CN109593666A - A kind of compound micro-ecological preparation and its preparation method and application - Google Patents
A kind of compound micro-ecological preparation and its preparation method and application Download PDFInfo
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Abstract
The present invention relates to a kind of compound micro-ecological preparations and its preparation method and application, belong to probiotics technical field.The preparation method of compound micro-ecological preparation of the invention, the following steps are included: Bacillus subtilis strain is accessed in microbial liquid culture medium, in 32~37 DEG C of aerobic fementation cultures to forming gemma, then access S. cervisiae strain is in 28~32 DEG C of aerobic fementations, when S. cervisiae is in stationary phase, while lactobacillus plantarum strain and enterococcus faecium strain are accessed in 30~37 DEG C of closed standing for fermentation, when lactobacillus plantarum and enterococcus faecium are in stationary phase simultaneously, stop fermentation to get.The preparation method of compound micro-ecological preparation of the invention, multi-cultur es stepwise fermentation using different probiotics using nutrition composition difference nutritional substrate is made full use of, save raw material and the energy；And containing there are many probiotics and metabolite abundant in tunning, each strain ratio is naturally occurred, and clinical effectiveness is more preferable.
Description
Technical field
The present invention relates to a kind of compound micro-ecological preparations and its preparation method and application, belong to probiotics technology neck
Domain.
Background technique
Probiotics can generate a variety of digestive ferments, organic acid, vitamin etc. as a kind of additive for farm animal feed, have
Improve microbial population of animal intestinal tract, improve the effects of enteron aisle digestion and absorption and immune function.The preparation side of common compound micro-ecological preparation
There are mainly two types of methods: one kind is solid state fermentation, and a variety of prebiotic strains such as bacillus subtilis, saccharomycete etc. are mixed, are inoculated in
Solid medium ferments, containing there are many probiotics and metabolite abundant in tunning, but during the fermentation
It is highly prone to the pollution of miscellaneous bacteria and fermentation is caused to fail.Second is that single bacterium kind is carried out liquid state fermentation, after fermentation will not
Congener fermentation liquid cooperates according to a certain percentage, prepares compound micro-ecological preparation, the disadvantage is that fermentation substrate cannot be beneficial
Bacterium makes full use of, and be easy to cause secondary fermentation, situations such as Packaging Bottle spalling occurs；Meanwhile the ratio of each strain is artificial preparation,
It is difficult to the natural growth rhythm of complete complex microorganism.In addition, being combined in proportion Tiny ecosystem system with different probiotics bacterial powder
Agent is also compound micro-ecological preparation common in the market；Compound tense is carried out with probiotic powder, the beneficial metabolic product of probiotics exists
It is destroyed in bacterium powder preparation process, meanwhile, certain probiotics such as lactic acid bacteria, saccharomycete are easy death, preparation in the drying process
Bacterium powder in living bacteria count it is lower, it is poor to eventually lead to preparation function and effect.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of compound micro-ecological preparation, the compound microecological system being prepared
Agent contains there are many probiotics and metabolite abundant, and each strain ratio naturally occurs, and clinical effectiveness is more preferable.
The present invention also provides a kind of compound micro-ecological preparation obtained using above-mentioned preparation method and its applications.
In order to achieve the goal above, technical solution used by the preparation method for compound micro-ecological preparation of the invention
It is:
A kind of preparation method of compound micro-ecological preparation, comprising the following steps: Bacillus subtilis strain is accessed into micro- life
In object fluid nutrient medium, in 32~37 DEG C of aerobic fementation cultures to gemma is formed, then access S. cervisiae strain 28~
32 DEG C of aerobic fementations when S. cervisiae is in stationary phase, while accessing lactobacillus plantarum strain and enterococcus faecium strain exists
30~37 DEG C of closed standing for fermentation, when lactobacillus plantarum and enterococcus faecium are in stationary phase simultaneously, stop fermentation to get.
Strain selected by compound micro-ecological preparation of the invention has following characteristic:
Bacillus subtilis: belong to aerobic-type probiotics, can be bred rapidly under conditions of aerobic, into animal intestinal tract after energy
It rapidly depletes the oxygen in environment and causes anaerobic environment, gastrointestinal tract is created good beneficial to anaerobic bacteria such as lactic acid bacteria, Bifidobacterium
Growing environment, to maintain the superiority of the intestinal floras such as lactic acid bacteria, Bifidobacterium；Various enzymes are generated in fermentation process,
As protease, amylase improve the digestive function of animal, and then improve feed benefit by giving animal supplemented with exogenous digestive ferment
With rate；Metabolite such as subtilin, bacitracin have inhibiting effect to pathogenic microorganism, are conducive to maintain intestinal flora balance；
The generation that ammonia can be reduced improves the environment of growth of animals or poultry；To mycotoxin have certain degradation, can reduce because
Feed is mouldy to be damaged to caused by animal body.
