CN103798503B - The ruminant Tiny ecosystem functional feed of composite zymocyte liquid and preparation method - Google Patents
The ruminant Tiny ecosystem functional feed of composite zymocyte liquid and preparation method Download PDFInfo
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Abstract
The ruminant Tiny ecosystem functional feed of composite zymocyte liquid of the present invention and preparation method belong to the technical field that biological bacteria is applied in feed, relate to the preferred LAg12-7 lactobacillus acidophilus of a strain, be used for, by after the fermentation of food industry discarded object, feedstuff and auxiliary material, being prepared into the method for ruminant Tiny ecosystem functional feed in particular to the composite fermentation microbial inoculum containing this bacterium.The product of LAg12-7 lactobacillus acidophilus is sour, acidproof and bacteriostasis property is good, and good compatibility can be had with bacillus subtilis and saccharomyces cerevisiae, pH value in fermented feed is made to be down to less than 4.0 rapidly, quick suppression harmful microorganism, and beneficial bacterium is had the advantage state all the time, fermented feed energy long term guarantee quality can be made.Ruminant Tiny ecosystem functional feed prepared by the present invention simple for process, production cost is low, feed conveniently, long shelf-life, and can improve the digestibility, good in economic efficiency of Dietary Fiber etc.
Description
Technical field
(the application is divisional application, original bill is Chinese patent application 201210399518.8) the invention belongs to the technical field that microorganism beneficial bacterium applies in feed, relate to the preferred lactobacillus acidophilus of a strain, in particular to containing this lactobacillus acidophilus composite fermentation microbial inoculum be used for using food industry discarded object as feedstuff and auxiliary material mixing after fermentation, be prepared into the method for ruminant Tiny ecosystem functional feed.
Background technology
Tiny ecosystem functional feed carries out the feed after fermentation process by multiple beneficial microorganism to relevant feedstuff, and the natural Tiny ecosystem system in microorganism wherein and metabolite and relevant animal body matches and nutrition complement.The Tiny ecosystem functional feed utilizing microbial fermentation technology to produce, for exploitation feed resource, improves efficiency of feed utilization, and regulate Tiny ecosystem in animal body average, substitute antibiotics, prevents and treats some disease, have positive effect.
Raw material for the production of fermented feed is very extensive, from the hydrocarbons such as early stage oil, methyl alcohol, ethanol food processing accessory substance, discarded object, agricultural crop straw etc. up till now.Wherein more to the research report utilizing grouts to produce fermentation protein feedstuff as raw material, object of such fermentation removes ANFs wherein and toxin, to improve the usage ratio of processing byproduct in mixed feed.And to food processing discarded object, as maize pulp-water, corn whitewashing fiber, sauce grain, manioc waste etc., because its Acidity of Aikalinity and salt content are not suitable for existing zymogenic fermentation work, and with zymophyte, maize pulp-water, corn whitewashing fiber, sauce grain, manioc waste etc. are not carried out fermentation manufacture feed.
Here is present maize pulp-water, corn whitewashing fiber, sauce grain, manioc waste Problems existing in feed applications:
Maize pulp-water refers to the suspension separated in production of corn starch process, and wherein BOD value is up to 1.5mg/g, is rich in organic substance, and general factory all drains into river, river, lake, causes serious pollution.According to surveying and determination, the moisture of maize pulp-water reaches 90.01%(and is permitted the sword autumn, 2002), concentrate maize pulp-water being condensed into moisture about 60% is called maize slurries, and its crude protein can reach content 10 ~ 15%.
Whitewashing zein fiber is the hybrid dry matter of maize pulp-water and maize peel, moisture about 10%, crude protein 22 ~ 24%, crude fibre about 11 ~ 13%.For utilize whitewashing zein fiber produce fermented feed there is not yet bibliographical information.
The poor slag that sauce grain is remaining after referring to and making soy sauce with soybean, rice, wheat bran etc., containing dry 41.47%, thick protein 12.72%, crude fat 5.31%, crude fibre 21.43%, NFE 0.62%, ash content 4.12%, calcium 0.08%, phosphorus 0.04%(Su Yu is virtuous, and 2001).Current most enterprise makes sauce grain powder and makes feed conversion after sauce grain being dried, but dry run energy consumption is high.
Manioc waste produces the waste material after starch or alcohol, the moisture more than 85.0% of fresh manioc waste, the moisture 10.0 ~ 12.0% of the manioc waste after oven dry with cassava, protein content about 4.0%, crude fiber content 12%.0, palatability is poor, and the value being directly used as feed is lower.
To utilizing food industry discarded object to produce fermented feed, there are reports both at home and abroad.Perhaps 2002 sword autumns reported and utilize geotrichum candidum fermented maize yellow seriflux to produce feed protein powder, but dry after needing press filtration, equipment investment is large, and this technology is still in laboratory research stage (Xu Jianqiu again, corn starch wastewater produces forage protein, grain and feed industries the 4th phase in 2002).Yan Qitao etc. (2000) utilize photosynthetic bacteria (PSB) fermented maize yellow seriflux to produce PSB bacterium mud (intercalation outstanding great waves etc., the application prospect of cornstarch yellow seriflux PSB method resource exploitation, Jilin Grain Advanced School's journal, 4th phase in 2000), this technology needs zymotic fluid to carry out the separation of bacterium liquid, make liquid qualified discharge, but be equally also that equipment investment is larger.
Summary of the invention
The technical problem to be solved in the present invention is to provide the preferred lactobacillus acidophilus of an a kind of strain for food industry discarded object and auxiliary material mixing after fermentation as Tiny ecosystem functional feed, and provides the method for this Tiny ecosystem functional feed of preparation.
The LAg12-7 bacterial strain that the present invention refers to, or LAg12-7 lactobacillus acidophilus is: LAg12-7
lactobacillusacidophiluslAg12-7.LAg12-7 strain isolation provided by the present invention is gastrointestinal tract mucous from yak, through be accredited as lactobacillus acidophilus (
lactobacillusacidophilus).LAg12-7
lactobacillusacidophiluslAg12-7 lactobacillus acidophilus preservation date: 2012 3 days on the 8th, depositary institution: China typical culture collection center, depositary institution address: China, Wuhan, Wuhan University (phone 027-68754052), survival detects August 7 2012 date: preserving number is: CCTCCNO:M2012305; This lactobacillus acidophilus has following characteristic:
(1) this bacterium is inoculated in MAS fluid nutrient medium, cultivates and within 22 hours, reach most Seedling height concentration under 37 DEG C of conditions, and OD600 value is shown in Fig. 1 for 2.86();
(2) this bacterium is inoculated in MRS fluid nutrient medium, and cultivate after 36 hours under 37 DEG C of conditions, the pH value of nutrient solution is shown in Fig. 2 lower than 3.50();
(3) this bacterium survival rate processed after 2 hours under pH2.0 condition is that 69.32%(is in table 1);
(4) this bacterium is after weight ratio is the cholate condition process 2 hours of 0.4% concentration, and the survival rate of bacterium is that 53.60%(is in table 2);
(5) this bacterium to the inhibition zone of Escherichia coli, staphylococcus aureus, salmonella and Shigella respectively: 12.14 ~ 17.30mm.
