CN104017751B - Bacillus coagulans, Lactobacillus plantarum and Lactobacillus casei preparation and its preparation - Google Patents
Bacillus coagulans, Lactobacillus plantarum and Lactobacillus casei preparation and its preparation Download PDFInfo
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Abstract
The invention discloses probiotics of a kind of utilization Bacillus coagulans, Lactobacillus plantarum preparation and preparation method thereof, belong to feed microbe technical field.The probiotics carry out solid fermentation using Bacillus coagulans, Lactobacillus plantarum and lactobacillus casei and form with bean cake, wheat bran, Semen Maydis powder as substrate.The fermentation process of the present invention reduces fermenting and producing cost, processes convenient, pollutes little, and device structure is simple, easy to operate, is a kind of probiotic solid fermentation technology that can be widely applied to Tiny ecosystem field；Macro-molecular protein in bean cake, wheat bran and Semen Maydis powder can be degraded to oligopeptide by the probiotics of the present invention, and the anti-nutrient substance in albumen, antigen protein etc. are thoroughly decomposed, so as to improve the protein quality and feeding value of the fermented products such as bean cake, and in fermented product, contain viable bacteria, animal intestinal microorganism species can effectively be adjusted, improve production performance, improve the utilization rate of feedstuff.
Description
Technical field
The invention belongs to feed microbe technical field, more particularly to one kind are with bean cake, wheat bran, Semen Maydis powder as substrate, profit
The application technology that solid fermentation prepares probiotics is carried out with Bacillus coagulans, Lactobacillus plantarum.
Background technology
More than existing probiotic composition using probiotic bacteria directly with liquid milk and soya-bean milk fermentation, obtain functional clabber
Or sour bean milk, or after being adopted liquid fermentation, directly collect probio thalline, be fabricated to probiotics powder agent, tablet or
Capsule.When using liquid fermentation, substantial amounts of contaminated wastewater environment can be produced, post processing is relatively difficult, machinery equipment operation
It is relatively complicated.As probiotic bacteria can improve host's microecological balance, the viable bacteria system of host health level and health status is improved
Agent and metabolite, therefore, the research to probiotic bacteria is very deeply and detailed-oriented now, and has been widely used for work
Each field such as industry, agricultural, environmental protection, family.
At present, probiotic composition is applied to animal microorganism preparation, adds into animal feed also relatively conventional；Example
Such as, the patent of invention of Publication No. CN103202391A《A kind of effective micro- life of compound lactobacillus for preventing and improving Mammitis of cattle
State preparation》, it is CGMCC by the lactobacillus casei HM-09, preserving number that preserving number is CGMCC No.6736 to disclose a kind of
The compound micro-ecological preparation that the Lactobacillus plantarum HM-10 and zeolite powder of No.6741 is combined by mass fraction, effectively can drop
Low somatic number, prevention reduce recessive Mammitis of cattle sickness rate；The patent of invention of Publication No. CN103243043A《One plant is used for
Improve excellent Bacillus coagulans and its application of inferior piglet cultivation》, a kind of preserving number is disclosed for CGMCC No.6729's
Bacillus coagulans, by using this bacterial strain, can effectively improve the health status of piglet, improving the life that inferior piglet cultivates
Produce performance.The patent of invention of Publication No. CN103421711A《For the cheese breast bar of feed additive probiotic solid fermentation
Bacterium, animal bifidobacteria, Lactobacillus plantarum and bacillus subtilises》Disclose for feed additive probiotic solid fermentation
Lactobacillus casei, animal bifidobacteria, Lactobacillus plantarum and bacillus subtilises, the bacillus subtilises HM-66, cheese breast
Bacillus F-08, Lactobacillus plantarum HM-20 and animal bifidobacteria V9 are probiotic bacteria, are preserved in Chinese microorganism strain preservation pipe
Reason committee General Microbiological Culture collection, preserving number be respectively CGMCCNo.6733, CGMCCNo.6735,
CGMCCNo.6744 and CGMCCNo.5470；By the application of this several bacterial strains can be effectively improved the sour fragrance of feedstuff, color and luster and
The organoleptic features such as mouthfeel.
The content of the invention
Object of the present invention is to provide it is a kind of using Bacillus coagulans (Bacillus coagulans) HM-09 and
The bacterium such as Lactobacillus plantarum (Lactobacillus plantarum) HM-10 carry out being segmented admittedly to bean cake, wheat bran and Semen Maydis powder substrate
The method of state fermentation, the probiotics product for that fermented using the method have special sour fragrance, and the viable count for containing is high,
The degradation rate of bean cake protein is also high, additionally it is possible to adjust animal intestinal microorganism species, promotes intestinal absorption, enhancing human body immunity
Power, improves production performance, improves rotting for the utilization rate of feedstuff, effective suppression cultivation later stage body feces and residual bait material, improves
Environmental health, reduces the generation of ammonia nitrogen and nitrite, reduces whole aquaculture cost.
