CN103798503A - Ruminant micro-ecological functional feed prepared through compound fermentative bacteria solution and preparation method of feed - Google Patents
Ruminant micro-ecological functional feed prepared through compound fermentative bacteria solution and preparation method of feed Download PDFInfo
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Abstract
The invention relates to a ruminant micro-ecological functional feed prepared through a compound fermentative bacteria solution and a preparation method of the feed, and belongs to the technical field of application of biological bacteria in feeds. The invention relates to a preferred LAg12-7 lactobacillus acidophilus, and particularly relates to a method for preparing the ruminant micro-ecological functional feed after food industry wastes, feed raw materials and auxiliary materials are fermented by using the compound fermentative bacteria solution which contains the LAg12-7 lactobacillus acidophilus. The preferred LAg12-7 lactobacillus acidophilus disclosed by the invention has a good acid-producing property, an acid-resisting property and a bacteria-inhibiting property, achieves the very good compatibility with bacillus subtilis and saccharomyces cerevisiae, fast reduces the pH value of a fermentive feed below 4.0, fast inhibits harmful microbes and enables that the quality of the fermentive feed can be guaranteed for a long time under the condition that beneficial bacteria are in a preponderant state all the time. The ruminant micro-ecological functional feed prepared by adopting the method disclosed by the invention has the advantages of simplicity and easiness for operation of a process, low production cost, convenience for feeding, long guarantee period, enhancement of the digestibility of daily ration cellulose and the like, and good economic benefits.
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 is applied in feed, relate to the preferred lactobacillus acidophilus of a strain, be particularly related to the composite fermentation microbial inoculum that contains this lactobacillus acidophilus for using food industry discarded object as feedstuff with auxiliary material mixing after fermentation, be prepared into the method for the micro-ecological functions feed of ruminant.
Background technology
Micro-ecological functions feed is, by multiple beneficial microorganism, relevant feedstuff is carried out to the feed after fermentation process, and microorganism wherein and metabolite match and nutrition complement to the naturally micro-ecosystem in relevant animal body.The micro-ecological functions feed that utilizes microbial fermentation technology to produce, for developing feed resource, improves efficiency of feed utilization, regulates the interior micro-ecology of animal body average, and substitute antibiotics, prevents and treats some disease, has positive effect.
Raw material for the production of fermented feed is very extensive, from food processing accessory substance up till now of the hydrocarbons such as early stage oil, methyl alcohol, ethanol, discarded object, agricultural crop straw etc.Wherein more to the research report that utilizes grouts to produce fermentation protein feedstuff as raw material, the object of such fermentation is ANFs and the toxin of removing wherein, 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 not with zymophyte by manufacture feeds that ferment such as maize pulp-water, corn whitewashing fiber, sauce grain, manioc wastes.
The problem that present maize pulp-water, corn whitewashing fiber, sauce grain, manioc waste exist aspect feed applications below:
Maize pulp-water refers to the suspension of separating in production of corn starch process, and wherein BOD value, up to 1.5mg/g, is rich in organic substance, and general factory all drains into river, river, lake, has caused serious pollution.According to surveying and determination, the moisture of maize pulp-water reaches 90.01%(and is permitted sword autumn, 2002), the concentrate that maize pulp-water is condensed into moisture 60% left and right 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 10% left and right, crude protein 22～24%, crude fibre 11～13% left and right.Produce the bibliographical information that there is not yet of fermented feed for utilizing whitewashing zein fiber.
The poor slag being left after sauce grain refers to and makes 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 after sauce grain is dried and makes feed conversion, but dry run energy consumption is high.
Manioc waste is to produce the waste material after starch or alcohol with cassava, and the moisture of fresh manioc waste is more than 85.0%, the moisture 10.0～12.0% of the manioc waste after oven dry, protein content 4.0% left and right, crude fiber content 12%.0, palatability is poor, directly lower as the value of feed.