S. cervisiae: facultative anaerobic bacteria can breed rapidly under conditions of aerobic, consume the oxygen in enteron aisle, hair
It waves biology and takes oxygen effect by force, facilitate the formation of enteron aisle anaerobic environment, while generating a large amount of high-quality mycoprotein and being utilized for body；
There can be fermentation under anaerobic, generate a variety of metabolites such as digestive ferment, amino acid, vitamin etc., mentioned for body
For good nutrition.The ingredients such as glucan and mannosan in Saccharomyces cerevisiae cells wall are good immunologic stimulant, energy
The immune function of animal body is enough improved, while it can also be promoted to excrete in conjunction with mycotoxin, mitigates toxin and body is caused
Damage.
Lactobacillus plantarum, enterococcus faecium: belong to anaerobic type bacterium, a variety of organic acids are generated in metabolic process, pass through reduction
Gastrointestinal pH facilitates the activity for activating and improving pepsin；Organic acid and lactein in metabolite is to big
Enterobacteria, salmonella etc. have inhibition or killing effect, are conducive to maintain intestinal flora balance；Stimulate the hair of intestine immunity tissue
It educates, improves intestinal immunity；Lactobacillus plantarum and enterococcus faecium belong to lactic acid bacteria, also have to mycotoxin such as aflatoxin
Certain degradation and suction-operated.
It after above four kinds of probiotics enter in animal body, mutually cooperates with, plays trophism, immunological enhancement, promotion
The effects of intestinal flora balance and degradation mycotoxin, the generation of animal intestinal tract disease is advantageously reduced, the production of animal is improved
Performance.
The preparation method of compound micro-ecological preparation of the invention, when using different prebiotic strain stepwise fermentations, due to difference
Nutrition composition that probiotics utilizes is different, and multi-cultur es stepwise fermentation can make full use of nutritional substrate, save raw material and
The energy；And containing there are many probiotics and metabolite abundant in obtained tunning, each strain ratio is naturally occurred, more
Adduction reason, clinical effectiveness are more preferable；Finally, fermentation processes technology content is higher, it is competing to be conducive to medium-sized and small enterprises' product core
Strive the formation of power.
Fermentation process can carry out in the fermenter, and the volume for putting into the microbial liquid culture medium of fermentor accounts for fermentor
The 40~60% of total volume.After putting into microbial liquid culture medium, fermentor is sterilized 40~60min at 115~125 DEG C,
Then Bacillus subtilis strain is accessed again.The volume of fermentor is 0.5~2m3。
Preferably, the viable count of the Bacillus subtilis strain is (0.5~1.0) × 109CFU/mL, S. cervisiae
The viable count of strain is (0.05~1.0) × 109CFU/mL, the viable count of lactobacillus plantarum strain be (1.0~2.0) ×
109CFU/mL, the viable count of enterococcus faecium strain are (1.0~2.0) × 109CFU/mL；The Bacillus subtilis strain connects
Kind of amount is 0.1~0.5%, and the inoculum concentration of the S. cervisiae strain is 0.1~0.5%, the lactobacillus plantarum strain and
The inoculum concentration of enterococcus faecium strain is 0.1~0.3%.
It is further preferred that the specification of the Bacillus subtilis strain is 1.0 × 109CFU/mL, S. cervisiae bacterium
The specification of kind is 1.0 × 109CFU/mL, the specification of lactobacillus plantarum strain are 1.0 × 109CFU/mL, the rule of enterococcus faecium strain
Lattice are 2.0 × 109CFU/mL。
Preferably, the microbial liquid culture medium is grouped as by the group of following mass fraction: bean cake powder 2~3%, corn
Powder 1.5~2%, wheatfeed 1.5~2%, glucose 0.5~1.5%, ammonium sulfate 0.05~0.1%, magnesium sulfate 0.01~
0.03%, manganese sulfate 0.01~0.03%, soybean oil 0.5~1%, surplus are water.The bean cake powder is 80~120 mesh, corn
Powder is 80~120 mesh, wheatfeed is 80~120 mesh.80~120 mesh referred to 80~120 meshes.
It is further preferred that the microbial liquid culture medium is grouped as by the group of following mass fraction: bean cake powder
2.6%, corn flour 1.7%, wheatfeed 1.7%, glucose 0.6%, ammonium sulfate 0.08%, magnesium sulfate 0.02%, manganese sulfate
0.02%, soybean oil 1%, surplus are water.