Ruminant Tiny ecosystem functional feed containing LAg12-7 lactobacillus acidophilus provided by the present invention at least has following a kind of function:
(1) feed intake of ruminant to roughage and the digestibility to dietary fiber is strengthened: 4.46% ~ 13.24%(is improved in table 6 to the feed intake of vinasse), crude fiber digestibility improves 9.73% ~ 19.02%;
(2) improve ruminant production performance and economic benefit: daily gain improves 7.89% ~ 8.77%, gross profit increases by 42.94% ~ 76.69%(in table 11);
(3) there is the function regulating ruminant tumor gastric microbial balance, increase the quantity of Rumen Cellulolytic Bacteria: (see table 9) is significantly improved to the quantity pole of yellow Ruminococcus etc., Ruminococcus albus, Lactobacillus acidophilus and bargen's streptococcus;
(4) protect ruminant body antioxidant system, improve non-specific immune function; strengthen premunition: the concentration of total protein, albumin and three kinds of each indexs of immunoglobulin (Ig) (IgA, IgG and IgM) in serum is significantly improved, the activity of TAC and superoxide dismutase is significantly improved (see table 10).
The present invention relates to bacterium, microbial inoculum, activation method, feed formula and feed making method technology contents as follows:
1, the acquisition of LAg12-7 lactobacillus acidophilus
Adopt the separation of conventional lactic acid bacteria, screening and identification method, 108 strains of lactic acid bacteria are obtained from the gastrointestinal tract mucous middle separation of yak, measured by physiological biochemical property, produce acid, acidproof salt tolerant experiment etc., therefrom filter out the lactobacillus acidophilus of a strain function admirable, called after LAg12-7 lactobacillus acidophilus, is made into LAg12-7 lactobacillus acidophilus freeze-drying powder through freeze drying.This bacterium is by China typical culture collection center (Wuhan University's preservation center) preservation, and it is numbered CCTCCM2012305, and the viable count of this bacterium freeze-dried powder can reach 20,000,000,000/gram.
2, the formula of composite fermentation microbial inoculum and the making of composite zymocyte liquid
Composite fermentation microbial inoculum component consists of by weight: LAg12-7 lactobacillus acidophilus freeze-drying powder 30.0 ~ 60.0%, bacillus subtilis freeze-dried powder 20.0 ~ 50.0%, saccharomyces cerevisiae freeze-dried powder 10.0 ~ 30.0%.
Above-mentioned LAg12-7 lactobacillus acidophilus is same as by China typical culture collection center (Wuhan University's preservation center) preservation, and it is numbered the LAg12-7 lactobacillus acidophilus of CCTCCM2012305, and its ice dry powder viable count is 20,000,000,000/gram; Bacillus subtilis freeze-dried powder purchased from magnificent occasion industrial (Chengdu) Science and Technology Ltd., its viable count is 20,000,000,000/gram; Saccharomyces cerevisiae freeze-dried powder purchased from Angel Yeast Co., Ltd, its viable count is 20,000,000,000/gram.
Before using composite fermentation microbial inoculum to ferment to substrate, composite fermentation microbial inoculum first need be activated and make conjunction zymocyte liquid.Prepare and close the method for zymocyte liquid and be: the brown sugar taking 5.0 ~ 15.0% by weight adds the clean water of 80.0 ~ 90.0%, after brown sugar dissolves, add composite fermentation microbial inoculum 0.3 ~ 1.0%, vitamin C 0.1 ~ 0.4%, urea 2.0 ~ 5.0% and potassium dihydrogen phosphate 0.1 ~ 0.5%, stir; Maintenance temperature and time is: be incubated 2 ~ 3 days under 32 DEG C of conditions, and between soak, every day stirs once, and solution ph can drop to less than 4.0, obtains the composite zymocyte liquid activated.
3, the compositing formula of ruminant Tiny ecosystem functional feed and preparation method
Need and feature of feeding according to ruminant animal nutrition, be first that the fermentation major ingredient of 75.0 ~ 85.0% mixes with the fermentation auxiliary material of 10.0 ~ 20.0% by weight ratio, the composite zymocyte liquid adding 2.0 ~ 6.0% is more fully mixed and made into fermentation raw material, then loaded by fermentation raw material in fermentation polybag, sealing and fermenting polybag makes fermentation raw material ferment in fermentation polybag.Fermentation condition was: environment temperature 15 ~ 30 DEG C of condition bottom fermentations 2 ~ 3 days, obtained finished product ruminant Tiny ecosystem functional feed; Or environment temperature 6 ~ 15 DEG C of condition bottom fermentations 5 ~ 7 days, obtain finished product ruminant Tiny ecosystem functional feed.
With the composition of top fermentation major ingredient and weight ratio be: take maize slurries 20.0 ~ 35.0% by weight, whitewashing zein fiber 25.0 ~ 45.0%, sauce grain 20.0 ~ 40.0% and manioc waste 5.0 ~ 15.0% directly mix.
Composition and the weight ratio of fermentation auxiliary material are: take wheat bran 50.0 ~ 65.0% by weight, beer yeast powder 20.0 ~ 35.0%, calcium carbonate 4.0 ~ 8.0%, sodium acid carbonate 4.0 ~ 8.0%, magnesium chloride 1.0 ~ 4.0% directly mix.
The composition of composite zymocyte liquid and weight ratio are: brown sugar 5.0 ~ 15.0%, clean water 80.0 ~ 90.0%, composite fermentation microbial inoculum 0.3 ~ 1.0%, vitamin C 0.1 ~ 0.4%, urea 2.0 ~ 5.0% and potassium dihydrogen phosphate 0.1 ~ 0.5%.
The preparation method of composite zymocyte liquid: the brown sugar taking 5.0 ~ 15.0% by weight adds the clean water of 80.0 ~ 90.0%, after brown sugar dissolves, add composite fermentation microbial inoculum 0.3 ~ 1.0%, vitamin C 0.1 ~ 0.4%, urea 2.0 ~ 5.0% and potassium dihydrogen phosphate 0.1 ~ 0.5%, stir; Maintenance temperature and time is: be incubated 2 ~ 3 days under 32 DEG C of conditions, and between soak, every day stirs once, and solution ph can drop to less than 4.0, obtains the composite zymocyte liquid activated.
