While butchering, the toughness of chicken strengthens, the muscle fibre prosperity of thigh position, obviously chap, myocyte's gap turn narrow; In chicken, cholesterol and fat content all have decline in various degree.
Application test 3
On May 1st, 2015---on August 30th, 2015, in chicken farm, Sanmenxia Gorge Lingbao City, is chosen 400 of laying hens, is divided at random two groups, and test group, control group are respectively 200.
Test group: May 1,1% interpolation embodiment 2 gained feed addictives were fed in mixed fodder by mass percentage, until deliver for sale.
Control group: the identical mixed fodder of yet as usual feeding from May 1, does not add any medicine and additive.
Two groups of tests, contrast all under certain environmental condition, are carried out feeding and management by a special poultry raiser, and the number of times of feeding, method, drinking-water require basically identical, require keeper to make a record, and regularly add up laying rate, periodic inspection egg quality.
Test group is fed egg cholesterol after 30 days and is reduced by 6.05% ~ 10.45%, feeds and reduces by 6.92% ~ 14.29% after 60 days, feeds and reduces by 11.60% ~ 17.65% after 90 days, and chicken is fed egg inner cholesterol content after 4 months and reduces by 14.97% ~ 20.69%; Laying rate is slightly improved compared with control group, but increase rate is little; The difference of test group and single piece of weight of egg of control group is not obvious.
Claims (7)
1. compound microorganism ferments is prepared a method for feed addictive, it is characterized in that, comprises the following steps:
1. in solid material, add water by solid-to-liquid ratio 2.5kg ︰ 1L ~ 5kg ︰ 1L, sterilizing, is cooled to 33 ~ 38 DEG C, obtains solidliquid mixture; Wherein, described solid material is made up of the raw material of following portions by weight: Eucommia Leaf Powder 1-10 part, feed with paper-mulberry leaf powder 1-10 part, Astragalus Root P.E 1-5 part, wheat bran 1-3 part, calcium carbonate 0.05-0.3 part;
2. the preparation of enterococcus faecalis seed liquor: the enterococcus faecalis after activation is inoculated on enterococcus faecalis fluid nutrient medium, cultivates 24 hours for 37 ~ 39 DEG C; The preparation of bacillus subtilis seed liquor: the bacillus subtilis after activation is inoculated on bacillus subtilis fluid nutrient medium, cultivates 24 hours for 33 ~ 37 DEG C; The preparation of bacillus coagulans seed liquor: bacillus coagulans after activation is inoculated on bacillus coagulans fluid nutrient medium, cultivates 24 hours for 33 ~ 37 DEG C;
3. enterococcus faecalis seed liquor, bacillus subtilis seed liquor and bacillus coagulans seed liquor (1-10) by volume 2. step being prepared: (1-10): (1-10) mix, obtain seed mixture liquid, seed mixture liquid is inoculated in to the solidliquid mixture that 1. step obtains, temperature 35-40 DEG C of bottom fermentation 48-72 hour, dry, obtain feed addictive.
2. compound microorganism ferments is prepared the method for feed addictive as claimed in claim 1, it is characterized in that, the formula of described enterococcus faecalis fluid nutrient medium is: peptone 30 ~ 50g/L, glucose 30 ~ 40g/L, salt 5 ~ 10g/L, hepar siccatum 5 ~ 7g/L, pH6.5; The formula of bacillus subtilis fluid nutrient medium is: yeast soaks powder 3-5g/L, glucose 10-20g/L, magnesium sulfate 3-5g/L, pH6.0; The formula of bacillus coagulans fluid nutrient medium is: yeast soaks powder 3-5g/L, tryptone 2-5g/L, glucose 10-20g/L, calcium chloride 3-5g/L, pH6.0.
3. compound microorganism ferments is prepared the method for feed addictive as claimed in claim 1, it is characterized in that, described step 1. in sterilizing be 121 DEG C of sterilizings 30 minutes in autoclave.