Preferably, before bacillus subtilis being accessed microbial liquid culture medium, the pH of microbial liquid culture medium is adjusted
It is 7.0~8.5.
Preferably, the time of the aerobic fementation is 20~30h；The time of the closed standing for fermentation is 20~30h.Into
One step is preferred, and the time of the closed standing for fermentation is 24~30h.
Preferably, the volume of gas and the volume ratio of microbial liquid culture medium being passed through during the aerobic fementation be
(1~3): (0.4~0.6).
Preferably, the pressure of aerobic fementation and closed standing for fermentation is 0.04~0.05MPa.
Preferably, fermentation system is stirred with the revolving speed of 110~150r/min during aerobic fementation.Standing for fermentation
It does not stir in the process, stuffiness.
Volume, the stirring rate for the gas that the time of aerobic fementation, pressure and fermentation process are passed through twice can be identical
It can not be identical.
Technical solution used by compound micro-ecological preparation of the invention are as follows:
A kind of compound micro-ecological preparation that the preparation method of above-mentioned compound micro-ecological preparation obtains.
Compound micro-ecological preparation of the invention be used as feed addictive in terms of using used technical solution are as follows:
A kind of application of above-mentioned compound micro-ecological preparation as feed addictive.
When compound micro-ecological preparation of the invention is added into feed as feed addictive, it can be used for treating domestic birds and animals
Bacterial diarrhea, indigestion improve domestic birds and animals production performance.
Compound micro-ecological preparation of the invention is for the symptoms such as poultry bacterial diarrhea, indigestion, every 1mL composite microbial
State preparation converts 1L water, freely drinks, and is used in conjunction 5 days；For the diseases such as the domestic animals such as piglet bacterial diarrhea, indigestion, every head
4mL compound micro-ecological preparation is drunk daily, is used in conjunction 3 days；When carrying out preventative addition for improving breeding performonce fo animals, often
It every 7 days with 1 time, is used in conjunction every time 7 days, every 1mL compound micro-ecological preparation converts 2L water, freely drinks.
Specific embodiment
Below in conjunction with specific embodiment, further description of the technical solution of the present invention.
Use the gas being passed through in bacillus subtilis and S. cervisiae fermentation process for filtrated air in each embodiment.
Embodiment 1
The preparation method of the compound micro-ecological preparation of the present embodiment, comprising the following steps:
1) in 1.5m3Fermentor in be packed into fermenter volume 50% microbial liquid culture medium, adjust microorganism
The pH of fluid nutrient medium is 8.0, in 125 DEG C of sterilizing 40min, cooling；The microbial liquid culture medium is by following mass fraction
Group be grouped as: bean cake powder 2.6%, corn flour 1.7%, wheatfeed 1.7%, glucose 0.6%, ammonium sulfate 0.08%, sulfuric acid
Magnesium 0.02%, manganese sulfate 0.02%, soybean oil 1%, surplus are water；Used bean cake powder is 120 mesh, corn flour be 100 mesh,
Wheatfeed is 100 mesh；
2) to access Bacillus subtilis strain in 0.5% inoculum concentration microbial liquid culture medium in the fermenter,
It ferments for 24 hours at 35 DEG C, obtains bacillus subtilis fermentation liquor；It is 0.05MPa and continuous that the pressure in tank is kept in fermentation process
It is passed through gas, constantly revolving speed used to be stirred for the blender of 150r/min, the total volume for being passed through gas is tank body volume 2
Times；The viable count of used Bacillus subtilis strain is 1.0 × 109CFU/mL；
3) S. cervisiae bacterium is accessed in the bacillus subtilis fermentation liquor obtained in step 1) with 0.3% inoculum concentration
Kind, in 30 DEG C of fermentation 20h, obtain composite fermentation liquid；In fermentation process keep tank in pressure be 0.05MPa it is not open close enter gas
Body constantly uses revolving speed to be stirred for the blender of 120r/min, and the total volume for being passed through gas is tank body volume；It is used
The viable count of S. cervisiae strain is 1.0 × 109CFU/mL；
4) respectively with 0.1% inoculum concentration simultaneously in the composite fermentation liquid obtained in step 3) access lactobacillus plantarum and
Enterococcus faecium strain, 35 DEG C of fermentation 30h to get；Fermentation process stuffiness is not stirred, and holding fermentation pressure inside the tank is
0.04MPa；The viable count of used lactobacillus plantarum strain is 1.0 × 109The viable count of CFU/mL, enterococcus faecium strain is
2.0×109CFU/mL。
The compound micro-ecological preparation of the present embodiment is made using above-mentioned preparation method.