In the present invention:
The water content of maize slurries is 58 ~ 60%;
The water content of whitewashing zein fiber is 10 ~ 12%;
The water content of sauce grain is 10 ~ 12%;
The water content of manioc waste is 10 ~ 12%.
Above-mentioned fermentation raw material: maize slurries are the concentrate of maize pulp-water simmer down to moisture 58 ~ 60%, purchased from state, Shandong biotechnology (Sichuan) Co., Ltd; Whitewashing zein fiber is the hybrid dry matter of maize pulp-water and maize peel, and moisture 10 ~ 12%, purchased from state, Shandong biotechnology (Sichuan) Co., Ltd; Sauce grain is that the recrement drying of producing soy sauce forms, and water content 10 ~ 12%, purchased from fermented glutinous rice limited company of Chengdu state; The water content of manioc waste is 10 ~ 12%, purchased from Jin Sheng source, Sichuan biochemical industry Co., Ltd; Beer yeast powder is in production beer process, and the recrement drying after beer fermentation liquid filters forms, and water content is 10%, purchased from golden prestige beer (Chengdu) Co., Ltd.
Advantage of the present invention: for domestic to food processing discarded object maize slurries at present, whitewashing zein fiber, the method of sauce grain and manioc waste exploitation feed product is single, equipment is complicated, the present situation that energy consumption is higher, integrated use microbial fermentation technology of the present invention and Animal nutrition principle, with food industry discarded object---maize slurries, corn whitewashing fiber, sauce grain and manioc waste are fermentation major ingredient, fermentation auxiliary material is added again according to the nutritional need feature of ruminant, add the composite zymocyte liquid after activation again (containing LAg12-7 lactobacillus acidophilus, bacillus subtilis and saccharomyces cerevisiae) after, solid state fermentation is carried out in direct loading fermentation polybag.The present invention has the following advantages:
1, excellent LAg12-7 lactobacillus acidophilus strain
The present invention, from 108 strains of lactic acid bacteria of the gastrointestinal tract mucous middle separation of yak, obtains the lactobacillus acidophilus of a strain function admirable through repeated screening, called after LAg12-7 lactobacillus acidophilus (number: CCTCCM2012305) by microbial preservation.This bacterium is inoculated in MRS fluid nutrient medium, and cultivate at 37 DEG C and within 22 hours, namely reach most Seedling height concentration, OD600 value is 2.86, cultivation 36 hours under 37 DEG C of conditions in MRS fluid nutrient medium, and the pH value of nutrient solution is namely lower than 3.50; Survival rate after pH2.0 process 2 hours is 69.32%; Survival rate after 0.7%% gallbladder salinity process 2 hours is 53.60%; All there is comparatively significantly inhibitory action to Escherichia coli, staphylococcus aureus, salmonella, Shigella, be made into LAg12-7 lactobacillus acidophilus freeze-drying powder through freeze drying, the viable count of freeze-dried powder is 20,000,000,000/gram.
Because the product of LAg12-7 lactobacillus acidophilus is sour, acidproof and bacteriostasis property is good, and good compatibility can be had with bacillus subtilis and saccharomyces cerevisiae, pH value in fermented feed is made to be down to less than 4.0 rapidly within 24 hours, quick suppression harmful microbe growth, and beneficial bacterium is had the advantage state all the time, fermented feed energy long term guarantee quality can be made.
2, the simple for process of ruminant Tiny ecosystem functional feed is prepared
This fermented feed changes general solid state fermentation pattern, pass through strain combination, the fermentation raw material prepared is put into fermentation polybag, carry out the aerobic three-dimensional energy-conservation fermentation combined with anaerobism: early stage ferments primarily of aerobic bacteria (saccharomycete), consume the oxygen in fermentation raw material, later stage ferments primarily of anaerobic bacteria (lactobacillus acidophilus), pH value in fermented feed is made to be reduced to less than 4.0 rapidly within 24 hours, quick suppression harmful microbe growth, bacillus subtilis breeding simultaneously produces the digestive ferments such as a large amount of cellulase, protease.Fermented through 7 ~ 20 days the ruminant Tiny ecosystem functional feed that gets product.Can the polybag of loading mass 40kg fermentation raw material namely be round, also be finished product packing, storage bag and shipping bags, need not pack again, need not be dry again, be directly transported to user.
3, ruminant Tiny ecosystem functional feed production cost is low
The present invention for primary raw material with food industry discarded object at a low price, adopts the energy-conservation zymotechnique that need not dry simultaneously, makes the production cost of feed low to gurry raw material.Due to by add suitable fermentation auxiliary material and and after fermenting containing the composite fermentation microbial inoculum of LAg12-7 lactobacillus acidophilus, become the ruminant Tiny ecosystem functional feed that there is better nutritivity and be worth.In ruminant fine fodder, add the Tiny ecosystem functional feed of the present invention of 20 ~ 60%, while raising ruminant production performance, ruminant feed cost about 4% can be reduced.
4, ruminant Tiny ecosystem functional feed is easy to use
The present invention is according to the nutritional need of ruminant and handling characteristics design, the nutritive index of the Tiny ecosystem functional feed after fermentation is made substantially to reach the nutritive index of ruminant fine fodder, user directly can substitute ruminant fine fodder with the weight ratio of 20 ~ 60% in use, need not change or adjust original ruminant Concentrate formula, easy to use.
5, ruminant Tiny ecosystem functional feed has several functions
Functional feed of the present invention contains the activated beneficial bacterium of multiple tool and has bioactive metabolite, has and strengthens the feed intake of ruminant to roughage and the digestibility to dietary fiber, improves production performance and economic benefit, quantity, the protection body antioxidant system of increase Rumen Cellulolytic Bacteria, improves non-specific immune function, strengthens the several functions such as premunition.
6, ruminant Tiny ecosystem functional feed long shelf-life
Ruminant Tiny ecosystem functional feed of the present invention is guaranteed the quality 70 ~ 90 days under environment temperature 30 DEG C ~ 40 DEG C conditions; Or guarantee the quality 150 ~ 180 days under environment temperature 0 ~ 30 DEG C of condition.And the existing feed containing lactobacillus acidophilus fermentation is guaranteed the quality 15 ~ 20 days under environment temperature 30 DEG C ~ 40 DEG C conditions; Or guarantee the quality 30 ~ 40 days under environment temperature 0 ~ 30 DEG C of condition.
Fig. 1 is the growth curve of LAg12-7 lactobacillus acidophilus;
Fig. 2 is the acid producing ability figure of LAg12-7 lactobacillus acidophilus.