4. compound microorganism ferments is prepared the method for feed addictive as claimed in claim 1, it is characterized in that, the preparation method of described Astragalus Root P.E is as follows: in astragalus membranaceus powder, add water, the quality that adds of water is 8 times of astragalus membranaceus powder quality, at 40-50 DEG C, flood 4 hours, then refluxing extraction 2 hours, obtains filter residue and filtrate; In filter residue, add 6 times to the water of filter residue quality, refluxing extraction 2 hours, filter, then merge the filtrate obtaining for twice, being concentrated into density is 1.15-1.25g/ml, add ethanol to obtain mixture, in mixture, ethanol mass content is 75%, leaves standstill 8 hours, pumps supernatant, to precipitate and be dried, be yellow its extract.
5. compound microorganism ferments is prepared the method for feed addictive as claimed in claim 1, it is characterized in that, described step 3. in the inoculum concentration of seed mixture liquid be step 1. in solidliquid mixture weight 10 ~ 20%.
6. compound microorganism ferments is prepared the method for feed addictive as claimed in claim 1, it is characterized in that, described step 2. middle enterococcus faecalis activation is at 38 DEG C, to cultivate and activate for 24 hours on slant medium, the formula that enterococcus faecalis activates slant medium used is: peptone 30 ~ 50g/L, glucose 30 ~ 40g/L, salt 5 ~ 10g/L, hepar siccatum 5 ~ 7g/L, agar 15-20g/L, pH6.5; Bacillus subtilis activation is at 35 DEG C, to cultivate and activate for 24 hours on slant medium, the formula that bacillus subtilis activates slant medium used is: yeast soaks powder 3-5g/L, glucose 10-20g/L, magnesium sulfate 3-5g/L, agar 15-20g/L, pH6.0; ; Bacillus coagulans activation is at 35 DEG C, to cultivate and activate for 24 hours on slant medium, and the formula of bacillus coagulans fluid nutrient medium is: yeast soaks powder 3-5g/L, tryptone 2-5g/L, glucose 10-20g/L, agar 15-20g/L, calcium chloride 3-5g/L, pH6.0.
7. the feed addictive of preparing according to the arbitrary described method of claim 1-6.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105941835A (en) * | 2016-06-30 | 2016-09-21 | 河南省岳氏精忠科技有限公司 | Method for preparing bacillus subtilis traditional Chinese medicine feed additive |
| CN106819472A (en) * | 2016-12-14 | 2017-06-13 | 康盛京构（北京）科技有限公司 | One boar biological feedstuff and preparation method thereof, purposes |
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| CN105941835A (en) * | 2016-06-30 | 2016-09-21 | 河南省岳氏精忠科技有限公司 | Method for preparing bacillus subtilis traditional Chinese medicine feed additive |
| CN106819472A (en) * | 2016-12-14 | 2017-06-13 | 康盛京构（北京）科技有限公司 | One boar biological feedstuff and preparation method thereof, purposes |
| CN107090485A (en) * | 2017-05-17 | 2017-08-25 | 内蒙古农业大学 | A kind of method and its special culture media and complex bacterial agent special for preparing active polysaccharide |
| CN107318979A (en) * | 2017-07-25 | 2017-11-07 | 东北农业大学 | A kind of spore type lactobacillus milk beverage and preparation method thereof |
| CN107865187A (en) * | 2017-11-16 | 2018-04-03 | 广西壮族自治区畜牧研究所 | A kind of purple napier grass feed and its preparation method and application |
| CN109170322A (en) * | 2018-09-17 | 2019-01-11 | 山西大禹生物工程股份有限公司 | Fermented tcm and preparation method thereof |
| CN109880754A (en) * | 2018-12-29 | 2019-06-14 | 广西康佳龙农牧集团有限公司 | A kind of bacillus liquid fermentation culture method |
| CN115025231A (en) * | 2022-06-15 | 2022-09-09 | 江西省药品检验检测研究院 | Dry inoculation method of traditional Chinese medicinal materials |
| CN115025231B (en) * | 2022-06-15 | 2024-03-19 | 江西省药品检验检测研究院 | Dry inoculation method of traditional Chinese medicinal materials |
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