CN109874920A - A kind of composite microbial feed additive and preparation method thereof - Google Patents
A kind of composite microbial feed additive and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of composite microbial feed additives, are prepared by the raw material of following parts by weight: 30-50 parts of streptococcus acidi lactici fermented solution, 10-15 parts of bacillusmusilaginosiengineering fermentation liquid, 10-15 parts of long shoot trichoderma fermentation liquid, 20-30 parts of chlorella algae solution.The present invention also provides a kind of preparation methods of composite microbial feed additive.Composite microbial feed additive provided by the invention can improve intestines function of piglings, improve weanling pig adding weight and feed efficiency, also has the function of significant antibacterial, antiviral, the immunity of piglet can significantly be improved, there is the phenomenon that diarrhea after effective prevention weaned piglet, has wide application prospect.
Description
Technical field
The invention belongs to animal feed processing technique fields, and in particular to a kind of composite microbial feed additive and its system
Preparation Method.
Background technique
Additive for microbe feedstuff is a kind of special feed addictive, is made by the physiological metabolism of microbial strains
With the growth for promoting cultivated animals, the use of drug is reduced, improves the quality of cultivated animals, realizes green cultivation.Due to micro- life
Object feed addictive has so many excellent characteristics, and more and more researchers have had begun the research of this respect, especially
Being microorganisms feed addictive has become current research hotspot in the application of pig industry.
In pig industry, epidemic disease is propagated in order to reduce sow to piglet, wean is carried out to the piglet of 3-5 week old, still,
Piglet after wean often occurs diarrhea, will affect piglet in this way since autodigestion function is not perfect, hypoimmunity
Normal growth and development, also results in piglet death if serious, this situation restricts early weaning piglet technology
It promotes and applies.
It is to add antibiotic in weanling pig feed to improve piglet for major measure that this situation is taken at present
Immunity, prevention of diarrhea in piglets promote growth, but antibiotic is used for a long time and is also easy to produce drug tolerant bacteria, these bacteriums are easy logical
Crossing food chain influences human health.Therefore, it is necessary to which additive for microbe feedstuff to be applied to the breeding process of weanling pig
In, to solve the problems, such as that current weanling pig is easy diarrhea.
Summary of the invention
The present invention provides a kind of composite microbial feed additive, solves and improved in the prior art using antibiotic
Pigling immunity, prevention of diarrhea in piglets are also easy to produce drug tolerant bacteria, and drug tolerant bacteria is easy to influence human health by food chain
The problem of.
The first purpose of the invention is to provide a kind of composite microbial feed additives, by the raw material system of following parts by weight
It is standby to form: 30-50 parts of streptococcus acidi lactici fermented solution, 10-15 parts of bacillusmusilaginosiengineering fermentation liquid, 10-15 parts of long shoot trichoderma fermentation liquid,
20-30 parts of chlorella algae solution.
Wherein, the streptococcus acidi lactici fermented solution, in the bacillusmusilaginosiengineering fermentation liquid living bacteria count be 1.0 ×
1010-3.0×1010CFU/mL；The long shoot trichoderma fermentation liquid miospore amount is 1.0 × 108-1.5×109A/mL；It is described small
The concentration of ball algae algae solution is 1.0 × 105-2.0×105A/mL.