Specific embodiment
The screening of embodiment 1, lactic acid bacteria and performance measurement
1.1, screen yak stomach and intestine lactic acid bacteria and obtain LAg12-7 lactobacillus acidophilus
MRS culture medium: peptone 10.00g, yeast extract 5.00g, beef extract 10.00g, glucose 20.00g, dipotassium hydrogen phosphate 2.00g, citric acid tri-amonia 1.00g, sodium acetate 5.00g, magnesium sulfate 0.80g, manganese sulfate 0.25g, Tween 80 1ml, 1000ml is settled to distilled water, pH6.2 ~ 6.4,115 DEG C of sterilizing 30min.
Adopt routine microbiological method, obtaining 108 strains of lactic acid bacteria from the gastrointestinal tract mucous middle separation of yak, by producing acid, acidproof salt tolerant experiment etc., therefrom filtering out the bacterial strain of a strain function admirable.This bacterial strain to be identified and 16SrDNA Molecular Identification determines that this bacterial strain is lactobacillus acidophilus, called after lactobacillus acidophilus LAg12-7 through Physiology and biochemistry, and bacterial strain is made into freeze-dried powder through freeze drying, the viable count of freeze-dried powder is 20,000,000,000/gram.This bacterium is by China typical culture collection center (Wuhan University's preservation center) preservation, and it is numbered CCTCCM2012305.
1.2, LAg12-7 lactobacillus acidophilus growth test
Lactobacillus acidophilus LAg12-7 is proceeded to 37 DEG C of activation 15 ~ 18 in MRS fluid nutrient medium little.The bacterial strain activated is proceeded to Anaerobic culturel in MRS fluid nutrient medium with the inoculum concentration of 2% (V/V), sampling and measuring OD600 value at regular intervals, using nonvaccinated culture medium as blank.Result can see above LAg12-7 lactobacillus acidophilus breeding in 20 hours rapidly, and OD600 value continues to rise; From 20 ~ 48 hours, cell density tended to balance substantially, and OD600 value stabilization, about 2.86, is in and grows stationary phase, see Fig. 1.
1.3, LAg12-7 lactobacillus acidophilus produces acid test
After being activated by LAg12-7 lactobacillus acidophilus, being inoculated into MRS fluid nutrient medium, cultivating under 37 DEG C of conditions, at regular intervals sampling and measuring pH value, is initial pH with the pH of nonvaccinated MRS fluid nutrient medium.The results are shown in Figure 2, visible cultivation is after 36 hours, and the pH value of LAg12-7 lactobacillus acidophilus nutrient solution is lower than 3.50.
After the activation of LAg12-7 lactobacillus acidophilus, inoculation MRS fluid nutrient medium, cultivate under 37 DEG C of conditions, measuring zymotic fluid pH after cultivating certain hour, is initial pH with the pH of nonvaccinated MRS fluid nutrient medium.Result shows, after this Spawn incubation, pH declines fast, and after cultivating 36h, pH downward trend eases up, and cultivation reaches 3.45. to pH during 60h and cultivates after 36 hours as seen, and the pH value of LAg12-7 lactobacillus acidophilus nutrient solution, lower than 3.50, is shown in Fig. 2.
1.4, LAg12-7 lactobacillus acidophilus acid resistance test
Strain inoculation is activated 2 times on MRS fluid nutrient medium, with 2%(V/V) inoculum concentration be inoculated in MRS fluid nutrient medium, 24h hour is cultivated under 37 DEG C of conditions, centrifugal 10 minutes of 8000rpm/min, collect thalline, by thalline pbs(pH6.0) buffer solution 1 time, Eddy diffusion is in pbs(pH6.0) in buffer solution.Suspension is divided into 5 points, again collected by centrifugation thalline.The thalline obtained is suspended in respectively pH2.0,3.0,4.0, in the pbs buffer solution of 5.0 and 6.0, the pbs bacterium liquid of the 5 parts of different pH obtained, at room temperature places 2 hours, carries out count plate, with the viable count under pH6.0 condition for 100%, calculate its survival rate.
LAg12-7 lactobacillus acidophilus acid resistance test the results are shown in Table 1, shows that LAg12-7 lactobacillus acidophilus survival rate of placing after 2 hours under pH2.0 condition is 69.32%.
Table 1LAg12-7 lactobacillus acidophilus survival rate at various ph values (unit: %)
1.5, LAg12-7 lactobacillus acidophilus bile tolerance test
Cholate addition establishes 4 gradients: 0.1%, 0.3%, 0.5% and 0.7%, not add the culture medium of cholate for contrast.Strain inoculation activated 2 times on MRS fluid nutrient medium, be inoculated in treatment samples and control sample with 2% inoculum concentration, under 37 DEG C of conditions, quiescent culture carried out count plate after 2 hours.
LAg12-7 lactobacillus acidophilus bile tolerance result of the test is in table 2, and showing that LAg12-7 lactobacillus acidophilus cultivates the survival rate after 2 hours under 0.7% cholate condition is 53.60%.
The survival rate of table 2 lactic acid bacteria under different salinity (unit: %)
The survival rate of table 2 lactic acid bacteria under different salinity (unit: %)
1.6, LAg12-7 lactobacillus acidophilus bacteriostatic test
Select Escherichia coli, staphylococcus aureus, salmonella, Shigella as indicator bacteria.Bacteriostatic activity analytical method: under the incident light condition of 600 λ, the OD value of indicator bacteria bacteria suspension is adjusted to 0.10, be poured into 15mLLB solid medium, after culture medium solidifying, place the Oxford cup that diameter is 7mm, add the fermented supernatant fluid of lactobacillus acidophilus LAg12-7, at room temperature spread 30min, under 37 DEG C of conditions, cultivate 16-18 hour, observe inhibition zone.
The method of the composite fermentation microbial inoculum of embodiment 2, preparation ruminant Tiny ecosystem functional feed
The composition of this composite fermentation microbial inoculum and weight ratio are: LAg12-7 lactobacillus acidophilus freeze-drying powder 30.0%, bacillus subtilis freeze-dried powder 50.0%, saccharomyces cerevisiae freeze-dried powder 20.0%.LAg12-7 lactobacillus acidophilus freeze-drying powder, bacillus subtilis freeze-dried powder and saccharomyces cerevisiae freeze-dried powder are mixed under normal temperature condition the composite fermentation microbial inoculum just making ruminant Tiny ecosystem functional feed.