The present invention is achieved through the following technical solutions：
Probiotics prepared by a kind of utilization Bacillus coagulans, Lactobacillus plantarum, its preparation method comprise the steps：
The preparation of seed liquor：By Bacillus coagulans HM-09 30-38 DEG C in calorstat, 170 turns of shaking table cultures 20-
24h；By Lactobacillus plantarum HM-10 and lactobacillus casei (Lactobacillus casei) HM-09 30-38 DEG C in calorstat
Culture 20-24h；
The seed culture medium of Bacillus coagulans HM-09, Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 is：
Peptone 2-5%, glucose 2-5%, adds water to 100%, pH7.0-7.2；
Fermentation culture conditions：Bacillus coagulans HM-09 seed liquor is added to after being mixed in solid medium and is sent out
Ferment 0-16h, inoculates Lactobacillus plantarum HM-10 seed liquor and continues fermentation 12-36h with lactobacillus casei HM-09 seed liquor, altogether
Fermentation time 12-52h, controls fermentation temperature for 30-38 DEG C in sweat, humidity is 60-80%, fermentation ends its free ammonias
Ground state nitrogen content is in more than 480umol/g；
The solid medium is：25-30% bean cake, 25-30% wheat brans, 2-5% Semen Maydis powder, moisture 40-50%,
Initial pH 6.8-7.2.
3-6% of the addition of the Bacillus coagulans HM-09 seed liquor for solid medium gross weight, plant breast bar
2-4% of the addition of bacterium HM-10 seed liquor for solid medium gross weight, the addition of lactobacillus casei HM-09 seed liquor
For the 4-6% of solid medium gross weight.
Bacillus coagulans HM-09 viable count (1.45-2.98) × 10 in probiotics9CFU/g, Lactobacillus plantarum HM-
10 viable counts (1.55-2.65) × 109CFU/g, lactobacillus casei HM-09 viable counts (3.31-9.98) × 109CFU/g。
Addition of the product of the present invention in feedstuff is 25-60g/kg.
Beneficial effect
The present invention using Bacillus coagulans HM-09 and Lactobacillus plantarum HM-10 etc. be fermenting agent to bean cake, wheat bran and
Semen Maydis powder carries out mixed solid fermentation, reduces fermenting and producing cost, processes convenient, pollutes little, and device structure is simple, easy
Operation, is a kind of probiotic solid fermentation technology that can be widely applied to Tiny ecosystem field；Entered using Bacillus coagulans HM-09
Row prior fermentation, can produce the tart flavour small-molecule substance such as a large amount of lactic acid and acetic acid, and it is peculiar that these small-molecule substances give product
Sour fragrance, color and luster and mouthfeel etc., improve the palatability of product；The present invention with the addition of plant for a period of time afterwards in solid fermentation
Lactobacilluss HM-10 and lactobacillus casei HM-09 carry out mixed fermentation, being capable of efficient utilization lactobacillus casei HM-09 bacterium regulation intestinal
The function that interior colony balance, promotion body are digested and assimilated, while fissional effect can also be promoted using lactobacillus casei,
Produce antibody mediated immunity, enhancing human body immunity；The Lactobacillus plantarum distinctive lactobacillin of energy output in reproductive process, because
This utilizes this characteristic, can suppress rotting for bottom feces and residual bait material well in the cultivation later stage, reduce ammonia nitrogen and nitrite
Generation, a large amount of consumptions for reducing chemical industry degradation elements reduce aquaculture cost；Using the benefit of the method production of segmentation solid fermentation
Contain substantial amounts of viable bacteria in probiotics preparation, therefore, the characteristic of each strain is cooperateed with, animal intestinal microbial bacteria can be effectively adjusted
Group, promotes intestinal absorption, enhancing human body immunity power, improves ablactational baby pig diarrhoea with dislocation of bacillary groups, improves the utilization rate of feedstuff, drop
Low whole aquaculture cost.
Product of the present invention is used for the diarrhea rate more than 30% that pigletss can be effectively reduced during the feeding of early weaned piglets, feedstuff
Conversion ratio improve 25-30%.
Specific embodiment
With reference to example, the present invention is further described, and following examples are illustrative, is not determinate, it is impossible to
Protection scope of the present invention is limited with following examples.
