CN106434400A - Saccharomyces cerevisiae culture as well as preparation method and application of saccharomyces cerevisiae culture - Google Patents
Saccharomyces cerevisiae culture as well as preparation method and application of saccharomyces cerevisiae culture Download PDFInfo
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Abstract
The invention discloses a saccharomyces cerevisiae culture as well as a preparation method and application of saccharomyces cerevisiae culture. The preparation method comprises the following steps: performing seed culture of saccharomyces cerevisiae to obtain a saccharomyces cerevisiae seed culture; inoculating a liquid medium with the saccharomyces cerevisiae seed culture for aerobic fermentation to obtain a saccharomyces cerevisiae liquid culture; inoculating the saccharomyces cerevisiae liquid culture into a solid medium for anaerobic fermentation; after the fermentation, performing wall breaking treatment of the saccharomyces cerevisiae; and drying the saccharomyces cerevisiae solid culture after the wall breaking treatment to obtain the saccharomyces cerevisiae culture. Being as micro-ecological product replacing antibiotics, the saccharomyces cerevisiae culture disclosed by the invention is an urgent need of the breeding industry and plays an increasingly important role in ecological breeding production; and moreover, the development, production and application of the kind of products conform to the development situation of future animal husbandry of the country and have a broad market development prospect.
Description
Technical field
The present invention relates to field of feed, be specifically related to a kind of Cultures of S. cerevisiae and its preparation method and application.
Background technology
What antibiotic application was spread unchecked is global problem, the particularly relatively underdeveloped country of aquaculture, owing to raising
Bad environments, epidemic disease is often sent out, so problem is more serious.It is reported：China is annual produces antibiotic raw material about 210,000 tons, its
In 46.1% (9.7 ten thousand tons) be used in livestock breeding industry, be mainly used in animal rush increase and prophylactic effect, antibiosis
Public health, food security and environmental pollution are caused tremendous influence by the abuse of element, reduce antibiotic during livestock and poultry cultivation
Use have become as the problem demanding prompt solution that current livestock and poultry breeding industry kind faces.
Content of the invention
In view of this, it is an object of the invention to propose a kind of Cultures of S. cerevisiae and its preparation method and application, with
Minimizing or the use of substitute antibiotics, reduce antibiotic residue in animal products.
Based on the preparation method of the Cultures of S. cerevisiae that above-mentioned purpose, the present invention provide, comprise the following steps：
1) saccharomyces cerevisiae is carried out seed culture, obtain saccharomyces cerevisiae inoculum；
2) it is inoculated in saccharomyces cerevisiae inoculum in fluid nutrient medium and carries out aerobic fermentation, obtain saccharomyces cerevisiae liquid
Culture；
3) it is inoculated into saccharomyces cerevisiae liquid culture in solid medium and carries out anaerobic fermentation, to yeast after fermentation ends
Bacterium carries out broken wall treatment, then dries the saccharomyces cerevisiae solid culture after broken wall treatment, i.e. obtains described saccharomyces cerevisiae and cultivates
Thing.
In some embodiments of the invention, in described step 3) in, described solid medium consist of corn 5～
15%th, wheat bran 70～90%, dregs of beans 5～15%, by weight.
In some embodiments of the invention, in described step 3) in, by the saccharomyces cerevisiae liquid culture that obtains according to
The inoculum concentration of 10～20%v/m is inoculated in solid medium, anaerobic fermentation 12～48 hours under conditions of 25～40 DEG C.
In some embodiments of the invention, in described step 3) in, after anaerobic fermentation terminates, 55～65 DEG C of conditions
Under, keep temperature 12～24h, broken wall treatment is carried out to saccharomycete.
In some embodiments of the invention, in described step 3) in, use the method for pneumatic conveying drying to dry, dry
Leaving air temp is 60～80 DEG C, and charging rate is 1～4r/min.
In some embodiments of the invention, in described step 2) in, aerobic fermentation condition is：Fermentation temperature 25～40
DEG C, rotating speed 200～600rpm, throughput is 15～30L/min, and initial stage rotating speed is 200～300rpm, waits dissolved oxygen to drop to
Less than 15%, by adjusting rotating speed, make dissolved oxygen control more than 15～20%.
In some embodiments of the invention, in described step 2) in, in aerobic fermentation incubation, use stream to add benefit
Material mode fed-batch medium in described fluid nutrient medium, flow acceleration is 1000～1500ml/ hour, little in fermentation 8～12
Starting stream when after to add, stream adds 4～and 5 hours；
Wherein, fed-batch medium sugar concentration 20～28%, consists of urea 0.8～1.5%, and magnesium sulfate 0.05～
0.15%, calcium chloride 0.005～0.015%, potassium dihydrogen phosphate 0.05～0.15%, remaining is water, by weight, pH=5.5
～6.5.
The present invention also provides the Cultures of S. cerevisiae that the preparation method of a kind of above-mentioned Cultures of S. cerevisiae prepares.
The present invention also provides the application of a kind of above-mentioned Cultures of S. cerevisiae, and described Cultures of S. cerevisiae resists as an alternative
The Tiny ecosystem product of raw element makes an addition in daily ration.
In some embodiments of the invention, addition in daily ration for the described Cultures of S. cerevisiae is 0.5～5%.