The method of the composite fermentation microbial inoculum of embodiment 3, preparation ruminant Tiny ecosystem functional feed
The composition of this composite fermentation microbial inoculum and weight ratio are: LAg12-7 lactobacillus acidophilus freeze-drying powder 60.0%, bacillus subtilis freeze-dried powder 20.0%, saccharomyces cerevisiae freeze-dried powder 20.0%.LAg12-7 lactobacillus acidophilus freeze-drying powder, bacillus subtilis freeze-dried powder and saccharomyces cerevisiae freeze-dried powder are mixed under normal temperature condition the composite fermentation microbial inoculum just making ruminant Tiny ecosystem functional feed.
The method of the composite fermentation microbial inoculum of embodiment 4, preparation ruminant Tiny ecosystem functional feed
The composition of this composite fermentation microbial inoculum and weight ratio are: LAg12-7 lactobacillus acidophilus freeze-drying powder 45.0%, bacillus subtilis freeze-dried powder 45.0%, saccharomyces cerevisiae freeze-dried powder 10.0%.LAg12-7 lactobacillus acidophilus freeze-drying powder, bacillus subtilis freeze-dried powder and saccharomyces cerevisiae freeze-dried powder are mixed under normal temperature condition the composite fermentation microbial inoculum just making ruminant Tiny ecosystem functional feed.
The method of the composite fermentation microbial inoculum of embodiment 5, preparation ruminant Tiny ecosystem functional feed
The composition of this composite fermentation microbial inoculum and weight ratio are: LAg12-7 lactobacillus acidophilus freeze-drying powder 40.0%, bacillus subtilis freeze-dried powder 30.0%, saccharomyces cerevisiae freeze-dried powder 30.0%.LAg12-7 lactobacillus acidophilus freeze-drying powder, bacillus subtilis freeze-dried powder and saccharomyces cerevisiae freeze-dried powder are mixed under normal temperature condition the composite fermentation microbial inoculum just making ruminant Tiny ecosystem functional feed.
Embodiment 6, composite fermentation microbial inoculum is activated into the method for composite zymocyte liquid
The composition of composite zymocyte liquid and weight ratio are: brown sugar 5.0%, composite fermentation microbial inoculum 1.0%, vitamin C 0.4%, urea 3.1%, potassium dihydrogen phosphate 0.5% and clean water 90.0%.
Step by composite fermentation microbial inoculum activates: take brown sugar by above-mentioned formula and add clean water, after brown sugar dissolves, add composite fermentation microbial inoculum, vitamin C, urea and potassium dihydrogen phosphate, stir; Maintenance temperature and time is: be incubated 2 ~ 3 days under 32 DEG C of conditions, and between soak, every day stirs once, and solution ph can drop to less than 4.0, obtains the composite zymocyte liquid activated.
Embodiment 7, composite fermentation microbial inoculum is activated into the method for composite zymocyte liquid
The composition of composite zymocyte liquid and weight ratio are: brown sugar 15.0%, composite fermentation microbial inoculum 0.3%, vitamin C 0.1%, urea 2.0%, potassium dihydrogen phosphate 0.1% and clean water 82.5%.The step that composite fermentation microbial inoculum activates is same as embodiment 6.
Embodiment 8, composite fermentation microbial inoculum is activated into the method for composite zymocyte liquid
The composition of composite zymocyte liquid and weight ratio are: brown sugar 8.70%, composite fermentation microbial inoculum 0.52%, vitamin C 0.17%, urea 5.00%, potassium dihydrogen phosphate 0.17% and clean water 85.43%.The step that composite fermentation microbial inoculum activates is same as embodiment 6.
Embodiment 9, composite fermentation microbial inoculum is activated into the method for composite zymocyte liquid
The composition of composite zymocyte liquid and weight ratio are: brown sugar 13.1%, composite fermentation microbial inoculum 1.0%, vitamin C 0.4%, urea 5.0%, potassium dihydrogen phosphate 0.5% and clean water 80.0%.The step that composite fermentation microbial inoculum activates is same as embodiment 6.
The preparation method of embodiment 10, ruminant Tiny ecosystem functional feed
In ruminant Tiny ecosystem functional feed, the formula of each raw material is as follows:
Composition and the weight ratio of fermentation major ingredient are: maize slurries 20.0%, whitewashing zein fiber 45.0%, sauce grain 20.0% and manioc waste 15.0%;
The fermentation composition of auxiliary material and weight ratio are: wheat bran 50.0%, beer yeast powder 35.0%, calcium carbonate 8.0%, 6.0%, magnesium chloride 1.0%;
The composition of composite zymocyte liquid and weight ratio are: brown sugar 5.0%, composite fermentation microbial inoculum 1.0%, vitamin C 0.4%, urea 3.1%, potassium dihydrogen phosphate 0.5% and clean water 90.0%;
The raw material composition of ruminant Tiny ecosystem functional feed and weight ratio are: fermentation major ingredient 75.0%, fermentation auxiliary material 20.0%, composite zymocyte liquid 5.0%;
Wherein the water content of part material is: the water content of maize slurries is 58 ~ 60%; The water content of whitewashing zein fiber is 10 ~ 12%; The water content of sauce grain is 10 ~ 12%; The water content of manioc waste is 10 ~ 12%.
The step making ruminant Tiny ecosystem functional feed of above-mentioned raw materials is as follows:
(1) fermentation major ingredient is made: take maize slurries, whitewashing zein fiber, sauce grain and manioc waste by weight and directly mix;
(2) fermentation auxiliary material is made: take wheat bran by weight, beer yeast powder, calcium carbonate, sodium acid carbonate, magnesium chloride directly mix;
(3) composite zymocyte liquid is made: take brown sugar by weight and add clean water 80.0 ~ 90.0%, after brown sugar dissolves, add composite fermentation microbial inoculum, vitamin C, urea and potassium dihydrogen phosphate, stir; Maintenance temperature and time is: be incubated 2 ~ 3 days under 2 DEG C of conditions, and between soak, every day stirs once, and solution ph can drop to less than 4.0, obtains the composite zymocyte liquid activated;
(4) make functional feed: first with weight ratio be fermentation major ingredient with ferment auxiliary material mix, add composite zymocyte liquid again and be fully mixed and made into fermentation raw material, then loaded by fermentation raw material in fermentation polybag, sealing and fermenting polybag makes fermentation raw material ferment in fermentation polybag; Fermentation condition was: environment temperature 15 ~ 30 DEG C of condition bottom fermentations 2 ~ 3 days, obtained finished product ruminant Tiny ecosystem functional feed; Or environment temperature 6 ~ 15 DEG C of condition bottom fermentations 5 ~ 7 days, obtain finished product ruminant Tiny ecosystem functional feed.
