CN111961602A - Saccharomyces cerevisiae and application thereof in feed for lactating calves - Google Patents
Saccharomyces cerevisiae and application thereof in feed for lactating calves Download PDFInfo
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Abstract
The invention is suitable for the technical field of microbial breeding, and provides a strain of Saccharomyces cerevisiae, wherein the Saccharomyces cerevisiae is Saccharomyces cerevisiae PLBS011(Saccharomyces cerevisiae PLBS011) which is preserved in the common microorganism center of China general microbiological culture Collection management Committee for preservation in 2018, 12 and 17 months, and the preservation number is CGMCC NO. 169951; the invention also provides application of the saccharomyces cerevisiae in feed for lactating calves. Therefore, the saccharomyces cerevisiae PLBS011 disclosed by the invention can improve the structure of the organism flora of the lactating calf, improve the immunity and production performance of the organism, and is beneficial to increasing the economic benefit of breeding the lactating calf.
Description
Technical Field
The invention relates to the technical field of microbial breeding, in particular to saccharomyces cerevisiae and application thereof in feed for lactating calves.
Background
The young stage is an important physiological period of the ruminant, and the growth and development of the stage are closely related to the potential production performance of the ruminant. As the gastrointestinal microorganisms and the immune system of the newborn calf are not healthy enough, the newborn calf is easy to have diarrhea in lactation period, which accounts for about 80% of the morbidity of the calf, and the newborn calf with diarrhea is easy to be infected with respiratory tract and intestinal tract diseases in the growth and development process in future, so that the development, growth, survival and the like of the calf are greatly influenced, the breeding and production performance of the calf in the later period can be directly influenced, and the economic benefit of the industry is further influenced.
Researches show that the yeast culture is rich in flavor-enhancing substances such as polypeptide, glucose, vitamins, nucleotide, alcohol lipid and the like, and can improve the feed intake and the production performance of animals; meanwhile, the micro-ecological balance in the intestinal tract can be adjusted or maintained, the immune function is enhanced, and the digestion and absorption of nutrient substances are promoted, so that the application effects of disease resistance and growth promotion are achieved. The saccharomyces cerevisiae is one of saccharomycetes, is rich in functional molecules such as organic acid, amino acid and the like capable of regulating an animal intestinal microflora, and can achieve the effects of improving the environment in the gastrointestinal tract and promoting digestion and absorption of nutrient components of feed by reducing the pH of an intestinal tract, inhibiting the growth of harmful bacteria, promoting the proliferation of beneficial bacteria such as lactic acid bacteria, cellulolytic bacteria and the like.
However, the application research of saccharomyces cerevisiae in the feed for the lactating calves is only reported, and the addition amount and the effect are different.
In view of the above, the prior art is obviously inconvenient and disadvantageous in practical use, and needs to be improved.
Disclosure of Invention
Aiming at the defects, the invention aims to provide the saccharomyces cerevisiae and the application thereof in the feed of the lactating calves, the saccharomyces cerevisiae PLBS011 has better fermentation performance and has good ethanol tolerance, acid resistance and temperature resistance, and the saccharomyces cerevisiae PLBS011 with a proper level is added in the daily ration of the lactating calves, so that the structure of the flora of the lactating calves can be improved, the immunity of the calves can be improved, the morbidity of the calves can be reduced, meanwhile, the utilization rate of the feed and the absorption rate of nutrient substances of the lactating calves can be improved, the production performance of the lactating calves can be improved, and the economic benefit of the breeding of the lactating calves can be increased.
In order to achieve the aim, the invention provides a strain of Saccharomyces cerevisiae, wherein the Saccharomyces cerevisiae PLBS011(Saccharomyces cerevisiae PLBS011) is preserved in China general microbiological culture collection center (CGMCC NO. 16951) in 2018, 12 and 17 months.
According to the saccharomyces cerevisiae of the present invention, the saccharomyces cerevisiae is obtained from a process of brewing rice wine from rice.
According to the saccharomyces cerevisiae, the maximum ethanol inhibition concentration of the saccharomyces cerevisiae is 9% vol.
