CN105199978A - Bacillus coagulans preparation for livestock breeding and preparation method thereof - Google Patents
Bacillus coagulans preparation for livestock breeding and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a bacillus coagulans preparation for livestock breeding and a preparation method thereof and belongs to the technical field of additives for livestock breeding. The preparation is prepared through the following steps: 2-5% of soluble starch, 0.5-1% of yeast extract, 0.5-1.5% of peptone, 1.0-1.5% of soybean meal and the balance of water are adopted to prepare a fermentation medium; with the BFS-2 strains of the bacillus coagulans with the preservation number of CCTCC No.M 2011422 as strains, carrying out all-stage fermentation, so as to obtain the bacillus coagulans preparation. The preparation is low in price, and high in content of bacterial count, the guarantee period is one year or above, and after the preparation is used, the growth of harmful bacteria in the intestines of animals can be effectively restrained, the micro-ecological environment of the intestines tracts of bred animals is improved, and the incidence of diarrhea is reduced; meanwhile, the utilization of the feed is effectively promoted, the breeding cost is reduced, the culture benefits are increased and the preparation can be popularized and applied in livestock farms.
Description
Technical field
The present invention relates to livestock and poultry cultivation additive agent field, particularly relate to a kind of microbial preparation-Bacillus coagulans for livestock and poultry cultivation and preparation method thereof.
Background technology
In recent years, along with aquaculture is intensive, the fast development of mass-producing, the outburst of cultivated animals disease is more and more frequent, endangers also increasing, and cultivated animals disease has become the bottleneck of restriction aquaculture development.
The application of the chemical agents such as microbiotic in aquaculture has the history of decades, and to prevention Animal diseases, the seed output and quality improving aquaculture serves positive effect.But along with deepening continuously of understanding, it is found that life-time service microbiotic can destroy the micro-flora in livestock and poultry alimentary tract, animal body is developed immunity to drugs to microbiotic, and microbiotic remaining in animal products, directly have influence on the health of the mankind and the safety of ecotope.Therefore find the antibiotic substitute that can overcome above-mentioned drawback, the green feed additive of development of new safety, become the focus that Chinese scholars is paid close attention to and studied.
Micro-ecological additive is a kind of a kind of Multifunctional Compound Microorganism Product adopting nutrition slow-release technology, biofermentation technique and special producing technique to cultivate, it both can not develop immunity to drugs, the problem such as drug residue free again, be considered to the fodder additives replacing microbiotic most potentiality, obtain increasing attention.Current milk-acid bacteria, genus bacillus are the probioticses be most widely used.Milk-acid bacteria can provide available indispensable amino acid and various VITAMIN for host, has the effect of direct growth promoting effects.Genus bacillus can produce proteolytic enzyme, lipase, Zulkovsky starch enzyme isoreactivity material, antibiotic can also be secreted, there is comparatively Johnson & Johnson's thing in animal intestinal and take oxygen ability by force, to the growth promoting the useful anaerobion such as milk-acid bacteria, bifidus bacillus in animal body, maintain microecological balance and play an important role.Therefore, the research of the bacillus JY-LZ of both advantages is concentrated to become the focus of probiotic strain research and development in recent years.The Bacillus coagulans having obtained more application in developed country is most important one in feeding bacillus JY-LZ, it not only has all features of milk-acid bacteria, owing to producing gemma, it also has all uniquenesses of genus bacillus, is one of current good animal health kind.As far back as 1989, U.S. FDA just disclosed the safe bacterial strain that 41 kinds can be used for feed, and within 1992, again ratify 42 kinds, wherein Bacillus coagulans makes number one.The research and development of China to Bacillus coagulans are started late, and the research of recent year to Bacillus coagulans focuses mostly in medical science, and has achieved good curative effect.Be used for the treatment of enteritis, diarrhoea, constipation, maldigestion etc. that enteric flora disturbance causes, Be very effective.Therefore, develop function strong, cost livestock and poultry cultivation probiotics low, with strong points has become the emphasis of research both at home and abroad.