The preparation method of embodiment 11, ruminant Tiny ecosystem functional feed
In ruminant Tiny ecosystem functional feed, the formula of each raw material is as follows:
Composition and the weight ratio of fermentation major ingredient are: maize slurries 35.0%, whitewashing zein fiber 25.0%, sauce grain 35.0% and manioc waste 5.0%;
Composition and the weight ratio of fermentation auxiliary material are: wheat bran 65.0%, beer yeast powder 20.0%, calcium carbonate 4.0%, sodium acid carbonate 8.0%, magnesium chloride 3.0%;
The composition of composite zymocyte liquid and weight ratio are: brown sugar 15.0%, composite fermentation microbial inoculum 0.3%, vitamin C 0.1%, urea 2.0%, potassium dihydrogen phosphate 0.1% and clean water 82.5%;
The raw material composition of ruminant Tiny ecosystem functional feed and weight ratio are: fermentation major ingredient 85.0%, fermentation auxiliary material 11.0%, composite zymocyte liquid 4.0%;
Wherein the water content of part material is same as embodiment 10.
The step making ruminant Tiny ecosystem functional feed of above-mentioned raw materials is same as embodiment 10.
The preparation method of embodiment 12, ruminant Tiny ecosystem functional feed
In ruminant Tiny ecosystem functional feed, the formula of each raw material is as follows:
Composition and the weight ratio of fermentation major ingredient are: maize slurries 25.0%, whitewashing zein fiber 30.0%, sauce grain 40.0% and manioc waste 5.0%;
Composition and the weight ratio of fermentation auxiliary material are: wheat bran 60.0%, beer yeast powder 26.0%, calcium carbonate 6.0%, sodium acid carbonate 4.0%, magnesium chloride 4.0%;
The composition of composite zymocyte liquid and weight ratio are: brown sugar 8.70%, composite fermentation microbial inoculum 0.52%, vitamin C 0.17%, urea 5.00%, potassium dihydrogen phosphate 0.17% and clean water 85.43%.The step that composite fermentation microbial inoculum activates is same as embodiment 6;
The raw material composition of ruminant Tiny ecosystem functional feed and weight ratio are: fermentation major ingredient 88.0%, fermentation auxiliary material 10.0%, composite zymocyte liquid 2.0%;
Wherein the water content of part material is same as embodiment 10.
The step making ruminant Tiny ecosystem functional feed of above-mentioned raw materials is same as embodiment 10.
The preparation method of embodiment 13, ruminant Tiny ecosystem functional feed
In ruminant Tiny ecosystem functional feed, the formula of each raw material is as follows:
Composition and the weight ratio of fermentation major ingredient are: maize slurries 25.0%, whitewashing zein fiber 30.0%, sauce grain 40.0% and manioc waste 5.0%;
Composition and the weight ratio of fermentation auxiliary material are: wheat bran 60.0%, beer yeast powder 26.0%, calcium carbonate 6.0%, sodium acid carbonate 4.0%, magnesium chloride 4.0%;
The composition of composite zymocyte liquid and weight ratio are: brown sugar 13.1%, composite fermentation microbial inoculum 1.0%, vitamin C 0.4%, urea 5.00%, potassium dihydrogen phosphate 0.5% and clean water 8.0%.The step that composite fermentation microbial inoculum activates is same as embodiment 6;
The raw material composition of ruminant Tiny ecosystem functional feed and weight ratio are: fermentation major ingredient 79%, fermentation auxiliary material 15.0%, composite zymocyte liquid 6.0%;
Wherein the water content of part material is same as embodiment 10.
The step making ruminant Tiny ecosystem functional feed of above-mentioned raw materials is same as embodiment 10.
Embodiment 14, beef cattle Tiny ecosystem functional feed feeding experiment
The present embodiment adds the ruminant Tiny ecosystem functional feed of the present invention of 25% or 40% in fattening beef cattle fine fodder, the feed intake 4.46% ~ 13.24% of beef cattle to coarse fodder can be increased, improve daily gain 7.89% ~ 8.77%, improve crude fiber digestibility 9.73% ~ 19.02%, increase the quantity 1.28% ~ 74.48% of Rumen Cellulolytic Bacteria and lactic acid bacteria, improve beef cattle non-specific immune function, thus (gross profit) 42.94% ~ 76.69% of increasing economic efficiency.From this sample result, be good with the effect of the Tiny ecosystem functional feed adding 40% in basic fine fodder.Here is concrete feeding experiment and the mensuration to indexs such as the test production performance of beef cattle, nutrient digestion, rumen microorganism, Physiology and biochemistry and immunity:
1, testing program
1.1, beef cattle Tiny ecosystem functional feed: with the ruminant Tiny ecosystem functional feed containing lactobacillus acidophilus of the present invention.
1.2, experimental design
Test is carried out in certain large-scale steer ration field.The consistent west of body weight 373.40 ± 42.53kg, body condition hair color is selected to mix ox 90, adopt 5 × 3 RANDOMIZED BLOCK DESIGN, according to body weight phase approximately principle, 90 oxen are divided into 5 district's groups, each district group 18,18 oxen of same district group are divided into randomly individual process (control group, test I group and II group), each process 5 repetition, each repetition 6 oxen; Control group fed basis fine fodder, test I group and II group respectively with Tiny ecosystem functional feed with 25% and 40% ratio substitute basic fine fodder, respectively organize fine fodder (4kg) and the corn stalk pellets (2.7kg) of identical weight of feeding every every day, vinasse free choice feeding, fasten and feed, freely drink water; Test period: raise 10 days phases in advance, is just trying 48 days phases; Every day records feed intake, ox morning every of on-test and end is weighed on an empty stomach, during off-test, each repetition is selected to butcher close to 3 oxen of this repetition average weight; get longissimus dorsi muscle and measure pork quality traits; get the quantity that rumen fluid measures rumen microorganism, jugular vein blood collection measures serum biochemistry and immune indexes.
1.3, feed formula and nutritional labeling
Control group (basic fine fodder) is fine fodder original in field, and the test I group ruminant Tiny ecosystem functional feed containing lactobacillus acidophilus of the present invention substitutes 25% of basic fine fodder weight, and test II group is for 40% of basic fine fodder weight.The weight ratio formula of each processed group fine fodder is in table 4.