From the above it can be seen that the Tiny ecosystem of the Cultures of S. cerevisiae antibiotic as an alternative of present invention offer produces
Product, are not only aquaculture reality and are badly in need of, play an increasingly important role in ecologic breeding produces, and the opening of such product
Send out, produce and application meets the following animal husbandry development present situation of China, there is wide market development prospect.
Detailed description of the invention
For making the object, technical solutions and advantages of the present invention clearer, below in conjunction with specific embodiment, to this
Bright further description.
Embodiment 1
As one embodiment of the present of invention, the preparation method of described Cultures of S. cerevisiae comprises the following steps：
(1) seed culture
Saccharomyces cerevisiae is transferred on YPD culture medium, wherein cultivate and consist of glucose the 2%th, yeast extract the 1%th, albumen
Peptone 2% (by weight), inoculum concentration is 10% (v/m), cultivates 16 hours in 30 DEG C of shaking table 200rpm.In the present embodiment, institute
Stating S. cervisiae is S. cervisiae (Saccharomyces cerevisiae) Sa-10, is preserved in China General Microbiological
Culture presevation administrative center, preserving number is CGMCC No.6120.
(2) liquid fermentation
The saccharomyces cerevisiae inoculum obtaining is inoculated in fluid nutrient medium with 8% inoculum concentration (v/m) and carries out aerobic
Fermentation, wherein, fluid nutrient medium sugar concentration is 1.5%, consists of urea 0.056%, magnesium sulfate 0.1%, calcium chloride 0.01%,
Potassium dihydrogen phosphate 0.1%, remaining is water (by weight), adjusts initial pH=5.5～5.8.Fermentation condition is：Constant temperature 30 DEG C,
Rotating speed 200～600rpm, initial stage rotating speed is 200rpm, waits dissolved oxygen to drop to less than 15%, by adjusting rotating speed, makes dissolved oxygen control
More than 15% (note：Demarcate when 100% demarcation rotating speed of dissolved oxygen electrode is 200rpm), throughput 20～25L/min.
During fermented and cultured, use flow feeding mode fed-batch medium in described fluid nutrient medium, wherein, stream
Add culture medium sugar concentration 24%, consist of urea 1.2%, magnesium sulfate 0.1%, calcium chloride 0.01%, potassium dihydrogen phosphate 0.1%,
Remaining is water (by weight), adjusts pH=5.5～6.0.Flow acceleration is 1200ml/ hour, starts after fermenting 10 hours
Stream adds, and stream adds 4 hours.
(3) solid fermentation
The saccharomyces cerevisiae liquid culture obtaining is inoculated on solid medium according to the inoculum concentration of 12% (v/m), presses
Weight meter, described solid medium consists of corn the 10%th, wheat bran the 80%th, dregs of beans 10%.Solid anaerobic under conditions of 33 DEG C
Ferment 36 hours, after anaerobic fermentation terminates, under the conditions of 58～62 DEG C, keep temperature 20h, sufficient broken wall is carried out to saccharomycete
Process.Then the saccharomyces cerevisiae solid culture after broken wall treatment is dried, i.e. obtain described Cultures of S. cerevisiae.Can adopt
Dried by the method for pneumatic conveying drying, dry leaving air temp 70 DEG C, charging rate 2.5r/min.
Embodiment 2
As one embodiment of the present of invention, the preparation method of described Cultures of S. cerevisiae comprises the following steps：
(1) seed culture
Saccharomyces cerevisiae is transferred on YPD culture medium, wherein cultivate and consist of glucose the 1.5%th, yeast extract the 1.2%th,
Peptone 2.1% (by weight), inoculum concentration is 8% (v/m), cultivates 17 hours in 32 DEG C of shaking table 250rpm.At the present embodiment
In, described S. cervisiae is S. cervisiae (Saccharomyces cerevisiae) Sa-10, is preserved in China commonly micro-
Biological inoculum preservation administrative center, preserving number is CGMCC No.6120.
(2) liquid fermentation
The saccharomyces cerevisiae inoculum obtaining is inoculated in fluid nutrient medium with 9% inoculum concentration (v/m) and carries out aerobic
Fermentation, wherein, fluid nutrient medium sugar concentration is 1.45%, consists of urea 0.052%, magnesium sulfate 0.08%, calcium chloride
0.012%, potassium dihydrogen phosphate 0.13%, remaining is water (by weight), adjusts initial pH=5.7～6.1.Fermentation condition is：
Constant temperature 32 DEG C, rotating speed 200～600rpm, initial stage rotating speed is 220rpm, waits dissolved oxygen to drop to less than 15%, by adjusting rotating speed,
Dissolved oxygen is made to control more than 17% (note：Demarcate when 100% demarcation rotating speed of dissolved oxygen electrode is 200rpm), throughput 18～
22L/min.
During fermented and cultured, use flow feeding mode fed-batch medium in described fluid nutrient medium, wherein, stream
Add culture medium sugar concentration 25%, consist of urea 1.05%, magnesium sulfate 0.08%, calcium chloride 0.011%, potassium dihydrogen phosphate
0.09%, remaining is water (by weight), adjusts pH=5.9～6.3.Flow acceleration was 1300ml/ hour, fermentation 9 hours
After start stream and add, stream adds 4.5 hours.