A second object of the present invention is to provide a kind of preparation methods of composite microbial feed additive, including following step
It is rapid:
Step 1, lactobacillus solution, bacillusmusilaginosiengineering seed liquor are prepared respectively；
Step 2, streptococcus acidi lactici fermented solution is prepared
Lactobacillus solution is inoculated in YPD fluid nutrient medium, then with the oscillation speed of 200r/min at 35 ± 2 DEG C
Degree culture to late log phase, culture finishes to obtain streptococcus acidi lactici fermented solution；
Step 3, bacillusmusilaginosiengineering fermentation liquid is prepared
It will pulverize after Pericarpium Kaki drying, obtain Pericarpium Kaki powder；
It will pulverize after the drying of persimmon core, obtain persimmon core powder；
Pericarpium Kaki powder, persimmon core powder, beef extract-peptone fluid nutrient medium are mixed according to the mass ratio of 1-2:2-5:20
Uniformly, bacillus mucilaginosus medium matter is obtained；
It is inoculated with bacillusmusilaginosiengineering seed liquor into bacillus mucilaginosus medium matter, is uniformly mixed, then 28
With the hunting speed culture of 200r/min to late log phase at ± 2 DEG C, bacillusmusilaginosiengineering bacterium germination liquid is obtained；
Step 4, long shoot trichoderma fermentation liquid is prepared
Long shoot trichoderma is inoculated in PDB fluid nutrient medium according to the inoculum concentration that mass fraction is 5%, is trained in 30 ± 2 DEG C
5d is supported, long shoot trichoderma fermentation material is obtained；Then it with the long shoot trichoderma spore on distillation water elution long shoot trichoderma fermentation material, is grown
Branch trichoderma fermentation liquid；
Step 5, chlorella algae solution is prepared
By chlorella according to mass fraction be 5% inoculum concentration be inoculated in BG11 culture medium, 25 ± 1 DEG C, illumination it is strong
Culture obtains chlorella algae solution to logarithmic growth phase under conditions of spending 5000lux, Light To Dark Ratio 12h:12h；
Step 6,30-50 parts of streptococcus acidi lactici fermented solution, 10-15 parts of bacillusmusilaginosiengineering fermentation liquid, length are weighed by weight
Branch 10-15 parts of trichoderma fermentation liquid, 20-30 parts of chlorella algae solution, are uniformly mixed, obtain mixed fermentation liquid, mixed fermentation liquid is existed
Aeration-drying arrives the composite microbial feed additive to moisture content≤10% under the conditions of 35-40 DEG C.
Preferably, the lactobacillus solution, the bacillusmusilaginosiengineering seed liquor inoculum concentration be 10ml seed
Liquid/kilogram matrix.
Preferably, the granularity of the Pericarpium Kaki powder and the persimmon core powder is 10-20 mesh.
Compared with prior art, the beneficial effects of the present invention are:
1) present invention is used cooperatively using multiple-microorganism, plays effect using its synergistic function；Wherein, lactic acid bacteria
It can promote absorption of the piglet to nutriment in feed, also have and increase intestine beneficial bacteria colony, improve piglet gastrointestinal function
The effect of；Bacillusmusilaginosiengineering can be generated in intestine of young pigs a variety of hormonal substances, biological enzyme, glycosaminoglycan, protein,
Amino acids substance can promote piglet development, enhance piglet disease-resistant poison ability；Long shoot trichoderma can use pathogen bacterium
Filament is that nutrition is grown, bred, while also be can induce and itself being generated a variety of extracellular cytohydrolists, molten by coordinated enzyme
Effect makes pathogen cell disruption, and can generate stronger antibacterial substance, produces to growth of pathogenic bacteria in intestine of young pigs
Raw inhibiting effect；Chlorella is full of nutrition, contains natural protein, amino acid, unsaturated fatty acid, vitamin, minerals
Deng, full nutrition can be provided for piglet for its growth, and chlorella also certain antibacterial action, bacillusmusilaginosiengineering,
Long shoot trichoderma, chlorella number of mechanisms collective effect prevent the growth of pathogen in intestine of young pigs, breeding, enhance the anti-of piglet
Sick effect.
2) composite microbial feed additive provided by the invention has the function of significant antibacterial, antiviral, Ke Yixian
, there is the phenomenon that diarrhea after effectively preventing weaned piglet in the immunity of the raising piglet of work；Intestines function of piglings can also be improved,
Weanling pig adding weight and feed efficiency are improved, has broad application prospect.
Specific embodiment
In order to enable those skilled in the art to more fully understand, technical solution of the present invention is practiced, below with reference to specific
The invention will be further described for embodiment, but illustrated embodiment is not as a limitation of the invention.