The composition of each processed group fine fodder of table 4 and nutritional labeling weight ratio (%)
| Material name | Control group (basic fine fodder) | Test I group (25%) | Test II group (40%) |
| Corn | 69.00 | 51.75 | 41.40 |
| Wheat bran | 10.00 | 7.50 | 6.00 |
| Rice spermoderm | 8.00 | 6.00 | 4.80 |
| Cottonseed Meal | 3.00 | 2.25 | 1.80 |
| Rapeseed dregs | 3.00 | 2.25 | 1.80 |
| Sodium bicarbonate | 1.50 | 1.13 | 0.90 |
| Salt | 1.00 | 0.75 | 0.60 |
| Sodium sulphate | 0.50 | 0.38 | 0.30 |
| Premix | 4.00 | 3.00 | 2.40 |
| The Tiny ecosystem functional feed of embodiment 10 | - | 25.00 | 40.00 |
| Add up to | 100 | 100 | 100 |
1.4, testing index and method thereof
1.4.1, production performance
Every beef cattle to be fed the corn stalk pellets feed of 4kg fine fodder and 1.7kg every day, and alcohol free choice feeding, records the vinasse feed intake of each repetition every day, calculates every beef cattle full phase average feed intake; On-test and at the end of, every beef cattle morning weighs on an empty stomach, calculates the average daily gain of every beef cattle.
1.4.2, nutrient digestibility
First three sky of off-test gathers Feed Sample and excrement sample, adopts 4NHCl acid insoluble ash (AIA) method to measure protein and coarse-fibred digestibility.The computing formula of AIA method digestibility is as follows:
1.4.3, the mensuration of rumen microorganism quantity
Beef cattle gets rumen fluid after butchering, and adopt RT-PCR absolute quantification method to measure the quantity of rumen microorganism, the four kinds of microorganisms measured and 16SrDNA fragment primer sequence thereof are as table 5.
Table 5 increases the 16SrDNA fragment primer sequence of yellow Ruminococcus, Ruminococcus albus, lactic acid bacteria and bargen's streptococcus
1.4.3, the mensuration of Biochemical Indices In Serum, immune indexes and Antioxidant Indexes
Jugular vein blood 20ml, centrifugal 15 minutes of 3000prm after left at room temperature half an hour, obtained serum-20 DEG C of refrigerators are recommended with guarantee stand-by.Total serum protein (BCA method), albumin (Bromocresol green), glutamic-pyruvic transaminase/glutamic-oxalacetic transaminease (reitman-frankel method), immunoglobulin (Ig) (IgA, IgG and IgM), TAC, total number born and concentration of malondialdehyde all adopt kit measurement.Used kit all builds up Bioengineering Research Institute purchased from Nanjing, and all operations carries out in strict accordance with kit operation instruction.
2, the feeding experiment result of functional feed
2.1, ruminant Tiny ecosystem functional feed is on the impact of beef cattle production performance
2.1.1, ruminant Tiny ecosystem functional feed is on the impact of beef cattle feed intake
Each process every ox to be fed fine fodder 4kg/ head every day. sky, corn stalk pellets 2.7kg/ head, vinasse free choice feeding.Each group of feed intake the results are shown in Table 6.As shown in Table 6, test I group and II group to be searched for food vinasse 0.89kg and 0.30kg than control group respectively, respectively raising 13.24% and 4.46%, but difference remarkable (P > 0.05).
Table 6 Tiny ecosystem functional feed is on the impact of beef cattle feed intake
2.1.2, daily gain
Daily gain result shows, duration of test, test I group and II group every beef cattle weightening finish 4.59kg and 4.91kg more than many control groups respectively, and daily gain you can well imagine high 7.89% and 8.77% than contrast component, but each group difference is not significantly (P > 0.05).
Tiny ecosystem functional feed is on the impact of beef cattle daily gain
2.2, ruminant Tiny ecosystem functional feed is on the impact on cattle diet nutrient digestibility
The nutrient digestibility result of variations of each processed group shows, adds Tiny ecosystem functional feed in fine fodder, has the trend increasing crude protein digestibility, but difference is not significantly (P > 0.05); Pole significantly improves coarse-fibred digestibility (P < 0.01).
Tiny ecosystem functional feed is on the impact of cattle diet nutrient digestibility
2.3, on the impact of rumen microorganism
Yellow Ruminococcus and Ruminococcus albus are cellulose-decomposing bacterias main in cud, and Lactobacillus acidophilus and bargen's streptococcus are Lactic Acid Producings main in cud.As can be seen from Table 9, Tiny ecosystem functional feed increases Rumen Cellulolytic Bacteria (yellow Ruminococcus and Ruminococcus albus) and the quantity of Lactic Acid Producing (Lactobacillus acidophilus and bargen's streptococcus), and has the trend of increase with the increase of Tiny ecosystem functional feed addition.
Table 9 Tiny ecosystem functional feed is on the impact (unit: × 10 of rumen microorganism quantity
8copy number/μ l, %)
2.4, on the impact of beef cattle Serum Growth, immunity and Antioxidant Indexes
Tiny ecosystem functional feed on the impact of beef cattle Serum Growth, immunity and Antioxidant Indexes in table 10.
In Biochemical Indices In Serum, the total serum protein of test group, albumin content are significantly higher than control group (P < 0.01), and the serum glutamic oxalacetic transaminase (P < 0.05) of test group and glutamic-pyruvic transaminase (P < 0.01) content are significantly or extremely remarkable in control group; But the Biochemical Indices In Serum difference between two test group is not all significantly (P > 0.05).
Immunoglobulin IgA in serum, IgG and IgM, and TAC and total number born content pole are significantly higher than control group (P < 0.01), and be increase trend with the increase of Tiny ecosystem functional feed addition; And test group mda content is extremely remarkable in control group (P < 0.01).
Table 10 Tiny ecosystem functional feed is on the impact of beef cattle serum biochemistry, immunity and Antioxidant Indexes
In serum, total protein is made up of albumin and globulin.Albumin is not only the main body forming plasma colloid osmotic pressure, is also the transport agent of water-soluble lower material in blood.Glutamic-oxalacetic transaminease and glutamic-pyruvic transaminase are two important indicators of reflection liver and cardiac function, and their hyperactivities just mean that liver and heart can be detrimental to evil.Immunoglobulin (Ig) is an important component part of body immune system.Superoxide dismutase can remove ultra-oxygen anion free radical, and Cell protection, from damage, plays vital effect to the oxidative and anti-oxidative balance of body.MDA is the end-product of the MDA that produces of peroxidatic reaction of lipid that oxygen radical causes at body intracellular metabolite, indirectly can reflect the degree that the situation of radical metabolism in body, body tissue cell are attacked by free radical and level of lipid peroxidation.
This research shows, two test group all can improve the concentration of total protein in serum, albumin and immunoglobulin (Ig) (IgA, IgG and IgM); Reduce glutamic-oxalacetic transaminease, gpt activity and concentration of malondialdehyde; And strengthen TAC, improve the activity of total number born.Illustrate that Tiny ecosystem functional feed can protect beef cattle body antioxidant system, improve beef cattle non-specific immune function.