(3) solid fermentation
The saccharomyces cerevisiae liquid culture obtaining is inoculated on solid medium according to the inoculum concentration of 16% (v/m), presses
Weight meter, described solid medium consists of corn the 13%th, wheat bran the 76%th, dregs of beans 11%.Under conditions of 37.5 DEG C, solid is detested
Aerobe fermentation 40 hours, after anaerobic fermentation terminates, under the conditions of 60～65 DEG C, keeps temperature 13h, sufficiently breaks saccharomycete
Wall process.Then the saccharomyces cerevisiae solid culture after broken wall treatment is dried, i.e. obtain described Cultures of S. cerevisiae.Permissible
The method using pneumatic conveying drying is dried, and dries leaving air temp 73 DEG C, charging rate 2r/min.
Embodiment 3
As one embodiment of the present of invention, the preparation method of described Cultures of S. cerevisiae comprises the following steps：
(1) seed culture
Saccharomyces cerevisiae is transferred on YPD culture medium, wherein cultivate and consist of glucose the 2.3%th, yeast extract the 0.75%th,
Peptone 2% (by weight), inoculum concentration is 13% (v/m), cultivates 18 hours in 35 DEG C of shaking table 180rpm.At the present embodiment
In, described S. cervisiae is S. cervisiae (Saccharomyces cerevisiae) Sa-10, is preserved in China commonly micro-
Biological inoculum preservation administrative center, preserving number is CGMCC No.6120.
(2) liquid fermentation
The saccharomyces cerevisiae inoculum obtaining is inoculated in fluid nutrient medium with 5% inoculum concentration (v/m) and carries out aerobic
Fermentation, wherein, fluid nutrient medium sugar concentration is 1.7%, consists of urea 0.062%, magnesium sulfate 0.11%, calcium chloride
0.01%, potassium dihydrogen phosphate 0.07%, remaining is water (by weight), adjusts initial pH=6.3～6.5.Fermentation condition is：Permanent
Temperature 35 DEG C, rotating speed 200～600rpm, initial stage rotating speed is 250rpm, waits dissolved oxygen to drop to less than 15%, by adjusting rotating speed, makes
Dissolved oxygen controls more than 18% (note：Demarcate when 100% demarcation rotating speed of dissolved oxygen electrode is 200rpm), throughput 24～30L/
min.
During fermented and cultured, use flow feeding mode fed-batch medium in described fluid nutrient medium, wherein, stream
Add culture medium sugar concentration 22%, consist of urea 0.9%, magnesium sulfate 0.1%, calcium chloride 0.012%, potassium dihydrogen phosphate
0.09%, remaining is water (by weight), adjusts pH=5.8～6.2.Flow acceleration was 1500ml/ hour, fermentation 11 hours
After start stream and add, stream adds 4 hours.
(3) solid fermentation
The saccharomyces cerevisiae liquid culture obtaining is inoculated on solid medium according to the inoculum concentration of 18% (v/m), presses
Weight meter, described solid medium consists of corn the 15%th, wheat bran the 77%th, dregs of beans 8%.Under conditions of 35 DEG C, Solid anaerobic is sent out
Ferment 20 hours, after anaerobic fermentation terminates, under the conditions of 59～63 DEG C, keeps temperature 12h, carries out saccharomycete at sufficient broken wall
Reason.Then the saccharomyces cerevisiae solid culture after broken wall treatment is dried, i.e. obtain described Cultures of S. cerevisiae.Can use
The method of pneumatic conveying drying is dried, and dries leaving air temp 67 DEG C, charging rate 3.5r/min.
Embodiment 4
As one embodiment of the present of invention, the preparation method of described Cultures of S. cerevisiae comprises the following steps：
(1) seed culture
Saccharomyces cerevisiae is transferred on YPD culture medium, wherein cultivate and consist of glucose the 2%th, yeast extract the 1.25%th, egg
White peptone 1.85% (by weight), inoculum concentration is 12% (v/m), cultivates 17.5 hours in 28 DEG C of shaking table 200rpm.In this enforcement
In example, described S. cervisiae is S. cervisiae (Saccharomyces cerevisiae) Sa-10, is preserved in China common
Microbiological Culture Collection administrative center, preserving number is CGMCC No.6120.
(2) liquid fermentation
The saccharomyces cerevisiae inoculum obtaining is inoculated in fluid nutrient medium with 6% inoculum concentration (v/m) and carries out aerobic
Fermentation, wherein, fluid nutrient medium sugar concentration is 1.3%, consists of urea 0.052%, magnesium sulfate 0.08%, calcium chloride
0.01%, potassium dihydrogen phosphate 0.14%, remaining is water (by weight), adjusts initial pH=6.0～6.3.Fermentation condition is：Permanent
Temperature 28 DEG C, rotating speed 200～600rpm, initial stage rotating speed is 210rpm, waits dissolved oxygen to drop to less than 15%, by adjusting rotating speed, makes
Dissolved oxygen controls more than 19% (note：Demarcate when 100% demarcation rotating speed of dissolved oxygen electrode is 200rpm), throughput 18～22L/
min.
During fermented and cultured, use flow feeding mode fed-batch medium in described fluid nutrient medium, wherein, stream
Add culture medium sugar concentration 27.5%, consist of urea 1.5%, magnesium sulfate 0.06%, calcium chloride 0.013%, potassium dihydrogen phosphate
0.12%, remaining is water (by weight), adjusts pH=6.1～6.4.Flow acceleration was 1000ml/ hour, fermentation 12 hours
After start stream and add, stream adds 5 hours.