To utilizing food industry discarded object to produce fermented feed, there are reports both at home and abroad.Perhaps 2002 sword autumns have been reported and have utilized geotrichum candidum fermented maize yellow seriflux to produce feed protein powder, but need to dry after press filtration, equipment investment is large again, this technology is still in laboratory research stage (Xu Jianqiu, corn starch wastewater is produced 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, the 4th phase in 2000), zymotic fluid is carried out the separation of bacterium liquid by this Technology Need, 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 be to provide the preferred lactobacillus acidophilus of an a kind of strain for food industry discarded object and auxiliary material mixing after fermentation as micro-ecological functions feed, and provide the method for this micro-ecological functions feed of preparation.
[0012] the LAg12-7 bacterial strain that the present invention refers to, or LAg12-7 lactobacillus acidophilus is: LAg12-7
lactobacillus acidophiluslAg12-7.LAg12-7 bacterial strain provided by the present invention separates gastrointestinal tract mucous from yak, through be accredited as lactobacillus acidophilus (
lactobacillus acidophilus).LAg12-7
lactobacillus acidophiluslAg12-7 lactobacillus acidophilus preservation date: 2012 3 days on the 8th, depositary institution: Chinese Typical Representative culture collection center, depositary institution address: China, Wuhan, Wuhan University (phone 027-68754052), survival detects August 7 2012 date: preserving number is: CCTCC NO:M 2012305; This lactobacillus acidophilus has following characteristic:
(1) this bacterium is inoculated in MAS fluid nutrient medium, cultivates and within 22 hours, reach the highest growth concentration under 37 ℃ 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 36 hours under 37 ℃ of conditions after, the pH value of nutrient solution is shown in Fig. 2 lower than 3.50();
(3) this bacterium under pH2.0 condition, to process the survival rate after 2 hours be that 69.32%(is in table 1);
(4) after the cholate condition that this bacterium is 0.4% concentration in weight ratio is processed 2 hours, 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(is in table 3).
The micro-ecological functions feed of ruminant containing LAg12-7 lactobacillus acidophilus provided by the present invention at least has following a kind of function:
(1) strengthen the feed intake of ruminant to roughage and the digestibility to dietary fiber: the feed intake of vinasse is improved to 4.46%～13.24%(in table 6), crude fiber digestibility improves 9.73%～19.02%(in table 8);
(2) improve ruminant production performance and economic benefit: daily gain improves 7.89%～8.77%(in table 7), gross profit increases by 42.94%～76.69%(in table 11);
(3) there is the function that regulates ruminant tumor gastric microbial balance, increase the quantity of Rumen Cellulolytic Bacteria: the quantity utmost point to yellow Ruminococcus etc., Ruminococcus albus, Lactobacillus acidophilus and bargen's streptococcus significantly improves (in table 9);
(4) protection ruminant body antioxidant system, raising non-specific immune function; strengthen premunition: the concentration to total protein, albumin and the each index of three kinds of immunoglobulin (Ig)s (IgA, IgG and IgM) in serum obviously improves, and the activity of TAC and superoxide dismutase is obviously improved to (in table 10).
The technology contents that the present invention relates to bacterium, microbial inoculum, activation method, feed formula and feed making method is as follows:
1, the acquisition of LAg12-7 lactobacillus acidophilus
Adopt separation, the screening and identification method of conventional lactic acid bacteria, from yak is gastrointestinal tract mucous, separation obtains 108 strains of lactic acid bacteria, measure 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 Chinese Typical Representative culture collection center (Wuhan University's preservation center) preservation, and it is numbered CCTCC M 2012305, 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 Chinese Typical Representative culture collection center (Wuhan University's preservation center) preservation, and it is numbered the LAg12-7 lactobacillus acidophilus of CCTCC M 2012305, and its ice dry powder viable count is 20,000,000,000/gram; Bacillus subtilis freeze-dried powder is purchased from magnificent occasion industrial (Chengdu) Science and Technology Ltd., and its viable count is 20,000,000,000/gram; Saccharomyces cerevisiae freeze-dried powder is purchased from Angel Yeast Co.,Ltd, and 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 to make and close zymocyte liquid.The method that zymocyte liquid is closed in preparation is: take by weight 5.0～15.0% brown sugar and add 80.0～90.0% clean water, 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; Keep temperature and time to be: under 32 ℃ of conditions, be incubated 2～3 days, between soak, stir once every day, and pH can drop to below 4.0, obtain the composite zymocyte liquid having activated.