Lactic acid bacteria used, bacillusmusilaginosiengineering, long shoot trichoderma bacterium, chlorella are in microbial bacteria in following embodiments
The existing strain that kind preservation administrative center is commercially available, is not related to the exploitation of novel bacterial, pertains only to answering for these types of existing bacterial strain
With.Also, lactic acid bacteria as used in the following examples is specially that China typical culture collection center deposit number is CCTCC
The bacterial strain of NO:M2017382；Bacillusmusilaginosiengineering is specially China Committee for Culture Collection of Microorganisms's common micro-organisms
Center deposit number is the bacterial strain of CGMCC No.16005；Long shoot trichoderma bacterium is specially Chinese microorganism strain preservation conservator
The bacterial strain that meeting common micro-organisms center deposit number is CGMCC No.8723；Chlorella is preserved in China typical culture collection
Center, deposit number are CCTCC NO:M209256；
Lactobacillus solution, bacillusmusilaginosiengineering seed liquor are obtained using conventional method culture in embodiment,
Test method described in following each embodiments is unless otherwise specified conventional method.
Embodiment 1
A kind of composite microbial feed additive is prepared by the raw material of following parts by weight: 30 parts of streptococcus acidi lactici fermented solution,
15 parts of bacillusmusilaginosiengineering fermentation liquid, 10 parts of long shoot trichoderma fermentation liquid, 20 parts of chlorella algae solution；
Wherein, streptococcus acidi lactici fermented solution, living bacteria count is 2.6 × 10 in bacillusmusilaginosiengineering fermentation liquid10CFU/mL；
Long shoot trichoderma fermentation liquid miospore amount is 1.0 × 108A/mL；The concentration of chlorella algae solution is 1.8 × 105A/mL.
It is specific the preparation method is as follows:
Step 1, lactobacillus solution, bacillusmusilaginosiengineering seed liquor are prepared respectively；
Step 2, streptococcus acidi lactici fermented solution is prepared
Lactobacillus solution is inoculated in YPD fluid nutrient medium, then with the oscillation speed of 200r/min at 35 ± 2 DEG C
Degree culture to late log phase, culture finishes to obtain streptococcus acidi lactici fermented solution；
Step 3, bacillusmusilaginosiengineering fermentation liquid is prepared
It will pulverize after Pericarpium Kaki drying, cross 10 meshes, obtain Pericarpium Kaki powder；
It will pulverize after the drying of persimmon core, cross 10 meshes, obtain persimmon core powder；
Pericarpium Kaki powder, persimmon core powder, beef extract-peptone fluid nutrient medium are uniformly mixed according to the mass ratio of 1:2:20,
Obtain bacillus mucilaginosus medium matter；
It is inoculated with bacillusmusilaginosiengineering seed liquor into bacillus mucilaginosus medium matter, is uniformly mixed, then 28
With the hunting speed culture of 200r/min to late log phase at ± 2 DEG C, bacillusmusilaginosiengineering bacterium germination liquid is obtained；
Step 4, long shoot trichoderma fermentation liquid is prepared
Long shoot trichoderma is inoculated in PDB fluid nutrient medium according to the inoculum concentration that mass fraction is 5%, is trained in 30 ± 2 DEG C
5d is supported, long shoot trichoderma fermentation material is obtained；Then it with the long shoot trichoderma spore on distillation water elution long shoot trichoderma fermentation material, is grown
Branch trichoderma fermentation liquid；
Step 5, chlorella algae solution is prepared
By chlorella according to mass fraction be 5% inoculum concentration be inoculated in BG11 culture medium, 25 ± 1 DEG C, illumination it is strong
To logarithmic growth phase, obtaining concentration is 1.8 × 10 for culture under conditions of spending 5000lux, Light To Dark Ratio 12h:12h5The bead of a/mL
Algae algae solution；
Step 6,30-50 parts of streptococcus acidi lactici fermented solution, 10-15 parts of bacillusmusilaginosiengineering fermentation liquid, length are weighed by weight
Branch 10-15 parts of trichoderma fermentation liquid, 20-30 parts of chlorella algae solution, are uniformly mixed, obtain mixed fermentation liquid, mixed fermentation liquid is existed
Under the conditions of 35-40 DEG C aeration-drying to moisture content be 5.3% to get arrive composite microbial feed additive.