Summary of the invention
The object of the invention is, utilize the high-efficiency strain screened from physical environment, use statistical method and modern biotechnology fermentation engineering and means, carry out the aftertreatment complete processing of low cost, high-density industrialization production and advanced person, aquaculture is applied to as antibiotic surrogate, to promotion livestock industry volume increase, synergy and healthy and sustainable development, there is significant economic benefit and DEVELOPMENT PROSPECT.Another object of the present invention is to provide a kind of preparation method of this microbial preparation.
The object of the invention is achieved through the following technical solutions.
For a Bacillus coagulans preparation for livestock and poultry cultivation, wherein, said preparation comprises and is deposited at Wuhan, China, and China typical culture collection center preservation, deposit number is the BFS-2 bacterial strain of CCTCCNo:M2011422.
Plant Bacillus coagulans (Bacilluscoagulans) BFS-2 bacterial strain:
We are separated the microorganism strains had compared with high acid ability from the different sourcess such as weanling pig enteron aisle, vegetable soil, pickles, and in conjunction with morphological specificity, the physiological and biochemical index of bacterium, the modes such as 16SrRNA Molecular Identification are identified the high efficient strain obtained; And on this basis, initial analysis is carried out to the bacteriostatic activity of the Bacillus coagulans obtained.Result shows, is altogether separated and obtains acid-producing bacteria strain 43 strain from above-mentioned various different sources, after producing acid test and acidproof, bile tolerance Preliminary Experiment and screening, determines 3 strain microorganism strains.Biochemical and the Molecular Identification through subsequent physiological, this 3 strain is Bacillus coagulans.Antibacterial result shows, the inhibition of this 3 bacillus coagulans to gemma genera bacillus is poor, better to the suppression situation of the enteron aisle conditioned pathogens such as intestinal bacteria, the bacterial strain that wherein net effect is best is BFS-2, and the sampling of this bacterial strain is separated from a vegetable garden of Zhejiang Hangzhou.
The feature such as form, cultivation, physiology, biochemistry of preservation strain:
Bacillus coagulans BFS-2 bacterial strain: belong to genus bacillus (Bacilluscoagulans); Gram-positive microorganism, well-grown in MRS liquid nutrient medium, cell is shaft-like, has gemma; When MRS agar plate is cultivated, bacterium colony is circular, neat in edge, rule, canescence, and surface is glossy.
This BFS-2 bacterial strain on November 25th, 2011, at Wuhan, China, China typical culture collection center preservation, deposit number: CCTCCNo:M2011422.
On the other hand, the invention provides a kind of microbial preparation for the preparation of livestock and poultry cultivation-Bacillus coagulans preparation,
Said preparation can obtain by the following method: with potato glucose solid medium for slant medium; Take potato dextrose broth as shaking flask bacterium culture medium; Zulkovsky starch 2-5% by weight, yeast extract paste 0.5-1%, peptone 0.5-1.5%, dregs of beans 1.0-1.5%, water complements to 100% and is mixed with seeding tank and ferment tank substratum; Take deposit number as Bacillus coagulans (Bacilluscoagulans) the BFS-2 bacterial strain of CCTCCNo:M2011422 be bacterial classification, after fermentation at different levels, be prepared from viable count is 1X10
8the Bacillus coagulans of CFU/g.