3, compositing formula and the preparation method of the micro-ecological functions feed of ruminant
Need and the feature of feeding according to ruminant animal nutrition, the fermentation major ingredient that is first 75.0～85.0% by weight ratio mixes with 10.0～20.0% fermentation auxiliary material, add again 2.0～6.0% composite zymocyte liquid to be fully mixed and made into fermentation raw material, then fermentation raw material is packed in fermentation polybag, sealing and fermenting polybag ferments fermentation raw material in fermentation polybag.Fermentation condition is: 15～30 ℃ of condition bottom fermentations of environment temperature 2～3 days, obtain the micro-ecological functions feed of finished product ruminant; Or 6～15 ℃ of condition bottom fermentations of environment temperature 5～7 days, obtain the micro-ecological functions feed of finished product ruminant.
Take the composition of top fermentation major ingredient and weight ratio as: take by weight 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% and directly mix.
Composition and the weight ratio of fermentation auxiliary material are: take by weight wheat bran 50.0～65.0%, and 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: take by weight 5.0～15.0% brown sugar and add 80.0～90.0% clean water, 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; Keep temperature and time to be: under 32 ℃ of conditions, be incubated 2～3 days, between soak, stir once every day, and pH can drop to below 4.0, obtain the composite zymocyte liquid having 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: at present domestic to food processing discarded object maize slurries, whitewashing zein fiber, the method of sauce grain and manioc waste exploitation feed product is single, equipment complexity, 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, add fermentation auxiliary material according to the nutritional need feature of ruminant again, add again the composite zymocyte liquid after activation (to contain LAg12-7 lactobacillus acidophilus, bacillus subtilis and saccharomyces cerevisiae) after, directly pack in fermentation polybag and carry out solid state fermentation.The present invention has the following advantages:
1, good LAg12-7 lactobacillus acidophilus strain
In 108 strains of lactic acid bacteria of the present invention's separation from yak is gastrointestinal tract mucous, obtain the lactobacillus acidophilus of a strain function admirable through repeated screening, called after LAg12-7 lactobacillus acidophilus (microbial preservation numbering: CCTCC M 2012305).This bacterium is inoculated in MRS fluid nutrient medium, within 22 hours, reaches the highest growth concentration 37 ℃ of cultivations, and OD600 value is 2.86, in MRS fluid nutrient medium, under 37 ℃ of conditions, cultivates 36 hours, and the pH value of nutrient solution is lower than 3.50; Survival rate after pH2.0 processes 2 hours is 69.32%; Survival rate after 0.7%% gallbladder salinity is processed 2 hours is 53.60%; Escherichia coli, staphylococcus aureus, salmonella, Shigella all being had comparatively to significantly inhibitory action, 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.
Due to the product acid of LAg12-7 lactobacillus acidophilus, acidproof and bacteriostasis property good, and can there is good compatibility with bacillus subtilis and saccharomyces cerevisiae, pH value in fermented feed was down to rapidly below 4.0 in 24 hours, suppress fast harmful microbe growth, and make the beneficial bacterium state of having the advantage all the time, can make fermented feed energy long term guarantee quality.