According to the saccharomyces cerevisiae, the maximum inhibition temperature of the saccharomyces cerevisiae is 42 ℃.
According to the saccharomyces cerevisiae, the addition amount of the saccharomyces cerevisiae in the basic ration of the lactating calf is 1-5%.
According to the saccharomyces cerevisiae, the addition amount of the saccharomyces cerevisiae in the basic ration of the lactating calf is 2.3-3.5%.
According to the saccharomyces cerevisiae, the invention also provides a method for screening the saccharomyces cerevisiae, which comprises the following steps:
step one preparation of a diluent
Saccharifying rice for 72h, sealing and fermenting for 40 days at 25-35 ℃, and diluting fermentation liquor to obtain a diluent;
step two isolation of the strains
Coating the diluent in a Bengal culture medium, culturing for 2-4 days at 25-30 ℃, selecting a single colony with typical yeast colony characteristics, and separating and purifying to obtain an isolated strain;
step three primary screen
Taking the multiple strains obtained in the second step in sequence, diluting the strains, inoculating the diluted strains on a lower layer culture medium plate of a TTC method, and covering an upper layer culture medium on the lower layer culture medium when the bacterial colony grows to 1-2 mm; continuously culturing for 2-4 h, taking out, and selecting red strains to be respectively transferred to test tube slopes according to the color development condition of bacterial colonies;
step four double sifting
And (4) inoculating the red bacterial strain selected in the step three into an YPD liquid culture medium, and continuously performing activation culture for 20-30 h at the temperature of 25-35 ℃ to obtain the re-screened bacterial strain.
According to the method for screening the saccharomyces cerevisiae, in the second step, the separation and purification times of the strain are 2-3.
According to the saccharomyces cerevisiae disclosed by the invention, the saccharomyces cerevisiae is applied to feed for lactating calves.
The saccharomyces cerevisiae PLBS011 provided by the invention has better fermentation performance, good ethanol tolerance, acid resistance and temperature resistance, and the saccharomyces cerevisiae PLBS011 with a proper level is added into the daily ration of the lactating calf, so that the structure of the organism flora of the lactating calf can be improved, the immunity of the organism can be improved, the morbidity of the organism can be reduced, meanwhile, the utilization rate of the lactating calf to the feed and the absorption rate of nutrient substances can be improved, the production performance of the lactating calf can be improved, and the economic benefit of the breeding of the lactating calf can be increased.
Drawings
FIG. 1 is a colony morphology of Saccharomyces cerevisiae PLBS011 of the present invention;
FIG. 2 is a graph showing the growth of Saccharomyces cerevisiae PLBS011 according to the present invention;
FIG. 3 shows the absorbance values of the fermentation broth of Saccharomyces cerevisiae PLBS011 at a wavelength of 600 nm;
FIG. 4 is a graph of ethanol tolerance of Saccharomyces cerevisiae PLBS011 according to the present invention;
FIG. 5 is a graph of the acid resistance of Saccharomyces cerevisiae PLBS011 according to the present invention;
FIG. 6 is a temperature capability graph of Saccharomyces cerevisiae PLBS011 according to the present invention;
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to the accompanying drawings. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The invention provides a strain of saccharomyces cerevisiae, the scientific name of Latin: saccharomyces Cerevisiae; the Saccharomyces cerevisiae is Saccharomyces cerevisiae PLBS011(Saccharomyces cerevisiae PLBS011), which is preserved in China general microbiological culture Collection center (CGMCC for short, with the address of No. 3 Siro 1 of Beijing, Chaoyang district, North Cheng) in 12 and 17 days of 2018, and the preservation number is CGMCC NO. 16911.
The invention provides a method for screening saccharomyces cerevisiae PLBS011, which is a process of brewing rice wine by using saccharomyces cerevisiae taken from rice, and comprises the following steps:
step one preparation of a diluent
Saccharifying rice for 72h, sealing and fermenting for 40 days at 25-35 ℃, and diluting the fermentation liquor with sterile water to obtain a diluted solution.