Bacillus coagulans HM-09 provided by the present invention, Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 in
On October 29th, 2012 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is respectively
CGMCC No.6729, CGMCC No.6741 and CGMCC No.6736；Preservation address is：Datun Road, Chaoyang District, Beijing City, China
Institute of microbiology of academy of science, postcode：100101；
Embodiment 1
Probiotics prepared by a kind of utilization Bacillus coagulans, Lactobacillus plantarum, its preparation method comprise the steps：
The preparation of seed liquor：Bacillus coagulans HM-09, Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 are from cold storage
Glycerol tube in be inoculated in the culture medium equipped with liquid seeds, the seed culture condition of Bacillus coagulans HM-09：37
DEG C, 170 turns of shaking table cultures 24h；Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 37 DEG C of culture 24h in calorstat；
Wherein seed culture medium is：Peptone 2%, glucose 2%, adds water to 100%, pH7.0-7.2.
Solid fermentation
Fermentation culture conditions：Bacillus coagulans HM-09 seed liquor is added to after being mixed in solid medium and is sent out
Ferment 12h, inoculates Lactobacillus plantarum HM-10 seed liquor and lactobacillus casei HM-09 seed liquor continues fermentation 24h, ferments altogether
Time 36h, controls fermentation temperature for 37 DEG C in sweat, humidity is 70%, and fermentation ends its free ammonical nitrogen contents is
489umol/g；
The solid medium is：29% bean cake, 29% wheat bran, 2% Semen Maydis powder, moisture 40%, initial pH7.0,
115 DEG C of autoclaving 25min.
The addition of the Bacillus coagulans HM-09 seed liquor for solid medium gross weight 5%, Lactobacillus plantarum
The addition of HM-10 seed liquor for solid medium gross weight 3%, the addition of lactobacillus casei HM-09 seed liquor is solid
The 5% of body culture medium gross weight.
Bacillus coagulans HM-09 viable count 2.98 × 10 in probiotics9CFU/g, Lactobacillus plantarum HM-10 viable counts
2.07×109The viable count 5.98 × 10 of CFU/g, lactobacillus casei HM-099CFU/g。
Addition of the product of the present invention in feedstuff is 45g/kg.
Embodiment 2:
Probiotics prepared by a kind of utilization Bacillus coagulans, Lactobacillus plantarum, its preparation method comprise the steps：
The preparation of seed liquor：By Bacillus coagulans HM-09 37 DEG C in calorstat, 170 turns of shaking table cultures 24h；To plant
Thing lactobacilluss HM-10 and lactobacillus casei HM-09 37 DEG C of culture 24h in calorstat.
Wherein seed culture medium is：Peptone 2%, glucose 2%, adds water to 100%, pH7.0-7.2；
Fermentation culture conditions：Bacillus coagulans HM-09 seed liquor is added to after being mixed in solid medium and is sent out
Ferment 16h, inoculates Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 seed liquor continues fermentation 15h, altogether fermentation time
31h, controls fermentation temperature for 37 DEG C in sweat, humidity is 80%, and fermentation ends its free ammonical nitrogen contents exists
486umol/g；
Solid medium is：29% bean cake, 29% wheat bran, 2% Semen Maydis powder, moisture 40%, initial pH 7.0,115
DEG C autoclaving 25min.
The addition of Bacillus coagulans HM-09 seed liquor for solid medium gross weight 3%, Lactobacillus plantarum HM-
The addition of 10 seed liquor for solid medium gross weight 2%, the addition of lactobacillus casei HM-09 seed liquor are trained for solid
The 6% of foster base gross weight.
Bacillus coagulans HM-09 viable count 1.45 × 10 in probiotics9CFU/g, Lactobacillus plantarum HM-10 viable counts
1.55×109CFU/g, lactobacillus casei HM-09 viable count 9.98 × 109CFU/g。
Addition of the product of the present invention in feedstuff is 30g/kg.
Embodiment 3
Probiotics prepared by a kind of utilization Bacillus coagulans, Lactobacillus plantarum, its preparation method comprise the steps：
The preparation of seed liquor：By Bacillus coagulans HM-09 37 DEG C in calorstat, 170 turns of shaking table cultures 24h；To plant
Thing lactobacilluss HM-10 and lactobacillus casei HM-09 37 DEG C of culture 24h in calorstat.