When the compound micro-ecological preparation of the present embodiment is used as feed addictive, it to be used for poultry bacterial diarrhea, indigestion
Equal diseases, every 1mL compound micro-ecological preparation are converted 1L water, are freely drunk, be used in conjunction 5 days；Not for piglet bacillary diarrhea, digestion
Good equal diseases, every is drunk 4mL compound micro-ecological preparation daily, is used in conjunction 3 days；Prevented for improving breeding performonce fo animals
Property addition when, every 7 days with 1 time, be used in conjunction every time 7 days, every 1mL compound micro-ecological preparation converts 2L water, freely drinks.
Embodiment 2
The preparation method of the compound micro-ecological preparation of the present embodiment, comprising the following steps:
1) in 0.5m3Fermentor in be packed into fermenter volume 60% microbial liquid culture medium, adjust microorganism
The pH of fluid nutrient medium is 7.5, in 115 DEG C of sterilizing 60min, cooling；The microbial liquid culture solution is by following quality percentage
The group of ratio is grouped as: bean cake powder 3%, corn flour 1.5%, wheatfeed 1.5%, glucose 1.5%, ammonium sulfate 0.05%, sulfuric acid
Magnesium 0.01%, manganese sulfate 0.03%, soybean oil 1%, surplus are water；Used bean cake powder is 120 mesh, corn flour be 80 mesh,
Wheatfeed is 80 mesh；
2) to access Bacillus subtilis strain in 0.1% inoculum concentration microbial liquid culture medium in the fermenter,
In 37 DEG C of fermentation 30h, bacillus subtilis fermentation liquor is obtained；It is 0.04MPa and continuous that the pressure in tank is kept in fermentation process
It is passed through gas, constantly revolving speed used to be stirred for the blender of 140r/min, the total volume for being passed through gas is tank body volume 3
Times；The viable count of used Bacillus subtilis strain is 1.0 × 109CFU/mL；
3) S. cervisiae bacterium is accessed in the bacillus subtilis fermentation liquor obtained in step 1) with 0.5% inoculum concentration
Kind, in 28 DEG C of fermentation 30h, obtain composite fermentation liquid；In fermentation process keep tank in pressure be 0.04MPa it is not open close enter gas
Body constantly uses revolving speed to be stirred for the blender of 150r/min, and the total volume for being passed through gas is 3 times of tank body volume；Institute
The viable count of the S. cervisiae strain used is 0.05 × 109CFU/mL；
4) respectively with 0.1% inoculum concentration simultaneously in the composite fermentation liquid obtained in step 3) access lactobacillus plantarum and
Enterococcus faecium strain, 37 DEG C of fermentation 30h to get；Fermentation process stuffiness is not stirred, and holding fermentation pressure inside the tank is
0.04MPa；The viable count of used lactobacillus plantarum strain is 2.0 × 109The viable count of CFU/mL, enterococcus faecium strain is
2.0×109CFU/mL。
The compound micro-ecological preparation of the present embodiment is made using above-mentioned preparation method.
When the compound micro-ecological preparation of the present embodiment is used as feed addictive, it to be used for poultry bacterial diarrhea, indigestion
Equal diseases, every 1mL compound micro-ecological preparation are converted 1L water, are freely drunk, be used in conjunction 5 days；Not for piglet bacillary diarrhea, digestion
Good equal diseases, every is drunk 4mL compound micro-ecological preparation daily, is used in conjunction 3 days；Prevented for improving breeding performonce fo animals
Property addition when, every 7 days with 1 time, be used in conjunction every time 7 days, every 1mL compound micro-ecological preparation converts 2L water, freely drinks.