For the preparation method of livestock and poultry cultivation microbial preparation-Bacillus coagulans preparation, carry out according to the following steps:
(1) preparation of substratum at different levels, wherein:
A) slant medium: potato glucose solid medium, for subsequent use;
B) shaking flask bacterium culture medium: potato dextrose broth, for subsequent use;
C) seeding tank fermention medium: Zulkovsky starch 2-5% by weight, yeast extract paste 0.5-1%, peptone 0.5-1.5%, dregs of beans 1.0-1.5%, water complements to 100%;
D) fermentation tank culture medium: Zulkovsky starch 2-5% by weight, yeast extract paste 0.5-1%, peptone 0.5-1.5%, dregs of beans 1.0-1.5%, water complements to 100%;
(2) cultivation of slant strains: by preservation of bacteria strain: CGMCCNo.2288 Bacillus coagulans BFS-2 inoculation is on step (1) slant medium, cultivate 18-24 hours in the incubator of 28-37 DEG C, directly apply or put into refrigerator and cooled after taking-up to hide, for subsequent use;
(3) shaking flask bacterial classification amplification: under aseptic technique, gets in ring slant strains access 60ml step (1) shaking flask bacterium culture medium, in 28-37 DEG C, in the shaking table of 110-220 revs/min, cultivate 18-24 hours, take out and directly apply or put into refrigerator and cooled Tibetan, for subsequent use;
(4) seeding tank fermentation: step (1) seeding tank fermention medium is put into seeding tank, in seeding tank, temperature 121 DEG C kept after 30 minutes, and pressurize, to 0.1MPa, is cooled to 28-37 DEG C after carrying out steam high-temperature sterilization; Under flame protection, access step (3) shaking flask bacterial classification rapidly by inoculation valve in seeding tank, inoculum size is 5-10%; Start stirrer rotating speed 110-220 revs/min, temperature 28-37 DEG C, air flow 4m
3/ h, tank pressure 0.02-0.06MPa, pH nature, cultivates 18-24 hours, for subsequent use;
(5) preparation of bulk article: step (1) ferment tank substratum is put into fermentor tank, temperature 121 DEG C in fermentor tank, pressurize, to 0.1MPa, keeps, after 30 minutes, substratum is cooled to 28-37 DEG C; In fermentor tank, access step (4) seeding tank bacterial classification by culture transferring pipeline, inoculum size is 5-10%; Start stirrer rotating speed 110-220 revs/min, temperature 28-37 DEG C, air flow 20m
3/ h, tank pressure 0.02-0.06MPa, pH nature, cultivates 18-24 hours, inspection by sampling, aftertreatment (product form needs to be produced into solid or liquid according to client) is carried out according to target level of product quality, filling, sealing, for the microbial preparation of livestock and poultry cultivation, wherein, the gemma viable count that condenses is 1X10
8cFU/g.
The application method of livestock and poultry cultivation microbial preparation-Bacillus coagulans: the consumption interpolation microbial preparation-Bacillus coagulans in feed per ton being 0.005-0.1% by mass percentage, Direct-fed after stirring, wherein, said preparation comprises and is deposited at Wuhan, China, China typical culture collection center preservation, deposit number is the BFS-2 bacterial strain of CCTCCNo:M2011422 or the cultured products of this bacterial strain.Here concentration can be 0.005%, 0.05%, any ratio in 0.1%.
In some preferred modes, the mass percent of Bacillus coagulans is 0.05%.
In some preferred modes, condensation gemma viable count is 1X10
8-5X10
8cFU/g.
For the biological characteristics explanation of preservation bacterium
This bacterial isolates BFS-2 is preserved in China typical culture collection center preservation, preservation number is CCTCCNo:M2011422, address: Wuhan, China, Classification system: Bacilluscoagulans, Chinese is: Bacillus coagulans, preservation date: on November 25th, 2011.