2.5, Economic Benefit Accounting and analysis
The fine fodder cost of control group, test I group and II group is respectively 2.29 yuan/kg, 2.31 yuan/kg and 2.26 yuan/kg, the cost of corn stalk pellets in 0.98 yuan, the cost of vinasse is with 0.80 yuan, according to the feed intake of table 6, the feed cost calculating each processed group every every day is respectively 17.18 yuan, 17.97 yuan and 17.58 yuan.Cattle on the hoof commercial value is in 16.5 yuan of kg, and the income calculating each processed group every every day is respectively 18.81 yuan, 20.30 yuan and 20.46 yuan.Than control group 1.63 yuan/day. the gross profit of head increases income 0.70 yuan and 1.25 yuan (table 11) more.
Table 11 Economic Benefit Accounting table
Claims (2)
1. the ruminant Tiny ecosystem functional feed of composite zymocyte liquid, is characterized in that: raw material composition and weight ratio are: fermentation major ingredient 75.0 ~ 85.0%, fermentation auxiliary material 10.0 ~ 20.0%, composite zymocyte liquid 2.0 ~ 6.0%;
Composition and the weight ratio of fermentation major ingredient are: maize slurries 20.0 ~ 35.0%, whitewashing zein fiber 25.0 ~ 45.0%, sauce grain 20.0 ~ 40.0% and manioc waste 5.0 ~ 15.0%, wherein: the water content of maize slurries is 58.0 ~ 60.0%, the water content of whitewashing zein fiber is 10.0 ~ 12.0%, the water content of sauce grain is 10.0 ~ 12.0%, and the water content of manioc waste is 10.0 ~ 12.0%;
Composition and the weight ratio of fermentation auxiliary material are: wheat bran 50.0 ~ 65.0%, beer yeast powder 20.0 ~ 35.0%, calcium carbonate 4.0 ~ 8.0%, sodium acid carbonate 4.0 ~ 8.0%, magnesium chloride 1.0 ~ 4.0%;
The composition of composite zymocyte liquid and weight ratio are: brown sugar 5.0 ~ 15.0%, clean water 80.0 ~ 90.0%, composite fermentation microbial inoculum 0.3 ~ 1.0%, vitamin C 0.1 ~ 0.4%, urea 2.0 ~ 5.0% and potassium dihydrogen phosphate 0.1 ~ 0.5%;
In above-mentioned Tiny ecosystem functional feed, the composition of composite fermentation microbial inoculum and weight ratio are: LAg12-7 lactobacillus acidophilus freeze-drying powder 30.0 ~ 60.0%, bacillus subtilis freeze-dried powder 20.0 ~ 50.0%, saccharomyces cerevisiae freeze-dried powder 10.0 ~ 30.0%; Wherein: LAg12-7 lactobacillus acidophilus is by China typical culture collection center (Wuhan University's preservation center) preservation, and its deposit number is CCTCCM2012305.
2. the preparation method of ruminant Tiny ecosystem functional feed according to claim 1, is characterized in that this feed making method is as follows:
(1) fermentation major ingredient is made: take maize slurries 20.0 ~ 35.0% by weight, whitewashing zein fiber 25.0 ~ 45.0%, sauce grain 20.0 ~ 40.0% and manioc waste 5.0 ~ 15.0% directly mix;
(2) fermentation auxiliary material is made: take wheat bran 50.0 ~ 65.0% by weight, beer yeast powder 20.0 ~ 35.0%, calcium carbonate 4.0 ~ 8.0%, sodium acid carbonate 4.0 ~ 8.0%, magnesium chloride 1.0 ~ 4.0% directly mix;
(3) composite zymocyte liquid is made: the brown sugar taking 5.0 ~ 15.0% by weight adds the clean water of 80.0 ~ 90.0%, after brown sugar dissolves, add composite fermentation microbial inoculum 0.3 ~ 1.0%, vitamin C 0.1 ~ 0.4%, urea 2.0 ~ 5.0% and potassium dihydrogen phosphate 0.1 ~ 0.5%, stir; Maintenance temperature and time is: be incubated 2 ~ 3 days under 32 DEG C of conditions, and between soak, every day stirs once, and solution ph can drop to less than 4.0, obtains the composite zymocyte liquid activated;
(4) functional feed is made: be first that the fermentation major ingredient of 75.0 ~ 85.0% mixes with the fermentation auxiliary material of 10.0 ~ 20.0% by weight ratio, the composite zymocyte liquid adding 2.0 ~ 6.0% is more fully mixed and made into fermentation raw material, then loaded by fermentation raw material in fermentation polybag, sealing and fermenting polybag makes fermentation raw material ferment in fermentation polybag; Fermentation condition was: environment temperature 15 ~ 30 DEG C of condition bottom fermentations 2 ~ 3 days, obtained finished product ruminant Tiny ecosystem functional feed; Or environment temperature 6 ~ 15 DEG C of condition bottom fermentations 5 ~ 7 days, obtain finished product ruminant Tiny ecosystem functional feed.
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| CN107751568A (en) * | 2017-10-31 | 2018-03-06 | 南京工业大学大丰海洋产业研究院 | A kind of method that Spartina alterniflora's biofermentation prepares Livestock feed |
| CN108185112A (en) * | 2017-12-27 | 2018-06-22 | 河南飞天农业开发股份有限公司 | A kind of production technology of plant protein fodder |
| CN108634111A (en) * | 2018-04-12 | 2018-10-12 | 西南民族大学 | The special preparation method for licking brick of Adult Yak warm season |
| CN109007308A (en) * | 2018-08-10 | 2018-12-18 | 山东泽康农业科技有限公司 | A kind of micro-zoology composite feed and preparation method thereof |
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| CN110934223A (en) * | 2019-12-31 | 2020-03-31 | 内蒙古阜丰生物科技有限公司 | Process for preparing protein feed by fermenting corn bran solid microorganisms |
| CN111202177B (en) * | 2020-01-17 | 2023-05-16 | 安徽省农业科学院农产品加工研究所 | Fermented yellow serofluid bioactive feed for healthy pig breeding and preparation method thereof |
| CN111202172B (en) * | 2020-01-17 | 2023-04-07 | 安徽省农业科学院农产品加工研究所 | Pasture grass silage bacterial agent fermented by soybean yellow serofluid and application of pasture grass silage bacterial agent |
| CN112608854A (en) * | 2021-02-03 | 2021-04-06 | 邵阳学院 | Preparation method of yeast with high acid resistance |
| CN113439798A (en) * | 2021-05-28 | 2021-09-28 | 山西农业大学 | Enzyme and microecological preparation compound agent and preparation method thereof |
| CN113475639A (en) * | 2021-07-09 | 2021-10-08 | 山西农业大学 | Application of enzyme and microecological preparation compound composition in weaned pig feed and method thereof |
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