2, the micro-ecological functions feed of preparation ruminant is simple for process
This fermented feed changes general solid state fermentation pattern, pass through strain combination, the fermentation raw material preparing is put into fermentation polybag, carry out the aerobic three-dimensional energy-conservation fermentation combining with anaerobism: mainly fermented by aerobic bacteria (saccharomycete) early stage, consume the oxygen in fermentation raw material, later stage is mainly fermented by anaerobic bacteria (lactobacillus acidophilus), pH value in fermented feed is reduced to rapidly below 4.0 with interior for 24 hours, suppress fast harmful microbe growth, bacillus subtilis breeding simultaneously produces the digestive ferments such as a large amount of cellulases, protease.The micro-ecological functions feed of ruminant that gets product fermented through 7～20 days.Polybag that can loading mass 40kg fermentation raw material is round, is also finished product packing, storage bag and shipping bags, need not pack again, need not be dried again, is directly transported to user.
3, the micro-ecological functions Feed Manufacturing of ruminant cost is low
The present invention, take food industry discarded object at a low price as primary raw material, adopts to gurry raw material the energy-conservation zymotechnique that need not dry simultaneously, makes the production cost of feed low.Due to by add suitable fermentation auxiliary material and and ferment containing the composite fermentation microbial inoculum of LAg12-7 lactobacillus acidophilus after, become and there is the micro-ecological functions feed of ruminant that better nutritivity is worth.In ruminant fine fodder, add 20～60% the micro-ecological functions feed of the present invention, in improving ruminant production performance, can reduce ruminant feed cost 4% left and right.
4, the micro-ecological functions feed of ruminant is easy to use
The present invention is according to the nutritional need of ruminant and handling characteristics design, make the nutritive index of the micro-ecological functions feed after fermentation substantially reach the nutritive index of ruminant fine fodder, user can directly substitute ruminant fine fodder with 20～60% weight ratio in use, need not change or adjust original ruminant Concentrate formula, easy to use.
5, the micro-ecological functions feed of ruminant 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, the micro-ecological functions feed of ruminant long shelf-life
The micro-ecological functions feed of ruminant of the present invention is guaranteed the quality 70～90 days under 30 ℃～40 ℃ conditions of environment temperature; Or guarantee the quality 150～180 days under 0～30 ℃ of condition of environment temperature.And the existing feed fermenting containing lactobacillus acidophilus is guaranteed the quality 15～20 days under 30 ℃～40 ℃ conditions of environment temperature; Or guarantee the quality 30～40 days under 0～30 ℃ of condition of environment temperature.
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, screening yak stomach and intestine lactic acid bacteria obtains 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, be settled to 1000ml with distilled water, 6.2 ~ 6.4,115 ℃ of sterilizing 30min of pH.
Adopt conventional micro-biological process, from yak is gastrointestinal tract mucous, separation obtains 108 strains of lactic acid bacteria, by producing acid, acidproof salt tolerant experiment etc., therefrom filters out the bacterial strain of a strain function admirable.This bacterial strain identifies through Physiology and biochemistry and 16SrDNA Molecular Identification determines that this bacterial strain is lactobacillus acidophilus, called after lactobacillus acidophilus LAg12-7, and bacterial strain is made into freeze-dried powder through freeze drying, and the viable count of freeze-dried powder is 20,000,000,000/gram.This bacterium is by Chinese Typical Representative culture collection center (Wuhan University's preservation center) preservation, and it is numbered CCTCC M 2012305.
1.2, LAg12-7 lactobacillus acidophilus growth test
Lactobacillus acidophilus LAg12-7 is proceeded in MRS fluid nutrient medium to 37 ℃ of activation 15～18 little.The bacterial strain having activated is proceeded to anaerobism in MRS fluid nutrient medium with the inoculum concentration of 2% (V/V) and cultivates,, 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, in 2.86 left and right, in growing stationary phase, is shown in Fig. 1.
1.3, LAg12-7 lactobacillus acidophilus produces acid test
By after LAg12-7 lactobacillus acidophilus activation, be inoculated into MRS fluid nutrient medium, under 37 ℃ of conditions, to cultivate, sampling and measuring pH value at regular intervals, take the pH of nonvaccinated MRS fluid nutrient medium as initial pH.The results are shown in Figure 2, after cultivating as seen 36 hours, the pH value of LAg12-7 lactobacillus acidophilus nutrient solution is lower than 3.50.