Embodiment 2
A kind of composite microbial feed additive is prepared by the raw material of following parts by weight: 40 parts of streptococcus acidi lactici fermented solution,
10 parts of bacillusmusilaginosiengineering fermentation liquid, 12 parts of long shoot trichoderma fermentation liquid, 25 parts of chlorella algae solution；
Wherein, streptococcus acidi lactici fermented solution, living bacteria count is 1.0 × 10 in bacillusmusilaginosiengineering fermentation liquid10CFU/mL；
Long shoot trichoderma fermentation liquid miospore amount is 1.0 × 109A/mL；The concentration of chlorella algae solution is 1.5 × 105A/mL.
It is specific that the preparation method is the same as that of Example 1, the difference is that change the formula of embodiment 1 into embodiment 2, meanwhile,
Bacillusmusilaginosiengineering fermentation liquid the preparation method is as follows:
It will pulverize after Pericarpium Kaki drying, cross 20 meshes, obtain Pericarpium Kaki powder；
It will pulverize after the drying of persimmon core, cross 20 meshes, obtain persimmon core powder；
Pericarpium Kaki powder, persimmon core powder, beef extract-peptone fluid nutrient medium are uniformly mixed according to the mass ratio of 1:5:20,
Obtain bacillus mucilaginosus medium matter；
It is inoculated with bacillusmusilaginosiengineering seed liquor into bacillus mucilaginosus medium matter, is uniformly mixed, then 28
With the hunting speed culture of 200r/min to late log phase at ± 2 DEG C, bacillusmusilaginosiengineering bacterium germination liquid is obtained.Embodiment 2 is made
The moisture content of the composite microbial feed additive obtained is 10%.
Embodiment 3
A kind of composite microbial feed additive is prepared by the raw material of following parts by weight: 50 parts of streptococcus acidi lactici fermented solution,
12 parts of bacillusmusilaginosiengineering fermentation liquid, 15 parts of long shoot trichoderma fermentation liquid, 30 parts of chlorella algae solution；
Wherein, streptococcus acidi lactici fermented solution, living bacteria count is 3.0 × 10 in bacillusmusilaginosiengineering fermentation liquid10CFU/mL；
Long shoot trichoderma fermentation liquid miospore amount is 1.5 × 109A/mL；The concentration of chlorella algae solution is 1.0 × 105A/mL.
It is specific that the preparation method is the same as that of Example 1, the difference is that change the formula of embodiment 1 into embodiment 3, meanwhile,
Bacillusmusilaginosiengineering fermentation liquid the preparation method is as follows:
It will pulverize after Pericarpium Kaki drying, cross 20 meshes, obtain Pericarpium Kaki powder；
It will pulverize after the drying of persimmon core, cross 20 meshes, obtain persimmon core powder；
Pericarpium Kaki powder, persimmon core powder, beef extract-peptone fluid nutrient medium are uniformly mixed according to the mass ratio of 2:2:20,
Obtain bacillus mucilaginosus medium matter；
It is inoculated with bacillusmusilaginosiengineering seed liquor into bacillus mucilaginosus medium matter, is uniformly mixed, then 28
With the hunting speed culture of 200r/min to late log phase at ± 2 DEG C, bacillusmusilaginosiengineering bacterium germination liquid is obtained.Embodiment 3 is made
The moisture content of the composite microbial feed additive obtained is 3.8%.
Embodiment 4
A kind of composite microbial feed additive is prepared by the raw material of following parts by weight: 30 parts of streptococcus acidi lactici fermented solution,
15 parts of bacillusmusilaginosiengineering fermentation liquid, 10 parts of long shoot trichoderma fermentation liquid, 20 parts of chlorella algae solution；
Wherein, streptococcus acidi lactici fermented solution, living bacteria count is 1.8 × 10 in bacillusmusilaginosiengineering fermentation liquid10CFU/mL；
Long shoot trichoderma fermentation liquid miospore amount is 3.5 × 108A/mL；The concentration of chlorella algae solution is 2.0 × 105A/mL.