Beneficial effect of the present invention
1, present invention employs the fermentation pattern of liquid fermenting, with the Bacillus coagulans BFS-2 bacterial strain of preservation for cultivating bacterial classification, and by testing the behavioral study of this bacterium and the impact of each factor, determine C, N ratio be applicable to, the formula of growth temperature, inoculum size and substratum, is aided with optimum preparation technology and method, and institute's bacteriostatic agent product effectively can suppress the growth of harmful bacteria in cultivated animals enteron aisle, the intestine microenvironment of improvement cultivated animals, reduces Incidence of Diarrhea; Effectively can promote the utilization of feed simultaneously, reduce aquaculture cost, improve culture benefit;
2, in 21 age in days Du × long × large weanling pig, add the Bacillus coagulans that concentration is 0.05% and 0.1% respectively, do not add microbiotic, carry out the feeding experiment of 52d by a definite date.Test-results shows: compared with control group, and add condensation bud pole bacterium in daily ration after, the day weight gain of weanling pig, feed conversion rate are significantly higher than fed control group piglet, and feedstuff-meat ratio raising reaches 4.97% and 4.48% respectively; Diarrhea rate significantly reduces, and comparatively control group reduces 54.95% and 44.55% respectively.
70 age in days Du × length × Dasanyuan hybridization growing swine are tested, and result shows: add Bacillus coagulans and have no significant effect the growth performance of growing swine and Biochemical Indices In Serum; But the lactobacillus quantity that can effectively improve in ight soil.
3, product of the present invention is raw materials used abundant, cheap, one ton of product price about 5000 yuan, comparatively about the cheap half of imported product; The quality of product is also more stable, more than effective a year and a half quality guaranteed period.
Embodiment
Be described in more detail the present invention by following examples, but following examples are only illustrative, the present invention is not by the restriction of these embodiments.
Explanation to involved material:
Glucose: commercially available;
Peptone: commercially available;
Zulkovsky starch: commercially available;
Yeast extract paste: commercially available;
Dregs of beans: commercially available;
The formula of embodiment 1:(product of the present invention and preparation method 1)
For microbial preparation-Bacillus coagulans of livestock and poultry cultivation, step preparation by the following method:
(1) preparation of substratum at different levels, wherein:
A) PDA inclined-plane solid medium: potato 200g, glucose 2g, agar 2g, 1 liter, water, for subsequent use;
B) shaking flask liquid nutrient medium: 1 liter, potato 200g, glucose 2g, water, for subsequent use;
C) seeding tank fermention medium: Zulkovsky starch 20 grams, yeast extract paste 7 grams, peptone 10 grams, dregs of beans 10 grams, 953 grams, water, for subsequent use;
D) ferment tank substratum: Zulkovsky starch 20 grams, yeast extract paste 7 grams, peptone 10 grams, dregs of beans 10 grams, 953 grams, water, for subsequent use;
(2) cultivation of slant strains: by deposit number: CCTCCNo:M2011422 Bacillus coagulans BFS-2 inoculation is cultivated 24 hours on step (1) PDA inclined-plane solid medium in the incubator of 28 DEG C, take out and directly apply or put into refrigerator and cooled Tibetan, for subsequent use;
(3) preparation of shaking flask liquid spawn: under aseptic technique, get in ring Bacillus strain BFS-2 slant strains access 60ml step (1) shaking flask liquid spawn culture medium, in 28 DEG C, in the shaking table that rotating speed is 220 revs/min, cultivate 24 hours, take out and directly apply or put into refrigerator and cooled Tibetan, for subsequent use;
(4) seeding tank fermentation: preparation steps (1) seeding tank fermention medium also puts into seeding tank, and logical steam makes temperature in seeding tank rise to 121 DEG C, and pressurize is to 0.1MPa, keeps stopping after 30 minutes, subsequently substratum is cooled to 30 DEG C; Accessed in seeding tank by inoculation valve rapidly under flame protection and cultivate qualified step (3) shaking flask liquid seeds bacterial classification through inspection, inoculum size is 6%; Start stirrer rotating speed 180 revs/min, temperature remains on 30 DEG C, air flow 4m
3/ h, tank pressure 0.02MPa, pH nature, incubation time is 24 hours;
(5) preparation of bulk article: preparation steps (1) ferment tank substratum also puts into fermentor tank, logical steam makes temperature in fermentor tank rise to 121 DEG C, and pressurize is to 0.1MPa, keeps stopping after 30 minutes, subsequently substratum is cooled to 30 DEG C; In seeding tank, access cultured step (4) seeding tank liquid seeds bacterial classification by culture transferring pipeline, inoculum size is 6%; Start stirrer rotating speed 180 revs/min, temperature remains on 30 DEG C, air flow 20m
3/ h, tank pressure 0.02MPa, pH nature, cultivated after 24 hours; Be up to the standards and fillingly can make livestock and poultry cultivation microbial preparation-Bacillus coagulans, viable count is 1X10
8cFU/g.The formula of embodiment 2:(product of the present invention and preparation method 2)
The preparation of the present embodiment seeding tank and ferment tank substratum: Zulkovsky starch 30 grams, yeast extract paste 5 grams, peptone 5 grams, dregs of beans 15 grams, 945 grams, water are for subsequent use; All the other preparation methods are all same as embodiment 1.Viable count is 2X10
8cFU/g.