After LAg12-7 lactobacillus acidophilus activation, inoculation MRS fluid nutrient medium is cultivated under 37 ℃ of conditions, measures zymotic fluid pH after cultivating certain hour, take the pH of nonvaccinated MRS fluid nutrient medium as initial pH.Result shows, this bacterial classification cultivate after pH fast-descending, cultivate 36h after pH downward trend ease up, while cultivating to 60h, pH reaches 3.45. cultivation is after 36 hours as seen, 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
Bacterial classification is seeded on MRS fluid nutrient medium and is activated 2 times, with 2%(V/V) inoculum concentration be inoculated in MRS fluid nutrient medium, under 37 ℃ of conditions, cultivate 24h hour, centrifugal 10 minutes of 8000rpm/min, collect thalline, by thalline pbs(pH6.0) buffer solution washing 1 time, Eddy diffusion is in pbs(pH6.0) in buffer solution.Suspension is divided into 5 points, again centrifugal collection thalline.The thalline obtaining is suspended in respectively in pH2.0,3.0,4.0,5.0 and 6.0 pbs buffer solution, the pbs bacterium liquid of 5 parts of different pH that obtain, at room temperature places 2 hours, carries out count plate, take the viable count under pH6.0 condition as 100%, calculate its survival rate.
LAg12-7 lactobacillus acidophilus acid resistance test the results are shown in Table 1, shows that the survival rate that LAg12-7 lactobacillus acidophilus was placed after 2 hours under pH2.0 condition is 69.32%.
The survival rate (unit: %) of table 1 LAg12-7 lactobacillus acidophilus under different pH values
1.5, LAg12-7 lactobacillus acidophilus bile tolerance test
Cholate addition is established 4 gradients: 0.1%, 0.3%, 0.5% and 0.7%, and take the culture medium that do not add cholate as contrast.Bacterial classification is seeded on MRS fluid nutrient medium and is activated 2 times, be inoculated in treatment samples and control sample with 2% inoculum concentration, under 37 ℃ of conditions, leave standstill to cultivate after 2 hours and carry out count plate.
LAg12-7 lactobacillus acidophilus bile tolerance result of the test, in table 2, shows that the survival rate that LAg12-7 lactobacillus acidophilus was cultivated after 2 hours under 0.7% cholate condition is 53.60%.
The survival rate (unit: %) of table 2 lactic acid bacteria under different salinity
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 15mL LB solid medium, after culture medium solidifying, to place diameter be 7mm Oxford cup, adds the fermented supernatant fluid of lactobacillus acidophilus LAg12-7, at room temperature spreads 30min, under 37 ℃ of conditions, cultivate 16-18 hour, observe inhibition zone.Result is as shown in table 3.
The antibacterial circle diameter (unit: mm) of table 3 LAg12-7 lactobacillus acidophilus
The method of the composite fermentation microbial inoculum that embodiment 2, the micro-ecological functions feed of preparation ruminant are used
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 and just make the composite fermentation microbial inoculum that the micro-ecological functions feed of ruminant is used under normal temperature condition.
The method of the composite fermentation microbial inoculum that embodiment 3, the micro-ecological functions feed of preparation ruminant are used
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 and just make the composite fermentation microbial inoculum that the micro-ecological functions feed of ruminant is used under normal temperature condition.
The method of the composite fermentation microbial inoculum that embodiment 4, the micro-ecological functions feed of preparation ruminant are used
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 and just make the composite fermentation microbial inoculum that the micro-ecological functions feed of ruminant is used under normal temperature condition.
The method of the composite fermentation microbial inoculum that embodiment 5, the micro-ecological functions feed of preparation ruminant are used
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 and just make the composite fermentation microbial inoculum that the micro-ecological functions feed of ruminant is used under normal temperature condition.
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 the activation of composite fermentation microbial inoculum: 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; Keep temperature and time to be: under 32 ℃ of conditions, be incubated 2～3 days, between soak, stir once every day, and pH can drop to below 4.0, obtain the composite zymocyte liquid having 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 of composite fermentation microbial inoculum activation is same as to 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 of composite fermentation microbial inoculum activation is same as to 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 of composite fermentation microbial inoculum activation is same as to embodiment 6.