It is specific that the preparation method is the same as that of Example 1, the difference is that bacillusmusilaginosiengineering fermentation liquid the preparation method is as follows:
It will pulverize after Pericarpium Kaki drying, cross 10 meshes, obtain Pericarpium Kaki powder；
It will pulverize after the drying of persimmon core, cross 10 meshes, obtain persimmon core powder；
Pericarpium Kaki powder, persimmon core powder, beef extract-peptone fluid nutrient medium are uniformly mixed according to the mass ratio of 2:5:20,
Obtain bacillus mucilaginosus medium matter；
It is inoculated with bacillusmusilaginosiengineering seed liquor into bacillus mucilaginosus medium matter, is uniformly mixed, then 28
With the hunting speed culture of 200r/min to late log phase at ± 2 DEG C, bacillusmusilaginosiengineering bacterium germination liquid is obtained.Embodiment 4 is made
The moisture content of the composite microbial feed additive obtained is 8.7%.
In order to verify the effect of Pericarpium Kaki powder and persimmon core powder, the present invention is provided with following comparative examples.
Comparative example 1
Bacillusmusilaginosiengineering fermentation liquid the preparation method is as follows:
It will pulverize after Pericarpium Kaki drying, cross 10 meshes, obtain Pericarpium Kaki powder；
Pericarpium Kaki powder is uniformly mixed with beef extract-peptone fluid nutrient medium according to the mass ratio of 1:20, gel-shaped is obtained
Bacillus culture medium matter；
It is inoculated with bacillusmusilaginosiengineering seed liquor into bacillus mucilaginosus medium matter, is uniformly mixed, then 28
With the hunting speed culture of 200r/min to late log phase at ± 2 DEG C, bacillusmusilaginosiengineering bacterium germination liquid is obtained.
Comparative example 2
Bacillusmusilaginosiengineering fermentation liquid the preparation method is as follows:
It will pulverize after the drying of persimmon core, cross 10 meshes, obtain persimmon core powder；
Persimmon core powder is uniformly mixed with beef extract-peptone fluid nutrient medium according to the mass ratio of 2:20, gel-shaped is obtained
Bacillus culture medium matter；
It is inoculated with bacillusmusilaginosiengineering seed liquor into bacillus mucilaginosus medium matter, is uniformly mixed, then 28
With the hunting speed culture of 200r/min to late log phase at ± 2 DEG C, bacillusmusilaginosiengineering bacterium germination liquid is obtained.
Comparative example 3
Bacillusmusilaginosiengineering fermentation liquid the preparation method is as follows:
It is inoculated with bacillusmusilaginosiengineering seed liquor into beef extract-peptone fluid nutrient medium, is uniformly mixed, then 28
With the hunting speed culture of 200r/min to late log phase at ± 2 DEG C, bacillusmusilaginosiengineering bacterium germination liquid is obtained.
Detect bacillusmusilaginosiengineering in bacillusmusilaginosiengineering bacterium germination liquid obtained in embodiment 1 and comparative example 1-3
Living bacteria count is specifically shown in Table 1.
Living bacteria count in 1 bacillusmusilaginosiengineering fermentation liquid of table
| Method | Embodiment 1 | Comparative example 1 | Comparative example 2 | Comparative example 3 |
| Living bacteria count (1010CFU/ml) | 2.6 | 1.2 | 1.8 | 0.5 |
As shown in Table 1, the living bacteria count of bacillusmusilaginosiengineering is most in the bacillusmusilaginosiengineering bacterium germination liquid of comparative example 3
Low, other groups are higher, illustrate that Pericarpium Kaki powder and persimmon core powder are added in fermentation medium to be helped to increase gel-shaped gemma bar
The growth of bacterium, and Pericarpium Kaki powder and persimmon core powder are free of in common beef extract-peptone fluid nutrient medium, therefore living bacteria count
It is lower；
Pericarpium Kaki powder and persimmon core powder are all added in embodiment 1, comparative example 1-2, therefore living bacteria count is higher, than independent
It is good using beef extract-peptone fluid nutrient medium effect, illustrate Pericarpium Kaki powder and persimmon core powder for promoting bacillusmusilaginosiengineering
Growth play a significant role；By comparing embodiment 1, the data of comparative example 1 and comparative example 2, it has been found that persimmon core powder
Effect is good compared with Pericarpium Kaki powder, and the effect that Pericarpium Kaki powder and persimmon core powder are used cooperatively is more preferable, illustrates that the two can be synergistic.