The formula of embodiment 3:(product of the present invention and preparation method 3)
The preparation of the present embodiment seeding tank and ferment tank substratum: Zulkovsky starch 28 grams, yeast extract paste 5 grams, peptone 5 grams, dregs of beans 12 grams, 950 grams, water are for subsequent use.All the other preparation methods are all same as embodiment 1.Viable count is 5X10
8cFU/g.
The effect test of test example 4:(product of the present invention in weanling pig cultivation)
Due to antibiotic a large amount of abuse, its resistance in animal and drug residue problem become increasingly conspicuous.Probiotics, as a kind of novel green fodder additives, with noresidue, pollution-free and have no side effect, and can strengthen animal disease resistant power, promotes growth of animal and improve the advantages such as efficiency of feed utilization and enjoy favor.
Test main researching condensed grafting bud pole bacterium to the impact of Growth Performance of Weaning Piglets, in production, rationally application provides reference.
1 materials and methods
1.1 experimental animals and grouping
White for Du × length of 90 about 8kg × great Bai original seed pig is divided into 3 groups at random, often organizes 3 repetitions, each repetition 10.Daily ration based on the conventional daily ration fed in pig farm, control group fed basal diet, process 1 to be condensed bud pole bacterium+basal diet group for interpolation 0.05%; Process 2 is 0.1% condensation bud pole bacterium+basal diet group.Testing program is in table 1.Condensation bud pole bacterium BFS-2 is by this laboratory screening and fermentation is standby according to the method for examples of implementation 1.Condensation bud pole bacteria preparation form is meal, and main component is condensation bud pole bacterium, and viable count is 1X10
8cFU/g.
The formula of table 1 basal diet and trophic level
1 Preblend provides Cu200mg, Fe180mg, Zn170mg, Mn50mg, Co1.0mg, Se0.4mg for every kilogram of diet, I0.4mg, VA8330IU, VD31440IU, VE30IU, VB2.0mg, VB8.0mg, VB61.2mg, VB0.03mg, folic acid 2.0mg, vitamin H 0.2mg, nicotinic acid 40mg, pantothenic acid 20mg.
2 trophic level are calculated value.
1.2 feeding and management
Undertaken by pig farm Routine Management.Preliminary trial period 5d, carries out preventive vaccination, expelling parasite simultaneously.Enter and just try after date, each component feeds corresponding daily ration.Free choice feeding, freely drinks water, and carries out immunity by pig farm normal procedure.Trial period 52d.
1.3 growth performance testing indexs
Day weight gain: weigh, according to initial weight and last re-computation average daily gain on an empty stomach respectively at the 1d of on-test and last 1d 8:00 in morning.Food consumption and feed-weight ratio: the food consumption calculating weekly each group, per day food consumption, calculates feed-weight ratio after off-test.
Diarrhea rate: piglet ight soil situation is observed in the timing of every morning 8:00 point.Record diarrhoea head number, calculates diarrhea rate.