The seed culture medium of Bacillus coagulans HM-09, Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 is：Albumen
Peptone 2%, glucose 2%, adds water to 100%, pH7.0-7.2；
Fermentation culture conditions：Bacillus coagulans HM-09 seed liquor is added to after being mixed in solid medium and is sent out
Ferment 6h, inoculates lactobacillus casei HM-09 and Lactobacillus plantarum HM-10 seed liquor continues fermentation 36h, altogether fermentation time 42h,
Fermentation temperature is controlled in sweat for 37 DEG C, humidity is 60%, fermentation ends its free ammonical nitrogen contents is in 460umol/
g；
Solid medium is：29% bean cake, 29% wheat bran, 2% Semen Maydis powder, moisture 40%, initial pH 7.0,115
DEG C autoclaving 25min.
The addition of Bacillus coagulans HM-09 seed liquor for solid medium gross weight 6%, Lactobacillus plantarum HM-
10 seed liquor addition for solid medium gross weight 4%, the addition of lactobacillus casei HM-09 seed liquor are trained for solid
The 4% of foster base gross weight.
Bacillus coagulans HM-09 viable count 3.18 × 10 in probiotics9CFU/g, Lactobacillus plantarum HM-10 viable counts
2.65×109CFU/g, lactobacillus casei HM-09 viable count 3.31 × 109CFU/g。
Addition of the product of the present invention in feedstuff is 60g/kg.
Using effect is tested
Using effect test using the probiotics of the embodiment of the present invention 1 in ablactational baby pig：
Experimental animal selects plant's average weight for the healthy pigletss 120 of (10.5 ± 0.45) kg, statistical analysis
Weight differences are not notable.Using single-factor Randomized Designs, by 120 healthy pigletss, by male and female half and half, be randomly divided into 2 groups it is (right
According to group and test group), per group of 6 repetitions, each repeats 10 pigs.Product of the present invention obtained in test group addition embodiment 1
45g/kg, matched group is without product of the present invention.Continuous feeding 20 days, compared with matched group, can using invention feed additive
Obtain following effect：
| Project | Test group | Matched group |
| Starting weight (kg) | 10.45 | 10.65 |
| End weight (kg) | 20.57 | 19.59 |
| Daily ingestion amount (g) | 720.1 | 705.2 |
| Daily gain (g) | 506 | 447 |
| Feedstuff-meat ratio | 1.42 | 1.58 |
| Diarrhea rate (head) | 6 | 24 |
Result of the test shows：After pigletss feedstuff of the feeding containing probiotics of the present invention, daily ingestion amount increases, and increases day by day and brings up again
Gao Liao, while also reducing the feedstuff-meat ratio of pigletss, diarrhea rate is significantly reduced.
Claims (7)
1. a kind of Bacillus coagulans, Lactobacillus plantarum and lactobacillus casei preparation, are prepared by following methods：
The preparation of seed liquor：By Bacillus coagulans (Bacillus coagulans) HM-09 30-38 DEG C in calorstat, 170
Turn shaking table culture 20-24h；By Lactobacillus plantarum (Lactobacillus plantarum) HM-10 and lactobacillus casei
(Lactobacillus casei) HM-09 30-38 DEG C of culture 20-24h in calorstat；
The seed culture medium of Bacillus coagulans HM-09, Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 is：Albumen
Peptone 2-5%, glucose 2-5%, adds water to 100%, pH7.0-7.2；
Fermentation culture conditions：Bacillus coagulans HM-09 seed liquor is added in solid medium to be carried out mixing after fermentation 0-
16h, inoculates Lactobacillus plantarum HM-10 seed liquor and continues fermentation 12-36h with lactobacillus casei HM-09 seed liquor, ferment altogether
Time 12-52h, controls fermentation temperature for 30-38 DEG C in sweat, humidity is 60-80%, fermentation ends its free ammonia ground state
Nitrogen content is in more than 480umol/g；
The solid medium is：25-30% bean cake, 25-30% wheat brans, 2-5% Semen Maydis powder, moisture 40-50%, initially
pH 6.8-7.2；
The Bacillus coagulans HM-09 deposit number is CGMCCNo.6729, and Lactobacillus plantarum HM-10 deposit numbers are
CGMCC No.6741, lactobacillus casei HM-09 deposit number is CGMCC No.6736.
2. Bacillus coagulans according to claim 1, Lactobacillus plantarum and lactobacillus casei preparation, it is characterised in that
3-6%, Lactobacillus plantarum HM-10 of the addition of the Bacillus coagulans HM-09 seed liquor for solid medium gross weight
2-4% of the addition of seed liquor for solid medium gross weight, the addition of lactobacillus casei HM-09 seed liquor are trained for solid
The 4-6% of foster base gross weight.