Embodiment 3
The preparation method of the compound micro-ecological preparation of the present embodiment, comprising the following steps:
1) in 2.0m3Fermentor in be packed into fermenter volume 45% microbial liquid culture medium, adjust microorganism
The pH of fluid nutrient medium is 8.5, in 120 DEG C of sterilizing 50min, cooling；The microbial liquid culture medium is by following mass fraction
Group be grouped as: bean cake powder 2%, corn flour 2%, wheatfeed 2%, glucose 0.5%, ammonium sulfate 0.1%, magnesium sulfate
0.03%, manganese sulfate 0.01%, soybean oil 0.5%, surplus are water；Used bean cake powder is 80 mesh, corn flour be 120 mesh,
Wheatfeed is 120 mesh；
2) to access Bacillus subtilis strain in 0.5% inoculum concentration microbial liquid culture medium in the fermenter,
It ferments for 24 hours at 32 DEG C, obtains bacillus subtilis fermentation liquor；It is 0.05MPa and continuous that the pressure in tank is kept in fermentation process
It is passed through gas, constantly revolving speed used to be stirred for the blender of 130r/min, the total volume for being passed through gas is tank body volume；Institute
The viable count of the Bacillus subtilis strain used is 0.5 × 109CFU/mL；
3) S. cervisiae bacterium is accessed in the bacillus subtilis fermentation liquor obtained in step 1) with 0.1% inoculum concentration
Kind, in 32 DEG C of fermentation 20h, obtain composite fermentation liquid；In fermentation process keep tank in pressure be 0.04MPa it is not open close enter gas
Body constantly uses revolving speed to be stirred for the blender of 110r/min, and the total volume for being passed through gas is 2 times of tank body volume；Institute
The viable count of the S. cervisiae strain used is 1.0 × 109CFU/mL；
4) respectively with 0.3% inoculum concentration simultaneously in the composite fermentation liquid obtained in step 3) access lactobacillus plantarum and
Enterococcus faecium strain, 30 DEG C fermentation for 24 hours to get；Fermentation process stuffiness is not stirred, and holding fermentation pressure inside the tank is
0.05MPa；The viable count of used lactobacillus plantarum strain is 1.0 × 109The viable count of CFU/mL, enterococcus faecium strain is
1.0×109CFU/mL。
The compound micro-ecological preparation of the present embodiment is made using above-mentioned preparation method.
When the compound micro-ecological preparation of the present embodiment is used as feed addictive, it to be used for poultry bacterial diarrhea, indigestion
Equal diseases, every 1mL compound micro-ecological preparation are converted 1L water, are freely drunk, be used in conjunction 5 days；Not for piglet bacillary diarrhea, digestion
Good equal diseases, every is drunk 4mL compound micro-ecological preparation daily, is used in conjunction 3 days；Prevented for improving breeding performonce fo animals
Property addition when, every 7 days with 1 time, be used in conjunction every time 7 days, every 1mL compound micro-ecological preparation converts 2L water, freely drinks.
Embodiment 4
The preparation method of the compound micro-ecological preparation of the present embodiment, comprising the following steps:
1) in 1.0m3Fermentor in be packed into fermenter volume 55% microbial liquid culture medium, adjust microorganism
The pH of fluid nutrient medium is 7.0, in 115 DEG C of sterilizing 50min, cooling；The microbial liquid culture medium is by following mass fraction
Group be grouped as: bean cake powder 2.3%, corn flour 1.8%, wheatfeed 1.8%, glucose 1.0%, ammonium sulfate 0.08%, sulfuric acid
Magnesium 0.02%, manganese sulfate 0.02%, soybean oil 0.8%, surplus are water；Used bean cake powder is 100 mesh, corn flour 100
Mesh, wheatfeed are 120 mesh；
2) to access Bacillus subtilis strain in 0.3% inoculum concentration microbial liquid culture medium in the fermenter,
In 35 DEG C of fermentation 20h, bacillus subtilis fermentation liquor is obtained；It is 0.04MPa and continuous that the pressure in tank is kept in fermentation process
It is passed through gas, constantly revolving speed used to be stirred for the blender of 110r/min, the total volume for being passed through gas is tank body volume 2
Times；The viable count specification of used Bacillus subtilis strain is 0.8 × 109CFU/mL；
3) S. cervisiae bacterium is accessed in the bacillus subtilis fermentation liquor obtained in step 1) with 0.1% inoculum concentration
Kind, it ferments for 24 hours at 30 DEG C, obtains composite fermentation liquid；In fermentation process keep tank in pressure be 0.05MPa it is not open close enter gas
Body constantly uses revolving speed to be stirred for the blender of 130r/min, and the total volume for being passed through gas is tank body volume；It is used
The viable count of S. cervisiae strain is 0.8 × 109CFU/mL；
4) respectively with 0.2% inoculum concentration simultaneously in the composite fermentation liquid obtained in step 3) access lactobacillus plantarum and
Enterococcus faecium strain, 35 DEG C of fermentation 30h to get；Fermentation process stuffiness is not stirred, and holding fermentation pressure inside the tank is
0.05MPa；The viable count of used lactobacillus plantarum strain is 1.5 × 109The viable count of CFU/mL, enterococcus faecium strain is
1.5×109CFU/mL。
The compound micro-ecological preparation of the present embodiment is made using above-mentioned preparation method.