Step two isolation of the strains
And coating the diluent in a Bengal red culture medium, culturing for 2-4 days at 25-30 ℃, then randomly selecting a single colony with typical yeast colony characteristics, and separating and purifying for 2-3 times to obtain the separated strain.
Step three primary screen
Sequentially collecting the obtained strains, and diluting by plate dilution method to obtain concentration gradient of 10-5Inoculating the strain on a lower layer culture medium plate of a TTC method, culturing for 45-60 h at 25-35 ℃, and covering a certain amount of upper layer culture medium on the lower layer culture medium when bacterial colony grows to 1-2 mm. Culturing for 2-4 h at 25-35 ℃ under a dark condition, taking out, immediately observing the coloration condition, and selecting red strains to be respectively transferred to the inclined planes of the test tubes according to the coloration condition of colonies.
TTC is (2,3, 5-triphenyltetrazolium chloride) color-developing agent, can generate color reaction on metabolites of yeast, and can judge the enzyme activity of the yeast, namely the alcohol production capacity of yeast fermentation. A layer of TTC color developing agent is covered on yeast colonies cultured on a certain culture medium, TTC can show different colors, and yeast with strong alcohol production capacity can show deep red, next pink, reddish or non-colored wild yeast.
Step four double sifting
And (3) inoculating the primarily screened red strain into an YPD liquid culture medium, and continuously performing activation culture for 20-30 h at the temperature of 25-35 ℃ to obtain a re-screened strain.
After the re-screened strains are screened by the method, the strains are identified, and the invention provides an identification method of yeast strains, which comprises the following steps:
morphological identification
According to the standard method for identifying yeasts in the fourth edition of Classification of yeasts, the size and morphology of cells, the texture of colonies, the color of colonies, the surface characteristics of colonies, the edges of colonies, the reproductive characteristics and the like are observed by using an optical microscope and a stereoscope, and morphological identification is performed.
The form of the re-screened strain is as follows: the surface is smooth, moist and sticky, the texture is uniform, the colors of the front and back sides, the edges and the central part are uniform, and the bacterial colony is milky white. (see FIG. 1)
Identification of Gene sequences
Performing molecular biological identification on the re-screened strain, extracting the genome DNA of the re-screened strain, and designing a universal primer according to the conservation of the 26S rDNA D1/D2 region sequence of the fungus: the front primer 5'-CAGAGTTTGATCCTGGCT-3' and the rear primer 5'-AGGAGGTGATCCAGCCGCA-3' are used for PCR amplification, and PCR products are sent to the company of Biotechnology engineering (Shanghai) Ltd for sequencing. The 5 'end and the 3' end are two ends of the gene, and after the DNA template is opened, the primer is combined with the template and is copied from the 5 'end to the 3' end.
Through gene sequence identification, the re-screened strain is classified as saccharomycete and named as saccharomyces cerevisiae PLBS 011.
In order to verify the strain properties of the Saccharomyces cerevisiae PLBS011 of the present invention, strain property tests including growth curve, fermentation performance and tolerance performance tests of the strain were performed.
Growth curve of the Strain
Inoculating the strain into a sterilized YPD liquid culture medium, carrying out shake culture at a constant temperature of 25-35 ℃, observing the production condition of the strain for 72h, sampling at an interval of 2h at the early stage, sampling at intervals of 4, 6 and 12h at the later stage, measuring the absorbance of the strain at a wavelength of 560nm by using a visible spectrophotometer, repeating for 5 times, and drawing the change trend of a strain growth curve (see figure 2).
According to the growth curve, the bacterial quantity is exponentially increased between 2-20 h of culture, the bacterial quantity reaches a stable state after 20h, and the growth of the microzyme is stable in a longer time (72 h).
Fermentation Performance of the Strain
Inoculating the strain activated to logarithmic phase into a fermentation medium with a substrate mass concentration of 75-85 g/L according to an inoculation amount of 5% and a liquid loading amount of 100mL/250mL, culturing for 72h, sampling every 12h, and determining the absorbance value of the fermentation liquid under the condition of 600nm wavelength (see figure 3).