Diarrhea rate (%)=number of times of always suffering from diarrhoea/(total head number × test number of days) × 100
1.4 serum biochemistry immune indexes measure
The 20:00 testing last 1d stops feeding, as usual supplies water, and carry out precaval vein blood sampling 10mL morning next day to empty stomach piglet, be collected in centrifuge tube, do not add antithrombotics, room temperature leaves standstill 2h, treats that serum is separated out, and sucking-off serum is placed in-20 DEG C of preservations.Adopt automatic clinical chemistry analyzer to measure serum immunoglobulin G (IgG), immunoglobulin A (IgA), immunoglobulin M (IgM) content and complement component 3 (C3), complement 4 (C4) level, mensuration test kit all builds up Bioisystech Co., Ltd purchased from Nanjing.
1.5 enteric microorganism detect
Raising terminates front 3d, 2h after each feeding, gather fresh excreta sample, immediately adopt physiological saline carry out 10 diluted after, get the total milk-acid bacteria of 1mL dilute sample coating MRS flat board detection respectively, coating intestinal bacteria qualification culture medium flat plate detects intestinal bacteria, coating EC bifidus bacillus identifies that substratum detects bifidobacteria.Bacterium number represents with log (CFU/g).
1.6 data statistics
Adopt SPSS statistical software to carry out variance analysis and multiple comparisons to data, multiple comparisons adopts Duncan method, take P<0.05 as conspicuous level.
2 results and discussion
2.1 Bacillus coagulanses are on the impact of Growth Performance of Weaning Piglets
In this test (table 2), adding concentration is after the Bacillus coagulans BFS-2 of 0.05% and 0.1%, and compared with the piglet of fed control daily ration, day weight gain, last weight average significantly improve; Feed weightening finish reduces than remarkable, but not remarkable on the impact of food consumption.This may be because Bacillus coagulans to enter after enteron aisle harmful miscellaneous bacteria in cleaning enteron aisle rapidly, thus promotes that anaerobism probiotics is as the growth of lactobacillus, bifidus bacillus etc., reduces pH value in enteron aisle, suppresses harmful bacteria growth, reaches the effect promoting piglet growth.This is consistent with the influence research result of Bacillus coagulans to weanling pig growth perfonnance of forefathers' bibliographical information.
Table 2 adds the impact of Bacillus coagulans BFS-2 on Growth Performance of Weaning Piglets
2.2 Bacillus coagulanses are on the impact of weanling pig Biochemical Indices In Serum
The impact of Bacillus coagulans on weanling pig Biochemical Indices In Serum the results are shown in Table 3.As seen from table, add in feed concentration be 0.05% Bacillus coagulans can significantly improve the content of IgG in weanling pig serum, but not remarkable on the impact of IgA and IgM.Add concentration be 0.1% Bacillus coagulans all not remarkable on the impact of IgA, IgG and IgM.Meanwhile, add Bacillus coagulans to be significantly increased the level of Complement C_3 and C4.More than research shows that condensation bud pole bacterium can affect the immunity function of animal.Research shows, bud embrace quasi-microorganism can active adsorption at small intestinal mucosal surface, make pathogenic bacteria be difficult to field planting in animal intestinal by biological antagonist and immunization, enteron aisle local and the function of whole body probiotic bacterium immunity can be improved.