(3) solid fermentation
The saccharomyces cerevisiae liquid culture obtaining is inoculated on solid medium according to the inoculum concentration of 15% (v/m), presses
Weight meter, described solid medium consists of corn the 5%th, wheat bran the 80%th, dregs of beans 15%.Under conditions of 28 DEG C, Solid anaerobic is sent out
Ferment 45 hours, after anaerobic fermentation terminates, under the conditions of 58～62 DEG C, keeps temperature 19h, carries out saccharomycete at sufficient broken wall
Reason.Then the saccharomyces cerevisiae solid culture after broken wall treatment is dried, i.e. obtain described Cultures of S. cerevisiae.Can use
The method of pneumatic conveying drying is dried, and dries leaving air temp 78 DEG C, charging rate 3r/min.
Embodiment 5
As one embodiment of the present of invention, the preparation method of described Cultures of S. cerevisiae comprises the following steps：
(1) seed culture
Saccharomyces cerevisiae is transferred on YPD culture medium, wherein cultivate and consist of glucose the 1.5%th, yeast extract the 1.2%th,
Peptone 2% (by weight), inoculum concentration is 7% (v/m), cultivates 17.5 hours in 30 DEG C of shaking table 220rpm.At the present embodiment
In, described S. cervisiae is S. cervisiae (Saccharomyces cerevisiae) Sa-10, is preserved in China commonly micro-
Biological inoculum preservation administrative center, preserving number is CGMCC No.6120.
(2) liquid fermentation
The saccharomyces cerevisiae inoculum obtaining is inoculated in fluid nutrient medium with 5% inoculum concentration (v/m) and carries out aerobic
Fermentation, wherein, fluid nutrient medium sugar concentration is 1.38%, consists of urea 0.058%, magnesium sulfate 0.12%, calcium chloride
0.01%, potassium dihydrogen phosphate 0.11%, remaining is water (by weight), adjusts initial pH=5.8～6.2.Fermentation condition is：Permanent
Temperature 30 DEG C, rotating speed 200～600rpm, initial stage rotating speed is 200rpm, waits dissolved oxygen to drop to less than 15%, by adjusting rotating speed, makes
Dissolved oxygen controls more than 16% (note：Demarcate when 100% demarcation rotating speed of dissolved oxygen electrode is 200rpm), throughput 28～35L/
min.
During fermented and cultured, use flow feeding mode fed-batch medium in described fluid nutrient medium, wherein, stream
Add culture medium sugar concentration 22%, consist of urea 0.8%, magnesium sulfate 0.15%, calcium chloride 0.008%, potassium dihydrogen phosphate
0.05%, remaining is water (by weight), adjusts pH=5.9～6.4.Flow acceleration was 1450ml/ hour, fermentation 10 hours
After start stream and add, stream adds 4.5 hours.
(3) solid fermentation
The saccharomyces cerevisiae liquid culture obtaining is inoculated on solid medium according to the inoculum concentration of 14% (v/m), presses
Weight meter, described solid medium consists of corn the 12%th, wheat bran the 75%th, dregs of beans 13%.Solid anaerobic under conditions of 30 DEG C
Ferment 37 hours, after anaerobic fermentation terminates, under the conditions of 50～65 DEG C, keep temperature 15h, sufficient broken wall is carried out to saccharomycete
Process.Then the saccharomyces cerevisiae solid culture after broken wall treatment is dried, i.e. obtain described Cultures of S. cerevisiae.Can adopt
Dried by the method for pneumatic conveying drying, dry leaving air temp 64 DEG C, charging rate 1.5r/min.
The Cultures of S. cerevisiae obtaining according to embodiments of the present invention is had the aromatic odor of uniqueness, in subacidity.Make
Yeast cell wall in brewer yeast culture, after abundant broken wall treatment, makes the glucan in Cultures of S. cerevisiae and sweet dew gather
Sugar content improves, and the prebiotic component such as cellular content, end product of metabolism fully discharges, and these can make Cultures of S. cerevisiae play
Live products is not available or embodies insufficient food calling, improve the biological effects such as immunity, enteron aisle be prebiotic.Mannosan is ferment
The cell wall constituent of female bacterium, has immunocompetence, can activate the immunity improving animal.The molten albumen of acid is the relatively low egg of molecular weight
White hydrolysate, is the polypeptides matter that molecular weight is low, is more beneficial for animal and absorbs.
Comparative example 1：
(1) seed culture：With embodiment 1；
(2) liquid fermentation：With embodiment 1；
(3) solid fermentation
The saccharomyces cerevisiae liquid culture obtaining is inoculated on solid medium according to the inoculum concentration of 12% (v/m), presses
Weight meter, described solid medium consists of corn the 10%th, wheat bran the 80%th, dregs of beans 10%.After mixing, the condition of 33 DEG C
Lower solid anaerobic fermentation 36 hours, after anaerobic fermentation terminates, does not carry out broken wall treatment.Then by the saccharomyces cerevisiae after anaerobic fermentation
Solid culture is dried, and obtains Cultures of S. cerevisiae, the method for pneumatic conveying drying can be used directly to dry, and dries air-out
Temperature 70 C, charging rate 2.5r/min.