The above test results show that the culture medium culture of the embodiment of the present invention has and dramatically increases bacillusmusilaginosiengineering and have
Imitate the effect of viable count.
Effect in order to further illustrate the present invention, the present invention is also provided with following comparative examples, specific as follows:
Comparative example 4
Streptococcus acidi lactici fermented solution is contained only in additive for microbe feedstuff.
Comparative example 5
A kind of composite microbial feed additive, formula and embodiment 1 are identical, the difference is that comparative example 1 is matched
The preparation of bacillusmusilaginosiengineering fermentation liquid is also free of without addition bacillusmusilaginosiengineering fermentation liquid in side, and in preparation method
Step.
Comparative example 6
A kind of composite microbial feed additive, formula and embodiment 1 are identical, the difference is that comparative example 1 is matched
The preparation step of long shoot trichoderma fermentation liquid is also free of without addition long shoot trichoderma fermentation liquid in side, and in preparation method.
Comparative example 7
A kind of composite microbial feed additive, formula and embodiment 1 are identical, the difference is that comparative example 1 is matched
The preparation step of chlorella algae solution is also free of without addition chlorella algae solution in side, and in preparation method.
It should be noted that in the embodiment of the present invention and comparative example, lactobacillus solution, bacillusmusilaginosiengineering seed liquor
Inoculum concentration be 10ml seed liquor/kilogram matrix.
It is only tested for the property below with the additive for microbe feedstuff that embodiment 1 and comparative example 4-7 are prepared, to this
The effect of invention is illustrated.
1, subjects
Experimental animal chooses age in days 28 ± 1, average weight 5.5-6.5kg, and the health raised under identical rearing conditions
Piglet 120,6 groups are randomly divided into, every group 20, group difference is not significant.Test group is 1 group, and feeding embodiment 1 is prepared micro-
Biological feed additive+normal diet；Control group is 4 groups, the additive for microbe feedstuff that respectively prepared by feeding comparative example 1-4+
Normal diet；Drug control group feeds normal diet+antibiotic (sulfate colistin 10mg/kg+ benefit proceomycin 25mg/kg).Institute
There are the sheep red blood cell (SRBC) suspension that piglet is 1.0% through ear vein injection 5mL mass concentration on the day of 28 ages in days, experimental period 14d.
It should be noted that in above-mentioned each group the additive amount of additive for microbe feedstuff and normal diet additive amount quality
Than for 1:99, the mass ratio of antibiotic and normal diet is also 1:99, and normal diet is purchase from the emerging Anhui livestock technology in Anhui
10% piglet compound premixed feed of development corporation, Ltd..
2, test method
Each group piglet is raised using high bed crack, conventionally carries out disinfection to pig house, it is freely eaten, drink
Water.Before the test, after testing 7d, the early morning after test 14d, under the conditions of empty stomach, take a blood sample respectively from 9 groups of living bodies through ear vein few
Amount then carries out neutrophil leucocyte and peripheral blood lymphocytes percentage through jugular vein blood collection 8-10mL (adding 1% heparin sodium anticoagulant)
Monocyte in the several measurement of measurement, sheep red blood cell (SRBC) antibody titer, the measurement of gamma globulin content and peripheral blood
The measurement of proliferation rate.