When the compound micro-ecological preparation of the present embodiment is used as feed addictive, it to be used for poultry bacterial diarrhea, indigestion
Equal diseases, every 1mL compound micro-ecological preparation are converted 1L water, are freely drunk, be used in conjunction 5 days；Not for piglet bacillary diarrhea, digestion
Good equal diseases, every is drunk 4mL compound micro-ecological preparation daily, is used in conjunction 3 days；Prevented for improving breeding performonce fo animals
Property addition when, every 7 days with 1 time, be used in conjunction every time 7 days, every 1mL compound micro-ecological preparation converts 2L water, freely drinks.
Comparative example
The microbial liquid culture medium that the compound micro-ecological preparation of comparative example uses with embodiment 1, bacillus subtilis and
The gas being passed through in S. cervisiae fermentation process is filtrated air.
The preparation method of the compound micro-ecological preparation of comparative example, comprising the following steps:
1) preparation is without bacteria fermentation culture medium
Respectively number is 1#~3#, volume is 1.5m3Fermentor in be packed into fermenter volume 50% micro- life
Object fluid nutrient medium, the pH of adjustment microbial liquid culture medium are 8.0, then in 125 DEG C of sterilizing 40min, cooling；
2) fermentation of bacillus subtilis, S. cervisiae fermentation, lactobacillus plantarum and enterococcus faecium fermentation are carried out
A) fermentation of bacillus subtilis
Bacillus subtilis strain is accessed in the microbial liquid culture medium in 1# fermentor with 0.5% inoculum concentration,
It ferments for 24 hours at 35 DEG C, obtains bacillus subtilis fermentation liquor；Tank pressure is kept to be 0.05MPa and be stirred continuously in fermentation process
Under conditions of be passed through gas；The revolving speed of the blender used is stirred as 150r/min, the total volume for being passed through gas is tank body volume 2
Times；The viable count of the strain of used bacillus subtilis is 1.0 × 109CFU/mL；
B) S. cervisiae is fermented
S. cervisiae strain is accessed in the microbial liquid culture medium in 2# fermentor with 0.3% inoculum concentration,
30 DEG C of fermentation 20h, obtain saccharomyces cerevisiae fermented liquid；It is 0.05MPa and in the item being stirred continuously that tank pressure is kept in fermentation process
Gas is passed through under part；The revolving speed of the blender used is stirred as 120r/min, the total volume for the gas being passed through is fermenter volume；
The viable count of used S. cervisiae strain is 1.0 × 109CFU/mL；
C) lactobacillus plantarum and enterococcus faecium fermentation
Lactobacillus plantarum is accessed in the microbial liquid culture medium in 3# fermentor simultaneously with 0.1% inoculum concentration respectively
And enterococcus faecium obtains lactobacillus plantarum and enterococcus faecium composite fermentation liquid then in 35 DEG C of fermentation 30h；In fermentation process not
Stirring, stuffiness and holding tank pressure are 0.04MPa, and the viable count of used lactobacillus plantarum strain is 1.0 × 109CFU/
ML, the viable count of enterococcus faecium strain are 2.0 × 109CFU/mL；
3) with bacillus subtilis in every milliliter of compound micro-ecological preparation in the method for plate culture count measurement embodiment 1, make
Brewer yeast bacterium, lactobacillus plantarum and enterococcus faecium viable count, calculate the ratio of all kinds of viable bacterias；The method of plate culture count is used respectively
The viable count in comparative example in each fermentation liquid is measured, and prepares compound probiotic according to the ratio of all kinds of viable bacterias in embodiment 1
Fermentation liquid to get comparative example compound micro-ecological preparation.
The compound micro-ecological preparation of comparative example, in clinical application, suggested use and dosage is the same as embodiment 1.
Experimental example 1
This experimental example investigates the compound micro-ecological preparation of different embodiments in treatment Broiler chicks enteric flora disturbance and digestion
The bad function and effect for being.
1) experimental material
Experiment reagent: compound micro-ecological preparation made from Examples 1 to 4 and comparative example；
Experimental animal: because 6 age in days broiler chicks of diarrhea occur for indigestion, pathogenetic bacteria infection, by Henan Province's Hebi City
Certain broiler chicken field provides.
2) experimental method
Single factor test completely randomized design is taken in experiment, sets 6 groups, every group of 100 morbidity chickens altogether.Wherein 1~4 point of experimental group
The drinking-water of other compound micro-ecological preparation of the feeding containing Examples 1 to 4；Group 5 is control group, feeding compound microecological system containing comparative example
The drinking-water of agent, application method are identical as group 1~4；Group 6 is blank group, and feeding is free of the drinking-water of any drug.