Comparative example 2：
(1) seed culture：With embodiment 1；
(2) liquid fermentation：With embodiment 1；
(3) solid medium mixes with liquid culture
The saccharomyces cerevisiae liquid culture obtaining is inoculated on solid medium according to the inoculum concentration of 12% (v/m), presses
Weight meter, described solid medium consists of corn 10%, wheat bran 80%, dregs of beans 10%.After mixing, do not carry out solid and send out
Mixture is directly dried, is obtained Cultures of S. cerevisiae by ferment and broken wall treatment.The method that can use pneumatic conveying drying is directly entered
Row is dried, and dries leaving air temp 70 DEG C, charging rate 2.5r/min.
Embodiment 1-5, after obtaining the high liquid spawn of a large amount of activity, uses liquid aerobic fermentation and solid anaerobic fermentation
And the technique that Solid anaerobic broken wall treatment combines, comparative example 1, after obtaining the high liquid spawn of a large amount of activity, uses liquid to have
Aerobe fermentation and solid anaerobic fermentation technique；Comparative example 2, after obtaining the high liquid spawn of a large amount of activity, uses liquid aerobic fermentation
Technique, after solid medium mixes with liquid culture, directly carries out drying and processing without fermentation.By embodiment 1-5
The yeast culture obtaining with comparative example 1-2 carries out nutritive index analysis, and analysis result is shown in Table 1.
The nutritive index of table 1 each embodiment Cultures of S. cerevisiae
Note：The English alphabet of numerical value upper right corner mark, colleague is different, and lowercase alphabet shows significant difference (P<0.05).
Utilize SPSS DAS to carry out significance difference specific analysis, from table 1, send out with single liquid aerobic
Ferment, solid anaerobic fermentation are compared without broken wall treatment, and the Cultures of S. cerevisiae being obtained by the preparation method of the present invention is carried
The high nutritional labeling of yeast culture, especially significantly improves mannosan and acid molten protein content water in yeast culture
Flat.
During modern livestock and poultry cultivation produces, people, by adding medicated feed additive (i.e. antibiotic) in feed, come pre-
Livestock and poultry are played the effect promoting growth by bacteriosis that is anti-or that treat livestock and poultry simultaneously.Conventional resisting in piglet production process
Life have Nosiheptide, colistine sulfate, olaquindox etc..The present invention provides the application that a kind of above-mentioned saccharomyces cerevisiae is cultivated, described
The Tiny ecosystem product of Cultures of S. cerevisiae antibiotic as an alternative makes an addition in daily ration.Alternatively, described saccharomyces cerevisiae is cultivated
Addition in daily ration for the thing is 0.5～5%.It is preferred that the addition that described Cultures of S. cerevisiae is in daily ration is
0.8%.Addition in daily ration for the described Cultures of S. cerevisiae is 1%.Described Cultures of S. cerevisiae adding in daily ration
Dosage is 2%.Addition in daily ration for the described Cultures of S. cerevisiae is 2.5%.
The Cultures of S. cerevisiae that the present invention provides is with saccharomyces cerevisiae as bacterial classification, after solid fermentation, concentrates, is dried and obtains
The product obtaining.Cultures of S. cerevisiae as a kind of feedstuff, can for animal provide abundant fermentation substrate, mycoprotein,
The prebiotic substances such as metabolism of yeasts product, yeast cell wall, have good to the intestinal growth of animal, the colony balance of enteron aisle
Facilitation.The present invention, by adding Cultures of S. cerevisiae in pig starter feed, reduces the quantity of antibiotic in feed simultaneously
Or kind.
1 materials and methods
1.1 test raw material
Feed medicated premix:Nosiheptide premixed agent：Specification 1%, is produced by Zhejiang Province Huineng animal drugs Co., Ltd；Sulphur
Acid colistin pre-mixing agent：Specification 10%, is produced by bio tech ltd incomparably, Zhejiang；Olaquindox pre-mixing agent：Specification
50%, produced by Huang gang animal drug factory of Zhongmu Industry Co., Ltd.
Cultures of S. cerevisiae：The Cultures of S. cerevisiae being prepared by above-described embodiment.
10% composite pre-mixed fodder for piglets：Being there is provided by Beijing YingHuiEr Bioisystech Co., Ltd, every 1 kilogram of premix contains
Have：Vitamin A 96000IU, vitamin D312000IU, vitamin e1 80mg, vitamin K38.6mg, Cobastab117mg, dimension are raw
Element B240mg, Cobastab621mg, Cobastab120.2mg, nicotinic acid 196mg, calcium pantothenate 129mg, folic acid 5.9mg, biotin
0.6mg, Choline Chloride 4400mg, lysine the 2.5%th, methionine the 1.0%th, iron 1920mg, copper 1700mg, zinc 1100mg, manganese
250mg, iodine 2mg, selenium 2.5mg, total phosphorus the 1.0%th, phytase 7500U.
1.2 experimental design and packet
Select 180 42 age in days child care piglets, be randomly divided into 4 groups, respectively control group, test 1 group, test 2 groups, examination
Testing 3 groups and test 4 groups, often organizing 5 repetitions, each repeats 9.
Experimental design is shown in Table 1.