Table 3 genus bacillus BFS-2 is on the impact of weanling pig Biochemical Indices In Serum
2.3 Bacillus coagulanses are on the impact of microorganism species in weanling pig ight soil
The impact of Bacillus coagulans on weanling pig Biochemical Indices In Serum the results are shown in Table 4.Have table known, the condensation bud pole bacterium of interpolation 0.05% and 0.1% can increase the quantity (P<0.05) of milk-acid bacteria and bifidus bacillus in ight soil, reduces intestinal bacteria quantity (P<0.05).Test-results and forefathers study basically identical, add 0.05% condensation bud pole bacterium and show good growth promotion and prevent and treat diarrhoea effect in daily ration.Test finds, condensation bud pole bacterium addition is increased to 0.1% from 0.05%, the most favourable with the impact of 0.05% addition on the enteric microorganism quantity of piglet.Illustrate that the action effect of condensation bud pole bacterium may be relevant with its using dosage.Studies have reported that, the mechanism of action of condensation bud pole bacterium, mainly by promoting propagation and the growth of profitable strain (bifidus bacillus, milk-acid bacteria, bacillus faecalis etc.) in enteron aisle, prevents pathogenic bacteria and the harmful bacteria abnormality proliferation in enteron aisle from playing a role.
Table 4 genus bacillus BFS-2 is on the impact of microbe population in weanling pig ight soil
3 conclusions
Equally, adopt the method for examples of implementation 2 and 3 to obtain condensation bud pole bacterium reagent and test with reference to the method for examples of implementation 4, except viable count difference, also obtain similar result (specific experiment slightly).As can be seen here, add in weanling pig daily ration concentration be 0.05% and 0.1% condensation bud pole bacterium can significantly improve average daily gain and the feed efficiency of piglet; Improve quantity of useful microbe in ight soil simultaneously, reduce pernicious bacteria quantity, improve the immunity function of weanling pig.
Claims (5)
1. for a Bacillus coagulans preparation for livestock and poultry cultivation, wherein, said preparation comprises and is deposited at Wuhan, China, and China typical culture collection center preservation, deposit number is the BFS-2 bacterial strain of CCTCCNo:M2011422.
2. for a preparation method for the Bacillus coagulans preparation of livestock and poultry cultivation, the method obtains according to the following steps: with potato glucose solid medium for slant medium; Take potato dextrose broth as shaking flask bacterium culture medium; Zulkovsky starch 2-5% by weight, yeast extract paste 0.5-1%, peptone 0.5-1.5%, dregs of beans 1.0-1.5%, water complements to 100% and is mixed with seeding tank and ferment tank substratum; The Bacillus coagulans bacterial classification wherein adopted is for being deposited at Wuhan, China, and China typical culture collection center preservation, deposit number is the BFS-2 bacterial strain of CCTCCNo:M2011422, and is prepared from after fermentation at different levels, and wherein viable count is 1X10
8-5X10
8the Bacillus coagulans of CFU/g.
3. prepare a method according to claim 2, it is characterized in that carrying out according to the following steps:
(1) preparation of substratum at different levels, wherein:
A) slant medium: potato glucose solid medium, for subsequent use;
B) shaking flask bacterium culture medium: potato dextrose broth, for subsequent use;
C) seeding tank fermention medium: Zulkovsky starch 2-5% by weight, yeast extract paste 0.5-1%, peptone 0.5-1.5%, dregs of beans 1.0-1.5%, water complements to 100%;
D) fermentation tank culture medium: Zulkovsky starch 2-5% by weight, yeast extract paste 0.5-1%, peptone 0.5-1.5%, dregs of beans 1.0-1.5%, water complements to 100%;
(2) cultivation of slant strains: by preservation of bacteria strain: CGMCCNo.2288 Bacillus coagulans BFS-2 inoculation is on step (1) slant medium, cultivate 18-24 hours in the incubator of 28-37 DEG C, directly apply or put into refrigerator and cooled after taking-up to hide, for subsequent use;