The feeding method of the compound micro-ecological preparation of Examples 1 to 4 and comparative example: every 1mL compound micro-ecological preparation converts 1L
Water is freely drunk, and is used in conjunction 5 days, clinical therapeutic efficacy is shown in Table 1.
Therapeutic effect after 1 each group medication of table
(note: death rate decreasing value is the absolute value of the difference of the death rate of group 1~5 and the death rate of group 6)
Compound micro-ecological preparation clinical effectiveness: as shown in Table 1, diseased chicken is equal in the recovery rate of medication the 3rd, 5 day in group 1~4
It is apparently higher than the 1st~5 group of general mortality rate than the 6th group reduction by 21,20,20,21,12% respectively in the 6, the 5th day of group 5 and group, the 5th day the
Than the 5th group reduction by 9,8,8,9% respectively of 1~4 group of general mortality rate.The result shows that compound micro-ecological preparation of the invention is being treated
When because of indigestion, pathogenetic bacteria infection diarrhea occurs for broiler chicks, function and effect are ideal.
Experimental example 2
This experimental example is investigated the compound micro-ecological preparation of different embodiments in treatment intestine of young pigs Flora Disturbance and is digested not
Function and effect when good.
1) experimental material
Experimental preparation: compound micro-ecological preparation made from Examples 1 to 4 and comparative example；
Experimental animal: because 2 age in days piglets of diarrhea occur for indigestion, pathogenetic bacteria infection, by Henan Province Tongxu County
Pig farm provides.
2) experimental method
Single factor test completely randomized design is taken in experiment, sets 6 groups, every group of 20 hair disease piglets altogether.Wherein test group 1~4
The compound micro-ecological preparation of Examples 1 to 4 is gavaged respectively；Group 5 is control group, gavages comparative example compound micro-ecological preparation, is used
Method is identical as group 1~4；Group 6 is blank group, does not feed any drug.
The feeding method of the compound micro-ecological preparation of Examples 1 to 4 and comparative example: 4mL/ head/day is used in conjunction 3 days, and clinic is controlled
Therapeutic effect is shown in Table 2.
Therapeutic effect after 2 each group medication of table
(note: death rate decreasing value is the absolute value of the difference of the death rate of 1~5 group of group and the death rate of group 6)
Clinical effectiveness: as shown in Table 2, in group 1~4 diarrhea piglet using after of the invention compound micro-ecological preparation 3 days,
Full recovery is normal, the 5th group of (comparative example preparation) recovery rate 80%, the 6th group of (blank control) recovery rate 70%；Give morbidity piglet
It feeds the present invention and after comparative example compound micro-ecological preparation 3 days, is compared with group 6,1~5 group of the death rate reduces by 20% respectively,
20%, 20%, 20%, 10%, recovery rate is respectively increased 30%, 30%, 30%, 30%, 10%.The result shows that of the invention
For compound micro-ecological preparation when treating piglet because of indigestion, pathogenetic bacteria infection generation diarrhea, function and effect are ideal.
Experimental example 3
The compound micro-ecological preparation that this experimental example investigates different embodiments is carried out for improving Production Performance of Weaning Pigs
Function and effect when preventative addition.
1) experimental material
Experimental preparation: Examples 1 to 4 and the compound micro-ecological preparation of comparative example preparation；
Experimental animal: 24 age in days piglets are provided by Henan Province, pig farm, Yuanyang County.
2) experimental method
Single factor test completely randomized design is taken in experiment, sets 6 groups, every group of 45 weanling pigs altogether.Wherein test group 1~4
The compound micro-ecological preparation of Examples 1 to 4 is added respectively into piglet drinking-water；Group 5 is control group, adds comparative example composite microbial
During state preparation to piglet drinks water；Group 6 is blank group, does not feed any drug.
The feeding method of the compound micro-ecological preparation of Examples 1 to 4 and comparative example: experiment periods 24 days, wean went to child care
It is used in conjunction 7 days, was used in conjunction again every 7 days 7 days after house, every 1mL compound micro-ecological preparation converts 2L water, freely drinks.Clinical effectiveness is shown in Table 3.
To Production Performance of Weaning Pigs effect after 3 each group medication of table
Clinical effectiveness: as shown in Table 3, the average end of group 1~5 has been respectively increased 14.68% than 6 (blank groups) of group again,
10.33%, 12.44%, 12.85%, 4.49%, average total feed intake has been respectively increased 15.83%, 10.15%, 13.21%,
15.50%, 5.90%, average total augment weight has been respectively increased 27.32%, 18.58%, 23.11%, 24.33%, 8.80%, material
Meat ratio is below blank group, tests whole 6 test pig dead 1 of group, other test groups are without injures and deaths.The result shows that this hair
When bright compound micro-ecological preparation carries out preventative addition for improving breeding performonce fo animals, function and effect are significant.