Table 2 experimental design unit：Mg/kg daily ration
| Feed medicated premix | Control group | Test 1 group | Test 2 groups | Test 3 groups | Test 4 groups |
| Nosiheptide | 5 | 5 | 5 | 5 | 0 |
| Colistine sulfate | 20 | 10 | 0 | 0 | 0 |
| Olaquindox | 100 | 50 | 100 | 0 | 0 |
Note：Feed medicated premix content is all as active ingredients.
1.3 test daily rations
Test Diet Formula is shown in Table 2.
Diet Formula unit tested by table 3：%
| Raw material | Control group | Test 1 group | Test 2 groups | Test 3 groups | Test 4 groups |
| Corn | 64 | 63 | 63 | 63 | 63 |
| Dregs of beans (crude protein 43%) | 18 | 18 | 18 | 18 | 18 |
| Expanded soybean | 8 | 8 | 8 | 8 | 8 |
| Cultures of S. cerevisiae | 0 | 1 | 1 | 1 | 1 |
| 10% composite pre-mixed fodder for piglets | 10 | 10 | 10 | 10 | 10 |
| Add up to | 100 | 100 | 100 | 100 | 100 |
Note：All of feed medicated premix all makes an addition in 10% composite pre-mixed fodder for piglets.
1.4 trial
Test on April 18th, 2016 to May 17 Beijing nine ancient cooking vessel breeding cultivation Co., Ltd carry out.
Carry out to each group of piglet morning April 18 weighing and being grouped on an empty stomach, during test, record each feed consumption rate repeating,
Record each diarrhoea head number repeating and dead head number.Evening 23 May 17：00 stops material, and the clout amount in cleaning hopper simultaneously claims
Weight, carries out to piglet weighing on an empty stomach morning May 18, off-test.
Piglet free choice feeding and drinking-water during test, immune programme for children is carried out according to pig farm operational procedure.
1.5 testing index
1.5.1 test daily ration testing index
Often organize daily ration and take 200g, measure crude protein, coarse ash, calcium, phosphorus, sodium chloride and crude fat.
1.5.2 piglet growth performance testing index
The average daily ingestion amount of piglet, average daily gain, feedstuff-meat ratio, diarrhea rate and the death rate.
1.6 statistical analysis
SPSS is used to carry out one-way analysis of variance (One-way ANOVA) and Duncan ' s multiple ratio to test data
Relatively, result is represented by " mean+SD ", and the significance of equal value difference is p<0.05.
2 results and analysis
2.1 test daily rations measure
Daily ration measurement result unit tested by table 4：%
| Index | Control group | Test 1 group | Test 2 groups | Test 3 groups | Test 4 groups |
| Crude protein | 17.57 | 17.86 | 17.31 | 16.99 | 17.24 |
| Coarse ash | 4.66 | 4.77 | 4.68 | 4.98 | 4.53 |
| Calcium | 0.94 | 0.86 | 0.77 | 0.80 | 0.82 |
| Total phosphorus | 0.55 | 0.55 | 0.54 | 0.57 | 0.56 |
| Sodium chloride | 0.45 | 0.41 | 0.41 | 0.40 | 0.40 |
| Crude fat | 5.35 | 5.40 | 5.40 | 5.53 | 5.38 |
As shown in Table 3, the nutrient component determining value of each group daily ration is close and substantially conforms to design load requirement.
2.2 piglet growth performance
Table 5 piglet growth performance measurement result
| Index | Control group | Test 1 group | Test 2 groups | Test 3 groups | Test 4 groups |
| First starting weight, kg | 13.34±1.42a | 13.35±1.23a | 13.36±1.23a | 13.36±1.13a | 13.35±1.20a |
| Terminate weight, kg | 29.07±2.27a | 29.16±2.02a | 28.77±2.01a | 30.26±2.04a | 31.44±2.07a |
| Average daily gain, g | 524±35a | 527±43a | 514±47a | 563±32a | 603±38a |
| Average daily ingestion amount, g | 937±68a | 942±84a | 943±82a | 985±62a | 1050±56a |
| Feedstuff-meat ratio | 1.79±0.03a | 1.79±0.11a | 1.84±0.07a | 1.75±0.07a | 1.74±0.12a |
| Diarrhea rate, % | 3.63±1.78a | 6.74±2.45a | 4.81±2.21a | 4.67±2.56a | 4.45±2.08a |
Note：Same row shoulder head letter identical table differential different not notable (P ＞ 0.05), letter is different on the shoulders represents difference
Significantly (p ＜ 0.05).
As shown in Table 5, that there are no significant is poor for the average daily gain between each group, average daily ingestion amount, feedstuff-meat ratio, diarrhea rate
Different.
Testing 1 group of colistine sulfate and olaquindox all halving than control group, the saccharomyces cerevisiae simultaneously adding 1% is cultivated
Thing, average daily gain, average daily ingestion amount are higher 3g and 5g than control group respectively；And the diarrhea rate testing 1 group increases than control group
About 3.1 percentage points.
Test 2 groups of olaquindox additions identical with control group, but without colistine sulfate, and add the wine brewing of 1%
Yeast culture, average daily gain low 10g than control group, low by 1.9%, average daily ingestion amount high 6g, Gao Liao than control group
0.6%；Feedstuff-meat ratio is higher by 0.05 than control group, improves 2.8%；Diarrhea rate adds about 1.2 percentage points than control group.