(3) shaking flask bacterial classification amplification: under aseptic technique, gets in ring slant strains access 60ml step (1) shaking flask bacterium culture medium, in 28-37 DEG C, in the shaking table of 110-220 revs/min, cultivate 18-24 hours, take out and directly apply or put into refrigerator and cooled Tibetan, for subsequent use;
(4) seeding tank fermentation: step (1) seeding tank fermention medium is put into seeding tank, in seeding tank, temperature 121 DEG C kept after 30 minutes, and pressurize, to 0.1MPa, is cooled to 28-37 DEG C after carrying out steam high-temperature sterilization; Under flame protection, access step (3) shaking flask bacterial classification rapidly by inoculation valve in seeding tank, inoculum size is 5-10%; Start stirrer rotating speed 110-220 revs/min, temperature 28-37 DEG C, air flow 4m
3/ h, tank pressure 0.02-0.06MPa, pH nature, cultivates 18-24 hours, for subsequent use;
(5) preparation of bulk article: step (1) ferment tank substratum is put into fermentor tank, temperature 121 DEG C in fermentor tank, pressurize, to 0.1MPa, keeps, after 30 minutes, substratum is cooled to 28-37 DEG C; In fermentor tank, access step (4) seeding tank bacterial classification by culture transferring pipeline, inoculum size is 5-10%; Start stirrer rotating speed 110-220 revs/min, temperature 28-37 DEG C, air flow 20m
3/ h, tank pressure 0.02-0.06MPa, pH nature, cultivate 18-24 hours, inspection by sampling, carry out aftertreatment (product form needs to be produced into solid or liquid according to client) according to target level of product quality, filling, sealing, for the microbial preparation of livestock and poultry cultivation.
4. one kind comprises the feed of the Bacillus coagulans preparation prepared according to Claims 2 or 3, Bacillus coagulans preparation described in consumption wherein by mass percentage for 0.005-0.1% in feed per ton adds, Direct-fed after stirring, wherein, said preparation comprises and is deposited at Wuhan, China, China typical culture collection center preservation, deposit number is the BFS-2 bacterial strain of CCTCCNo:M2011422 or the cultured products of this bacterial strain, and wherein viable count is 1X10
8-5X10
8the Bacillus coagulans of CFU/g.
5. feed according to claim 4, the mass percent of Bacillus coagulans is 0.05%.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107686826A (en) * | 2017-11-08 | 2018-02-13 | 武汉轻工大学 | The preparation method and pig feed of a kind of bacillus coagulans and its fermentation process, bacillus coagulans preparation |
| CN107712353A (en) * | 2017-11-10 | 2018-02-23 | 王芳 | A kind of feed addictive for preventing diarrhea of pigs and preparation method and application |
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| CN110396480A (en) * | 2018-04-24 | 2019-11-01 | 广东怡和科洁科技有限公司 | A kind of bacillus coagulans XP and its application in Feed Manufacturing |
| CN113604404A (en) * | 2021-08-31 | 2021-11-05 | 四川润格生物科技有限公司 | Bacillus coagulans YSF17 and application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107686826A (en) * | 2017-11-08 | 2018-02-13 | 武汉轻工大学 | The preparation method and pig feed of a kind of bacillus coagulans and its fermentation process, bacillus coagulans preparation |
| CN107686826B (en) * | 2017-11-08 | 2022-07-12 | 武汉轻工大学 | Bacillus coagulans and fermentation method thereof, preparation method of bacillus coagulans preparation and pig feed |
| CN107712353A (en) * | 2017-11-10 | 2018-02-23 | 王芳 | A kind of feed addictive for preventing diarrhea of pigs and preparation method and application |
| CN108703253A (en) * | 2018-04-09 | 2018-10-26 | 华中农业大学 | A kind of feeding natural plants fermentation preparation and its preparation method and application preventing livestock and poultry |
| CN110396480A (en) * | 2018-04-24 | 2019-11-01 | 广东怡和科洁科技有限公司 | A kind of bacillus coagulans XP and its application in Feed Manufacturing |
| CN113604404A (en) * | 2021-08-31 | 2021-11-05 | 四川润格生物科技有限公司 | Bacillus coagulans YSF17 and application thereof |
| CN113604404B (en) * | 2021-08-31 | 2022-08-16 | 四川润格生物科技有限公司 | Bacillus coagulans YSF17 and application thereof |
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