Wherein, the measurement of neutrophil leucocyte and peripheral blood lymphocytes percentage carries out the white thin of routine using ear vein blood
Born of the same parents' differential counting；
The measurement of sheep red blood cell (SRBC) antibody titer measures piglet sheep using the conventional micromethod hemagglutination test of ear vein blood
Erythrocyte antibody (EA) potency (sheep red blood cell (SRBC) concentration is 1.0%)；
The measuring method of gamma globulin content is as follows: ear vein blood system is from serum, cellulose acetate membrane electrophoresis method point
From protein component each in serum, OD value is measured in 620nm on spectrophotometer after dyeing, elution, gamma globulin is surveyed and contains relatively
Amount；
The measuring method of mononuclear cell proliferation rate is as follows in peripheral blood: after the dilution of heparin sodium anticoagulated blood Hank ' s liquid, adding
Enter Ficoll separating liquid (GE Healthcare), 2000r/min horizontal centrifugal 30min；Mononuclearcell boundary layer is drawn,
Hank ' s liquid washs 2 times, with 1640 complete culture solution of RPMI adjustment cell concentration for 2 × 10640 hole steril cells are added in/mL
Culture plate, cell suspension 0.1mL is added in every hole, and the RPMI1640 complete culture solution (final concentration for containing or not contain PHA is added
10 μ g/mL), add 3 holes to make stimulation conversion or spontaneous nuclear transformation control, every part of sample.Set 37 DEG C, 5%CO2Incubator culture 66h,
Then MTT liquid (PBS is prepared, pH 7.2), which is added, makes its final concentration of 0.5mg/mL.Continue to cultivate 6h, adds 20%SDS liquid
(contain 50% dimethylformamide, pH 4.7), 37 DEG C of effect 12h, in survey OD value at 570nm on spectrophotometer.
During test, the diarrhea rate of daily every group of piglet in experimental period is recorded, and in the 14d morning of experimental period to son
Pig carries out empty stomach weighing, calculates pig weight gain and feed-weight ratio.
3, data processing
Statistical disposition is carried out using SPSS12.0 statistical software, measurement data data are to indicate, with multiple sample average ratios
Compared with one-way analysis of variance, enumeration data use x2It examines.
4, test result
Test result is as shown in table 2.
Influence/% of the table 2 to piglet neutrophil leucocyte and peripheral blood lymphocytes percentage
From table 2 it can be seen that the additive for microbe feedstuff that the embodiment of the present invention 1 provides can significantly improve piglet blood
Neutrophil leucocyte in system significantly reduces lymphocyte, and effect and effects of antibiotics are suitable.
Influence of the table 3 to piglet serum sheep red blood cell (SRBC) antibody titer (log2)
From table 3 it can be seen that the feed addictive that the embodiment of the present invention 1 provides can significantly improve the immune response of piglet
Reaction, effect and effects of antibiotics are suitable.
Influence of the table 4 to piglet gamma globulin content (OD value)
From table 4, it can be seen that the feed addictive that the embodiment of the present invention 1 provides can significantly increase pigling immunity,
Effect and effects of antibiotics are suitable.
Influence of the table 5 to mononuclear cell proliferation rate in piglet peripheral blood (OD value)
As can be seen from Table 5, peripheral blood mononuclear cells appreciation rate is significantly reduced using antibiotic, but uses the present invention
Embodiment and comparative example human peripheral blood monocyte appreciation rate do not make significant difference.
Influence of the table 6 to weaned piglets
By table 6, it can be concluded that, after the composite microbial feed additive for feeding embodiment 1, the day of piglet searches for food within 2 weeks
Amount, daily increment and diarrhea control rate are all suitable with drug control group, show composite microbial feed additive provided by the invention
The intestinal growth and health that piglet can be significantly improved, the stress reaction after can alleviating weaned piglet, effectively improve piglet and adopt
Appetite and the scale of construction, the effective solution diarrhea problem of piglet.
The present invention describes preferred embodiment, and once a person skilled in the art knows basic creative general
It reads, then additional changes and modifications may be made to these embodiments.So it includes preferred real that the following claims are intended to be interpreted as
It applies example and falls into all change and modification of the scope of the invention.