Test 3 groups of colistine sulfates and olaquindox all without only reservation Nosiheptide, simultaneously the wine brewing ferment of interpolation 1%
Female culture, average daily gain and average daily ingestion amount are higher 39g and 48g than control group respectively, have been respectively increased 7.4% He
5.1%；Feedstuff-meat ratio reduces by 0.04 than control group, reduces 2.2%；Diarrhea rate adds about 1.0 percentage points than control group.
Test 4 groups without any antibiotic, Cultures of S. cerevisiae, average daily gain and the average day of an interpolation 1%
Feed intake is higher 79g and 113g than control group respectively, has been respectively increased 15% and 12%；Feedstuff-meat ratio reduces 0.05, reduces
2.8%；Diarrhea rate adds 0.82 percentage point than control group.
2.3 discuss
Disease that bacterium, virus can be caused by the beta glucan containing in yeast cell wall and manna oligosacchride and environment because of
The stress reaction that element causes produces nospecific immunity, thus improves the immunity of piglet, the intestinal health of protection piglet, thus
Play disease-resistant, the somatotrophic effect identical with antibiotic.Test 1 group and test interpolation Nosiheptide and wine brewing ferment in 2 groups of daily rations
Female culture, and test 1 group of colistine sulfate and the addition of olaquindox halves simultaneously, test 2 groups and glue bar without sulfuric acid
Rhzomorph, two groups of piglets all can reach the growth performance close with control group, and diarrhea rate does not has significant difference yet.
Saccharomycete can produce the metabolite such as several amino acids, nucleotides during the fermentation, and beneficially alimentary canal is beneficial
The propagation of bacterium, promotes digesting and assimilating of enteral nutritional support material, can increase the feed intake of piglet, improves the profit to feed for the piglet
By rate, improve the production performance of piglet.Test and 3 groups of daily rations only add Nosiheptide and Cultures of S. cerevisiae, and without sulphur
Acid colistin and olaquindox, test in 4 groups of daily rations without any antibiotic, only adds Cultures of S. cerevisiae, and two groups young
The growth performance of pig has a higher daily ingestion amount and daily gain more simultaneously than the control group adding three kinds of antibiotic on the contrary, thus table
Reveal more excellent growth performance.Adding 5g/kg yeast culture in daily ration may be by improving height of naps, intestine immunity
Response and nutrient digestibility improve the growth performance of pig, also indicate that yeast culture can substitute in weanling pig daily ration simultaneously
AGP (duomycin).
The diarrhoea to piglet for the antibiotic has certain prevention effect, and 4 test group of the present invention are by adding saccharomyces cerevisiae training
When foster thing reduces addition or the kind of antibiotic, the diarrhea rate of piglet has certain increase, but remains at normal level,
Visible Cultures of S. cerevisiae also can effectively substitute antibiotics and the effect of playing prevention of diarrhea in piglets.Yeast culture can ensure
Piglet by stress after avoid suffering from diarrhoea, or speedy recovery to health level after suffering from diarrhoea.
3 conclusions
Piglet cradling phase daily ration only adds Cultures of S. cerevisiae, by equal for the addition of colistine sulfate and olaquindox
Halve or without colistine sulfate, olaquindox and Nosiheptide, do not affect the growth performance of piglet.Wherein only add that
The piglet of western peptide and Cultures of S. cerevisiae and the piglet performance only adding Cultures of S. cerevisiae without any antibiotic
Go out excellent growth performance, show the consumption using Cultures of S. cerevisiae can reduce antibiotic, it might even be possible to substitute antibiosis
The use of element.
The Cultures of S. cerevisiae that the present invention provides is that a kind of pollution-free, noresidue, rush are digested and assimilated, and growth promotion improves
The natural single feed raw material of immunity.It is by screening high-performance saccharomyces cerevisiae bacterial classification, under specific process conditions, adopts
Use specific solid medium, after anaerobic fermentation sufficient under specified conditions, concentrated, dry product again.It is permissible
It is nutritional substrate by being added with beneficial microorganism for animal intestinal, regulates microecological balance, promote digestion, improve the immunity of enteron aisle
Function, the use of minimizing or substitute antibiotics, reduces antibiotic residue in animal products, improves efficiency of feed utilization, save the energy
Reduce and pollute, ensure the optimal performance of animal health and the level of production.
As can be seen here, the Tiny ecosystem product of the Cultures of S. cerevisiae antibiotic as an alternative that the present invention provides, is not only
Aquaculture reality be badly in need of, ecologic breeding produce in play an increasingly important role, and the exploitation of such product, production and
Application meets the following animal husbandry development present situation of China, has wide market development prospect.
Those of ordinary skill in the field should be understood：The discussion of any of the above embodiment is exemplary only, not
It is intended to imply that the scope of the present disclosure (including claim) is limited to these examples；Under the thinking of the present invention, above example
Or also can be combined between the technical characteristic in different embodiments, step can realize with random order, and exists such as
Other changes of the many of the different aspect of the upper described present invention, in order to concisely they do not provide in details.Therefore, all
Within the spirit and principles in the present invention, any omission of being made, modification, equivalent, improvement etc., should be included in the present invention's
Within protection domain.