The preparation method of embodiment 10, the micro-ecological functions feed of ruminant
In the micro-ecological functions feed of ruminant, 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%;
Composition and the weight ratio of fermentation auxiliary material 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%;
Raw material composition and the weight ratio of the micro-ecological functions feed of ruminant 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 of making the micro-ecological functions feed of ruminant of above-mentioned raw materials is as follows:
(1) make fermentation major ingredient: take by weight maize slurries, whitewashing zein fiber, sauce grain and manioc waste and directly mix;
(2) make fermentation auxiliary material: take by weight wheat bran, beer yeast powder, calcium carbonate, sodium acid carbonate, magnesium chloride directly mix;
(3) make composite zymocyte liquid: take by weight brown sugar 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; Keep temperature and time to be: under 2 ℃ of conditions, be incubated 2～3 days, between soak, stir once every day, and pH can drop to below 4.0, obtain the composite zymocyte liquid having activated;
(4) make functional feed: be first that fermentation major ingredient mixes with fermentation auxiliary material by weight ratio, add again composite zymocyte liquid to be fully mixed and made into fermentation raw material, then fermentation raw material is packed in fermentation polybag, sealing and fermenting polybag ferments fermentation raw material in fermentation polybag; Fermentation condition is: 15～30 ℃ of condition bottom fermentations of environment temperature 2～3 days, obtain the micro-ecological functions feed of finished product ruminant; Or 6～15 ℃ of condition bottom fermentations of environment temperature 5～7 days, obtain the micro-ecological functions feed of finished product ruminant.
The preparation method of embodiment 11, the micro-ecological functions feed of ruminant
In the micro-ecological functions feed of ruminant, 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%;
Raw material composition and the weight ratio of the micro-ecological functions feed of ruminant 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 of making the micro-ecological functions feed of ruminant of above-mentioned raw materials is same as embodiment 10.
The preparation method of embodiment 12, the micro-ecological functions feed of ruminant
In the micro-ecological functions feed of ruminant, 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 of composite fermentation microbial inoculum activation is same as to embodiment 6;
Raw material composition and the weight ratio of the micro-ecological functions feed of ruminant 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 of making the micro-ecological functions feed of ruminant of above-mentioned raw materials is same as embodiment 10.
The preparation method of embodiment 13, the micro-ecological functions feed of ruminant
In the micro-ecological functions feed of ruminant, 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 of composite fermentation microbial inoculum activation is same as to embodiment 6;
Raw material composition and the weight ratio of the micro-ecological functions feed of ruminant 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 of making the micro-ecological functions feed of ruminant of above-mentioned raw materials is same as embodiment 10.
Embodiment 14, the micro-ecological functions feed of beef cattle feeding experiment
The present embodiment adds 25% or 40% the micro-ecological functions feed of ruminant of the present invention in fattening beef cattle fine fodder, can increase the feed intake 4.46%～13.24% of beef cattle to coarse fodder, 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, thereby increase economic efficiency (gross profit) 42.94%～76.69%.From this sample result, take the effect of adding micro-ecological functions feed of 40% in basic fine fodder as good.The mensuration of the indexs such as concrete feeding experiment and the production performance to test beef cattle, nutrient digestion, rumen microorganism, Physiology and biochemistry and immunity below:
1, testing program
1.1, the micro-ecological functions feed of beef cattle: with the micro-ecological functions feed of ruminant containing lactobacillus acidophilus of the present invention.