From the absorbance value curve, OD600nmThe value increased rapidly to 6.7 in the first 24h and then slowly. OD600nmThe overall change of the value is that the value is increased continuously along with the extension of the fermentation time, reaches the maximum at 24h and keeps approximately stable, which indicates that the fermentation performance of the saccharomyces cerevisiae PLBS011 is better.
Yeast is a commonly used fermentation strain in livestock feed, but the tolerance of yeast often becomes an important factor for limiting the fermentation effect of the yeast on the feed, so the invention tests the tolerance of the saccharomyces cerevisiae PLBS011, mainly comprising ethanol tolerance, acid resistance and temperature resistance.
Determination of ethanol tolerance
Preparing YEPD liquid culture medium, sterilizing, and adding anhydrous ethanol (v/v) with different contents of 0%, 2%, 4%, 6%, 8%, 10%, and 12%. Inoculating the saccharomyces cerevisiae PLBS011 strain into YEPD culture media with different ethanol concentrations, culturing for 72h, and counting by using a blood counting plate to determine the colony number of the yeast. Ethanol tolerance was determined according to the number of yeast colonies (see FIG. 4).
From the ethanol tolerance curve, the number of live yeast bacteria decreased with the increase of ethanol content. The maximum ethanol inhibition concentration of the saccharomyces cerevisiae PLBS011 strain is 9% vol, and when the ethanol concentration is more than 9% vol, the viable yeast count is 0.
Ethanol is an inhibitor of toxin and enzyme to yeast as an end product of anaerobic fermentation, and a high-concentration substrate correspondingly enables the yeast to generate high-concentration alcohol, so that when the concentration reaches a certain value, the ethanol can inhibit the growth of the yeast, reduce the survival rate of cells and influence the activity of the cells.
Determination of acid resistance
The saccharomyces cerevisiae PLBS011 strain is inoculated into liquid culture media with different initial pH values (pH2.0, 3.0, 4.0, 5.0, 6.0 and 7.0), cultured for 20-30 h, and the OD value of the bacterial liquid is measured by a spectrophotometer at 660nm (see figure 5).
According to the acid resistance curve, the growth condition of the yeast strain is better along with the increase of the pH value within the range of 2-7, and the growth condition tends to be stable within the range of 4-77, which indicates that the acid resistance of the saccharomyces cerevisiae PLBS011 strain is better.
Determination of temperature resistance
Preparing YEPD liquid culture medium, setting the temperature gradient of the incubator at 28 deg.C, 30 deg.C, 33 deg.C, 35 deg.C, 37 deg.C, 39 deg.C, 41 deg.C, 43 deg.C, sterilizing the liquid culture medium, inoculating Saccharomyces cerevisiae PLBS011 strain, and culturing to calculate viable count (see FIG. 6).
From the temperature resistance graph, the yeast number gradually decreases with increasing temperature. The maximum inhibition value of the saccharomyces cerevisiae PLBS011 strain is 42 ℃, which shows that the temperature resistance of the saccharomyces cerevisiae PLBS011 strain is good.
The saccharomyces cerevisiae PLBS011 has excellent fermentation performance and better ethanol tolerance, acid resistance and temperature resistance, so the saccharomyces cerevisiae PLBS011 is applied to the feed for the lactating calves. In order to verify the application value of the saccharomyces cerevisiae PLBS011 in the feed for the lactating calves, the invention also provides an application method of the saccharomyces cerevisiae in the feed for the lactating calves.
First, experimental animal and raising management
Chinese Holstein calves with similar body weight and healthy bodies are selected, and lactation calves are randomly divided into 4 groups (a control group and three test groups). The lactating calf is fed with cow milk twice a day at 7 am and 4 pm, respectively, and daily ration is supplemented for 7 days. It was allowed to eat freely and guaranteed sufficient drinking throughout the test period. The starter food and the alfalfa are added according to the feed intake of the previous day, so that the residual food is ensured every time. The test period was 42 days.
The basic daily ration and the nutrition level are subject to the requirements of a calf farm.