Claims (10)
1. the preparation method of a Cultures of S. cerevisiae, it is characterised in that comprise the following steps：
1) saccharomyces cerevisiae is carried out seed culture, obtain saccharomyces cerevisiae inoculum；
2) it is inoculated in saccharomyces cerevisiae inoculum in fluid nutrient medium and carries out aerobic fermentation, obtain saccharomyces cerevisiae Liquid Culture
Thing；
3) it is inoculated into saccharomyces cerevisiae liquid culture in solid medium and carries out anaerobic fermentation, after fermentation ends, saccharomycete is entered
Row broken wall treatment, then the saccharomyces cerevisiae solid culture after broken wall treatment is dried, i.e. obtain described Cultures of S. cerevisiae.
2. the preparation method of Cultures of S. cerevisiae according to claim 1, it is characterised in that in described step 3) in,
Described solid medium consist of corn 5～15%, wheat bran 70～90%, dregs of beans 5～15%, by weight.
3. the preparation method of Cultures of S. cerevisiae according to claim 1, it is characterised in that in described step 3) in,
The saccharomyces cerevisiae liquid culture obtaining is inoculated in solid medium according to the inoculum concentration of 10～20%v/m, 25～40
DEG C under conditions of anaerobic fermentation 12～48 hours.
4. the preparation method of Cultures of S. cerevisiae according to claim 1, it is characterised in that in described step 3) in,
After anaerobic fermentation terminates, under the conditions of 55～65 DEG C, keep temperature 12～24h, broken wall treatment is carried out to saccharomycete.
5. the preparation method of Cultures of S. cerevisiae according to claim 1, it is characterised in that in described step 3) in,
The method using pneumatic conveying drying is dried, and drying leaving air temp is 60～80 DEG C, and charging rate is 1～4r/min.
6. the preparation method of Cultures of S. cerevisiae according to claim 1, it is characterised in that in described step 2) in,
Aerobic fermentation condition is：Fermentation temperature 25～40 DEG C, rotating speed 200～600rpm, throughput is 15～30L/min, and the initial stage turns
Speed is 200～300rpm, waits dissolved oxygen to drop to less than 15%, by adjusting rotating speed, makes dissolved oxygen control more than 15～20%.
7. the preparation method of Cultures of S. cerevisiae according to claim 1, it is characterised in that in described step 2) in,
In aerobic fermentation incubation, using flow feeding mode fed-batch medium in described fluid nutrient medium, flow acceleration is
1000～1500ml/ hour, starts stream in fermentation and adds after 8～12 hours, and stream adds 4～and 5 hours；
Wherein, fed-batch medium sugar concentration 20～28%, consists of urea 0.8～1.5%, magnesium sulfate 0.05～0.15%, chlorine
Changing calcium 0.005～0.015%, potassium dihydrogen phosphate 0.05～0.15%, remaining is water, by weight, pH=5.5～6.5.
8. the preparation method of the Cultures of S. cerevisiae according to any one in claim 1～7 prepares
Cultures of S. cerevisiae.
9. the application of a Cultures of S. cerevisiae according to claim 8, it is characterised in that described saccharomyces cerevisiae is cultivated
The Tiny ecosystem product of thing antibiotic as an alternative makes an addition in daily ration.
10. the application of Cultures of S. cerevisiae according to claim 9, it is characterised in that described Cultures of S. cerevisiae
Addition in daily ration is 0.5～5%.
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Cited By (5)
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| CN107937292A (en) * | 2017-11-24 | 2018-04-20 | 北京英惠尔生物技术有限公司 | A kind of Cultures of S. cerevisiae and its zymotechnique |
| CN108887515A (en) * | 2018-09-06 | 2018-11-27 | 北京中农优嘉生物科技有限公司 | A kind of layer chicken feed |
| CN108936002A (en) * | 2018-01-09 | 2018-12-07 | 北京英惠尔生物技术有限公司 | Ferment liquid sow material and preparation method thereof |
| CN110150457A (en) * | 2018-09-25 | 2019-08-23 | 北京中农弘科生物技术有限公司 | A kind of antibacterial type yeast culture and its application |
| CN114181844A (en) * | 2021-09-02 | 2022-03-15 | 中粮面业（濮阳）有限公司 | Saccharomyces cerevisiae and leaven for improving flavor of noodles, application of saccharomyces cerevisiae and leaven and method for preparing flour products through fermentation |
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| CN107937292A (en) * | 2017-11-24 | 2018-04-20 | 北京英惠尔生物技术有限公司 | A kind of Cultures of S. cerevisiae and its zymotechnique |
| CN108936002A (en) * | 2018-01-09 | 2018-12-07 | 北京英惠尔生物技术有限公司 | Ferment liquid sow material and preparation method thereof |
| CN108887515A (en) * | 2018-09-06 | 2018-11-27 | 北京中农优嘉生物科技有限公司 | A kind of layer chicken feed |
| CN110150457A (en) * | 2018-09-25 | 2019-08-23 | 北京中农弘科生物技术有限公司 | A kind of antibacterial type yeast culture and its application |
| CN110150457B (en) * | 2018-09-25 | 2022-05-17 | 北京中农弘科生物技术有限公司 | Bacteriostatic yeast culture and application thereof |
| CN114181844A (en) * | 2021-09-02 | 2022-03-15 | 中粮面业（濮阳）有限公司 | Saccharomyces cerevisiae and leaven for improving flavor of noodles, application of saccharomyces cerevisiae and leaven and method for preparing flour products through fermentation |
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