1.2, experimental design
Test is carried out in certain large-scale beef cattle plant.Select 90 of the consistent assorted oxen in west of body weight 373.40 ± 42.53kg, body condition hair color, adopt 5 × 3 RANDOMIZED BLOCK DESIGN, according to body weight phase approximately principle, 90 oxen are divided into 5 district's groups, 18 of each district groups, 18 oxen of same district group are divided into individual processing (control group, test I group and II group), 5 repetitions of each processing, 6 oxen of each repetition randomly; Control group fed basis fine fodder, test I group and II group substitute basic fine fodder with micro-ecological functions feed with 25% and 40% ratio respectively, respectively organize fine fodder (4kg) and the corn stalk pellets (2.7kg) of the identical weight of feeding every every day, vinasse free choice feeding, fasten and feed, freely drink water; Test period: raise in advance 10 days phases, just trying 48 days phases; Record feed intake every day, every ox in morning of on-test and end is weighed on an empty stomach, eachly when off-test repeat to select 3 oxen that approach this repetition average weight to butcher; to get longissimus dorsi muscle and measure meat index; get rumen fluid and measure the quantity of rumen microorganism, serum biochemistry and immune indexes are measured in jugular vein blood sampling.
1.3, feed formula and nutritional labeling
Control group (basic fine fodder) is original fine fodder in field, and test I group substitutes 25% of basic fine fodder weight with the micro-ecological functions feed of ruminant containing lactobacillus acidophilus of the present invention, tests 40% of basic fine fodder weight of II group generation.The weight ratio formula of each processed group fine fodder is in table 4.
The composition of the 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 |
| Micro-ecological functions 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
The feed every day corn stalk pellets feed of 4kg fine fodder and 1.7kg of every beef cattle, alcohol free choice feeding, records the vinasse feed intake of each repetition every day, calculates every average feed intake of full phase of beef cattle; When on-test and end, every beef cattle is weighed morning on an empty stomach, calculates the average daily gain of every beef cattle.
1.4.2, nutrient digestibility
First three day of off-test gathers Feed Sample and excrement sample, adopts 4N HCl 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 is got rumen fluid after butchering, and adopts RT-PCR absolute quantitation method to measure the quantity of rumen microorganism, and four kinds of microorganisms measuring and 16Sr DNA fragmentation primer sequence thereof are as table 5.
The increase 16Sr DNA fragmentation primer sequence of yellow Ruminococcus, Ruminococcus albus, lactic acid bacteria and bargen's streptococcus of table 5
1.4.3, the mensuration of Biochemical Indices In Serum, immune indexes and Antioxidant Indexes
Jugular vein blood 20ml, leaves standstill after half an hour 3000prm centrifugal 15 minutes under room temperature, make a serum-20 ℃ refrigerator and recommend with guarantee stand-by.Total serum protein (BCA method), albumin (bromocresol green method), glutamic-pyruvic transaminase/glutamic-oxalacetic transaminease (reitman-frankel method), immunoglobulin (Ig) (IgA, IgG and IgM), TAC, total superoxide dismutase 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, the impact of the micro-ecological functions feed of ruminant on beef cattle production performance
2.1.1, the impact of the micro-ecological functions feed of ruminant on beef cattle feed intake
Every of each processing ox fine fodder 4 kg/ heads of feeding every day. day, corn stalk pellets 2.7 kg/ heads, vinasse free choice feeding.Each group feed intake the results are shown in Table 6.As shown in Table 6, test I group and II group than control group search for food vinasse 0.89kg and 0.30kg, improve respectively 13.24% and 4.46% respectively more, but not remarkable (P > 0.05) of difference.
The impact of the micro-ecological functions feed of table 6 on beef cattle feed intake
2.1.2, daily gain
The daily gain of each processed group is in table 7.Result shows, duration of test is tested I group and every beef cattle of II group respectively than many control groups many weightening finishes 4.59kg and 4.91kg, and daily gain you can well imagine high 7.89% and 8.77% than contrast component, but respectively organizes not remarkable (P > 0.05) of difference.
The impact of the micro-ecological functions feed of table 7 on beef cattle daily gain
2.2, the micro-ecological functions feed of ruminant is on the impact on cattle diet nutrient digestibility
The nutrient digestibility of each processed group changes in table 8.Result shows, adds micro-ecological functions feed in fine fodder, has the trend that increases crude protein digestibility, but not remarkable (P > 0.05) of difference; The utmost point significantly improves coarse-fibred digestibility (P < 0.01).