3. Bacillus coagulans according to claim 2, Lactobacillus plantarum and lactobacillus casei preparation, it is characterised in that
The addition of the Bacillus coagulans HM-09 seed liquor for solid medium gross weight 5%, Lactobacillus plantarum HM-10 kinds
The addition of sub- liquid for solid medium gross weight 3%, the addition of lactobacillus casei HM-09 seed liquor is solid medium
The 5% of gross weight.
4. the preparation method of a kind of Bacillus coagulans, Lactobacillus plantarum and lactobacillus casei preparation, comprises the steps：
The preparation of seed liquor：By Bacillus coagulans HM-09 30-38 DEG C in calorstat, 170 turns of shaking table cultures 20-24h；Will
Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 30-38 DEG C of culture 20-24h in calorstat；
The seed culture medium of Bacillus coagulans HM-09, Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 is：Albumen
Peptone 2-5%, glucose 2-5%, adds water to 100%, pH7.0-7.2；
Fermentation culture conditions：Bacillus coagulans HM-09 seed liquor is added in solid medium to be carried out mixing after fermentation 0-
16h, inoculates Lactobacillus plantarum HM-10 seed liquor and continues fermentation 12-36h with lactobacillus casei HM-09 seed liquor, ferment altogether
Time 12-56h, controls fermentation temperature for 30-38 DEG C in sweat, humidity is 60-80%, fermentation ends its free ammonia ground state
Nitrogen content is in more than 480umol/g；
The solid medium is：25-30% bean cake, 25-30% wheat brans, 2-5% Semen Maydis powder, moisture 40-50%, initially
pH 6.8-7.2；
The Bacillus coagulans HM-09 deposit number is CGMCC No.6729, and Lactobacillus plantarum HM-10 deposit numbers are
CGMCC No.6741, lactobacillus casei HM-09 deposit number is CGMCC No.6736.
5. the preparation method of Bacillus coagulans according to claim 4, Lactobacillus plantarum and lactobacillus casei preparation, its
It is characterised by, the 3-6% of the addition of the Bacillus coagulans HM-09 seed liquor for solid medium gross weight, plant breast
2-4% of the addition of bacillus HM-10 seed liquor for solid medium gross weight, the addition of lactobacillus casei HM-09 seed liquor
Measure the 4-6% for solid medium gross weight.
6. the preparation method of Bacillus coagulans according to claim 5, Lactobacillus plantarum and lactobacillus casei preparation, its
It is characterised by, the addition of the Bacillus coagulans HM-09 seed liquor is the 5% of solid medium gross weight, plant breast bar
The addition of bacterium HM-10 seed liquor for solid medium gross weight 3%, the addition of lactobacillus casei HM-09 seed liquor is
The 5% of solid medium gross weight.
7. the preparation method of Bacillus coagulans according to claim 4, Lactobacillus plantarum and lactobacillus casei preparation, its
It is characterised by, comprises the steps：
The preparation of seed liquor：Bacillus coagulans HM-09, Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 are from the sweet of cold storage
It is inoculated in oil pipe in the culture medium equipped with liquid seeds, the seed culture condition of Bacillus coagulans HM-09：37 DEG C,
170 turns of shaking table cultures 24h；Lactobacillus plantarum HM-10 and lactobacillus casei HM-09 37 DEG C of culture 24h in calorstat；
Wherein seed culture medium is：Peptone 2%, glucose 2%, adds water to 100%, pH7.0-7.2；
Solid fermentation
Fermentation culture conditions：Bacillus coagulans HM-09 seed liquor is added in solid medium carries out mixing after fermentation
12h, inoculates Lactobacillus plantarum HM-10 seed liquor and lactobacillus casei HM-09 seed liquor continues fermentation 24h, when fermenting altogether
Between 36h, fermentation temperature is controlled in sweat for 37 DEG C, humidity is 70%, and fermentation ends its free ammonical nitrogen contents is
489umol/g；
The solid medium is：29% bean cake, 29% wheat bran, 2% Semen Maydis powder, moisture 40%, initial pH7.0,115 DEG C
Autoclaving 25min；
The addition of the Bacillus coagulans HM-09 seed liquor for solid medium gross weight 5%, Lactobacillus plantarum HM-
The addition of 10 seed liquor for solid medium gross weight 3%, the addition of lactobacillus casei HM-09 seed liquor are trained for solid
The 5% of foster base gross weight.
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| CN109609410B (en) * | 2019-01-08 | 2021-07-30 | 天津市农业科学院 | Alfalfa silage starter and alfalfa silage preparation method |
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| CN115868581A (en) * | 2023-01-07 | 2023-03-31 | 山东省农业科学院畜牧兽医研究所 | Beef cattle fermented feed and preparation method thereof |
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