Obviously, various changes and modifications can be made to the invention without departing from essence of the invention by those skilled in the art
Mind and range.In this way, if these modifications and changes of the present invention belongs to the range of the claims in the present invention and its equivalent technologies
Within, then the present invention is also intended to include these modifications and variations.
Claims (4)
1. a kind of composite microbial feed additive, which is characterized in that be prepared by the raw material of following parts by weight: lactic acid bacteria hair
30-50 parts of zymotic fluid, 10-15 parts of bacillusmusilaginosiengineering fermentation liquid, 10-15 parts of long shoot trichoderma fermentation liquid, chlorella algae solution 20-30
Part.
Wherein, the streptococcus acidi lactici fermented solution, living bacteria count is 1.0 × 10 in the bacillusmusilaginosiengineering fermentation liquid10-
3.0×1010CFU/mL；The long shoot trichoderma fermentation liquid miospore amount is 1.0 × 108-1.5×109A/mL；The chlorella
The concentration of algae solution is 1.0 × 105-2.0×105A/mL.
2. the preparation method of composite microbial feed additive according to claim 1, which is characterized in that including following step
It is rapid:
Step 1, lactobacillus solution, bacillusmusilaginosiengineering seed liquor are prepared respectively；
Step 2, streptococcus acidi lactici fermented solution is prepared
Lactobacillus solution is inoculated in YPD fluid nutrient medium, is then trained at 35 ± 2 DEG C with the hunting speed of 200r/min
It supports to late log phase, culture finishes to obtain streptococcus acidi lactici fermented solution；
Step 3, bacillusmusilaginosiengineering fermentation liquid is prepared
It will pulverize after Pericarpium Kaki drying, obtain Pericarpium Kaki powder；
It will pulverize after the drying of persimmon core, obtain persimmon core powder；
Pericarpium Kaki powder, persimmon core powder, beef extract-peptone fluid nutrient medium are uniformly mixed according to the mass ratio of 1-2:2-5:20,
Obtain bacillus mucilaginosus medium matter；
It is inoculated with bacillusmusilaginosiengineering seed liquor into bacillus mucilaginosus medium matter, is uniformly mixed, then at 28 ± 2 DEG C
Under with the hunting speed culture of 200r/min to late log phase, obtain bacillusmusilaginosiengineering bacterium germination liquid；
Step 4, long shoot trichoderma fermentation liquid is prepared
Long shoot trichoderma is inoculated in PDB fluid nutrient medium according to the inoculum concentration that mass fraction is 5%, in 30 ± 2 DEG C of culture 5d,
Obtain long shoot trichoderma fermentation material；Then with the long shoot trichoderma spore on distillation water elution long shoot trichoderma fermentation material, long shoot wood is obtained
Mould fermentation liquid；
Step 5, chlorella algae solution is prepared
Chlorella is inoculated in BG11 culture medium according to the inoculum concentration that mass fraction is 5%, in 25 ± 1 DEG C, intensity of illumination
Culture obtains chlorella algae solution to logarithmic growth phase under conditions of 5000lux, Light To Dark Ratio 12h:12h；
Step 6,30-50 parts of streptococcus acidi lactici fermented solution, 10-15 parts of bacillusmusilaginosiengineering fermentation liquid, long shoot wood are weighed by weight
Mould fermentation liquid 10-15 parts, 20-30 parts of chlorella algae solution are uniformly mixed, mixed fermentation liquid are obtained, by mixed fermentation liquid in 35-40
Aeration-drying arrives the composite microbial feed additive to moisture content≤10% under the conditions of DEG C.
3. the preparation method of composite microbial feed additive according to claim 2, which is characterized in that the lactic acid bacteria
Seed liquor, the inoculum concentration of the bacillusmusilaginosiengineering seed liquor are 10ml seed liquor/kilogram matrix.
4. the preparation method of composite microbial feed additive according to claim 2, which is characterized in that the Pericarpium Kaki
The granularity of powder and the persimmon core powder is 10-20 mesh.
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