The impact (unit: %) of the micro-ecological functions feed of table 8 on cattle diet nutrient digestibility
2.3, the impact on 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, micro-ecological functions feed increases the quantity of Rumen Cellulolytic Bacteria (yellow Ruminococcus and Ruminococcus albus) and Lactic Acid Producing (Lactobacillus acidophilus and bargen's streptococcus), and has the trend of increase with the increase of micro-ecological functions feed addition.
Impact (the unit: × 10 of the micro-ecological functions feed of table 9 on rumen microorganism quantity
8copy number/μ l, %)
2.4, the impact on the growth of beef cattle serum, immunity and Antioxidant Indexes
Micro-ecological functions feed on the impact of the growth of beef cattle serum, 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 significantly lower than control group; But all not significantly (P > 0.05) of the Biochemical Indices In Serum difference between two test group.
Immunoglobulin IgA, IgG and IgM in serum, Zong and TAC and the superoxide dismutase content utmost point be significantly higher than control group (P < 0.01), and be increase trend with the increase of micro-ecological functions feed addition; And test group mda content is extremely significantly lower than control group (P < 0.01).
The impact of the micro-ecological functions feed of table 10 on 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 that forms 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 be removed ultra-oxygen anion free radical, and Cell protection is avoided damage, and the oxidative and anti-oxidative balance of body is played to vital effect.MDA is the end-product of the MDA that produces of peroxidatic reaction of lipid that oxygen radical causes at body intracellular metabolite, can indirectly reflect degree and level of lipid peroxidation that situation, the body tissue cell of radical metabolism in body are attacked by free radical.
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 superoxide dismutase.Micro-ecological functions Feed Energy protection beef cattle body antioxidant system is described, improves 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 with 0. 80 yuan, according to the feed intake of table 6, the feed cost that calculates every every day of each processed group 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 according to the daily gain of table 7, the income that calculates every every day of each processed group is respectively 18.81 yuan, 20.30 yuan and 20.46 yuan.Than 1.63 yuan/day of control groups. 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 micro-ecological functions feed of the ruminant 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 micro-ecological functions 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 Chinese Typical Representative culture collection center (Wuhan University's preservation center) preservation, and its deposit number is CCTCC M 2012305.
2. the preparation method of the micro-ecological functions feed of ruminant according to claim 2, is characterized in that this feed making method is as follows:
(1) make fermentation major ingredient: take by weight 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% and directly mix;
(2) make fermentation auxiliary material: take by weight 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% directly mix;
(3) make composite zymocyte liquid: take by weight 5.0～15.0% brown sugar and add 80.0～90.0% clean water, 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; Keep temperature and time to be: under 32 ℃ of conditions, be incubated 2～3 days, between soak, stir once every day, and pH can drop to below 4.0, obtain the composite zymocyte liquid having activated;
(4) make functional feed: the fermentation major ingredient that is first 75.0～85.0% by weight ratio mixes with 10.0～20.0% fermentation auxiliary material, add again 2.0～6.0% composite zymocyte liquid to be fully mixed and made into fermentation raw material, then fermentation raw material is packed in fermentation polybag, sealing and fermenting polybag ferments fermentation raw material in fermentation polybag; Fermentation condition is: 15～30 ℃ of condition bottom fermentations of environment temperature 2～3 days, obtain the micro-ecological functions feed of finished product ruminant; Or 6～15 ℃ of condition bottom fermentations of environment temperature 5～7 days, obtain the micro-ecological functions feed of finished product ruminant.
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| CN111202177A (en) * | 2020-01-17 | 2020-05-29 | 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 |
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103798503B (en) | 2015-11-18 |
| CN102943053A (en) | 2013-02-27 |
| CN103881944B (en) | 2016-03-30 |
| CN103881944A (en) | 2014-06-25 |
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