Claims (10)
1. a kind of preparation method of compound micro-ecological preparation, it is characterised in that: the following steps are included: by Bacillus subtilis strain
It accesses in microbial liquid culture medium, in 32~37 DEG C of aerobic fementation cultures to gemma is formed, then accesses S. cervisiae bacterium
Kind when S. cervisiae is in stationary phase, while accessing lactobacillus plantarum strain and dung intestines ball in 28~32 DEG C of aerobic fementations
Bacterium strain stops fermentation when lactobacillus plantarum and enterococcus faecium are in stationary phase simultaneously in 30~37 DEG C of closed standing for fermentation,
To obtain the final product.
2. the preparation method of compound micro-ecological preparation according to claim 1, it is characterised in that: the bacillus subtilis
The viable count of strain is (0.05~1.0) × 109CFU/mL, the viable count of S. cervisiae strain be (0.5~1.0) ×
109CFU/mL, the viable count of lactobacillus plantarum strain are (1.0~2.0) × 109The viable count of CFU/mL, enterococcus faecium strain is
(1.0~2.0) × 109CFU/mL；The inoculum concentration of the Bacillus subtilis strain is 0.1~0.5%, the S. cervisiae
The inoculum concentration of strain is 0.1~0.5%, the inoculum concentration of the lactobacillus plantarum strain and enterococcus faecium strain is 0.1~
0.3%.
3. the preparation method of compound micro-ecological preparation according to claim 1 or 2, it is characterised in that: the microorganism liquid
Body culture medium is grouped as by the group of following mass fraction: bean cake powder 2~3%, corn flour 1.5~2%, wheatfeed 1.5~2%,
Glucose 0.5~1.5%, ammonium sulfate 0.05~0.1%, magnesium sulfate 0.01~0.03%, manganese sulfate 0.01~0.03%, soybean
Oil 0.5~1%, surplus is water.
4. the preparation method of compound micro-ecological preparation according to claim 1 or 2, it is characterised in that: by withered grass gemma
Before bacillus accesses microbial liquid culture medium, the pH of adjustment microbial liquid culture medium is 7.0~8.5.
5. the preparation method of compound micro-ecological preparation according to claim 1 or 2, it is characterised in that: the aerobic fementation
Time be 20~30h；The time of the closed standing for fermentation is 20~30h.
6. the preparation method of compound micro-ecological preparation according to claim 1 or 2, it is characterised in that: the aerobic fementation
The volume for the gas being passed through in the process and the volume ratio of microbial liquid culture medium are (1~3): (0.4~0.6).
7. the preparation method of compound micro-ecological preparation according to claim 1 or 2, it is characterised in that: aerobic fementation and close
The pressure for closing standing for fermentation is 0.04~0.05MPa.
8. the preparation method of compound micro-ecological preparation according to claim 1 or 2, it is characterised in that: aerobic fementation process
In fermentation system is stirred with the revolving speed of 110~150r/min.
9. a kind of compound micro-ecological preparation that the preparation method of compound micro-ecological preparation as described in claim 1 obtains.
10. a kind of application of compound micro-ecological preparation as claimed in claim 9 as feed addictive.
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| CN110628683A (en) * | 2019-10-22 | 2019-12-31 | 江苏恒丰强生物技术有限公司 | Probiotic compound live bacterium preparation for preventing and treating chicken diarrhea and preparation method thereof |
| CN110959752A (en) * | 2019-12-18 | 2020-04-07 | 湖北华扬科技发展有限公司 | High-stability microecological compound preparation and preparation method and application thereof |
| CN113841788A (en) * | 2021-09-26 | 2021-12-28 | 山东迅达康兽药有限公司 | Micro-ecological microcapsule coating preparation and preparation method thereof |
| CN114601882A (en) * | 2020-12-08 | 2022-06-10 | 河南炎黄生物工程有限公司 | Preparation method of Chinese herbal medicine microecological preparation for improving growth performance of fishes |
| CN114908026A (en) * | 2022-06-30 | 2022-08-16 | 哈尔滨韶创生物科技有限公司 | Special liquid composite microecological preparation for rumination and preparation method thereof |
| CN115644304A (en) * | 2022-09-16 | 2023-01-31 | 重庆大学 | Process for preparing fermented feed by fermenting shiitake mushroom residues step by probiotics |
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