Second, feed formula
The saccharomyces cerevisiae PLBS011 disclosed by the invention is added into basic daily ration of lactating calves, and the addition amount is 1-5%. The effective viable count of the saccharomyces cerevisiae PLBS011 is 1.38 multiplied by 1010CFU/g。
Third, measuring the index
Recording the feed feeding amount and the residual material amount of the calves every day in the test period, and calculating the daily feed intake; the weights of all lactating calves were measured with a scale before morning feeding on days 1, 21 and 42 of the test period, respectively, and the average daily gain of the calves was calculated. Nursing calves were observed daily during the test period and recorded whether they were diarrhea.
20-25 mL of blood is collected by a jugular vein of a vacuum blood collection tube before morning feeding on the 1 st day, the 21 st day and the 42 th day of the test period respectively. The collected blood samples were centrifuged, serum was separated, and serum immune index was measured.
About 200g of fresh feces samples which are not polluted by the feed are collected from each group 3 days before the test is finished, and the fresh feces samples are placed in a refrigerator at the temperature of 20 ℃ below zero for freezing and storing and are used for measuring escherichia coli and lactic acid bacteria in the feces. The Escherichia coli is cultured for 24h by adopting an eosin methylene blue culture medium, and the lactobacillus is cultured for 36h by adopting an MRS culture medium. The fecal microbial count is expressed as the logarithm of the total number of bacterial colonies contained per gram of feces [ lg (CFU/g) ].
In order to further verify the application value of the saccharomyces cerevisiae PLBS011 in the feed of the lactating calves, the lactating calves are fed by the method in the application method, the following embodiments are provided, and relevant indexes of the lactating calves in the embodiments are measured. The feeding management method of the nursing calves in each embodiment is the same, and only the addition amount of the saccharomyces cerevisiae PLBS011 in each embodiment is changed, so the same contents in each embodiment are not listed. (the addition amount of Saccharomyces cerevisiae PLBS011 in each example is shown in Table one; the results of the relevant indexes of lactating calves in each example are shown in tables two, three and four)
Comparative example
The saccharomyces cerevisiae PLBS011 of the invention is added into the conventional feed for the lactating calves, and the addition amount is 0.
After the completion of the breeding, the average daily gain of the lactating calf is measured to be 0.83kg, the average daily feed intake is measured to be 2.04kg, the material weight ratio is measured to be 2.46, and the diarrhea rate is measured to be 9.76%; measuring the content of immunoglobulin A, immunoglobulin M and immunoglobulin G in the serum of the lactating calf to be 0.69G/L, 2.47G/L and 8.90G/L respectively; the amount of Escherichia coli and the amount of lactic acid bacteria in feces of lactating calves were measured to be 5.71lg (CFU/g) and 5.71lg (CFU/g).
Example 1
The saccharomyces cerevisiae PLBS011 of the invention is added into the conventional feed for the lactating calves, and the addition amount is 2.3 percent.
After the completion of the breeding, the average daily gain of the lactating calf is 1.15kg, the average daily feed intake is 2.42kg, the material weight ratio is 2.10, and the diarrhea rate is 6.07%; measuring the content of immunoglobulin A, immunoglobulin M and immunoglobulin G in the serum of the lactating calf to be 0.95G/L, 2.85G/L and 12.55G/L respectively; the amount of Escherichia coli and the amount of lactic acid bacteria in feces of lactating calves were measured to be 5.38lg (CFU/g) and 7.12lg (CFU/g).
Example 2
The saccharomyces cerevisiae PLBS011 of the invention is added into the conventional feed for the lactating calves, and the addition amount is 3.1 percent.
After the completion of the breeding, the average daily gain of the lactating calf is 1.15kg, the average daily feed intake is 2.45kg, the material weight ratio is 2.13, and the diarrhea rate is 5.94%; measuring the content of immunoglobulin A, immunoglobulin M and immunoglobulin G in the serum of the lactating calf to be 0.89G/L, 2.90G/L and 12.62G/L respectively; the amount of Escherichia coli and the amount of lactic acid bacteria in feces of lactating calves were measured to be 5.40lg (CFU/g) and 7.09lg (CFU/g).
Example 3
The saccharomyces cerevisiae PLBS011 of the invention is added into the conventional feed for the lactating calves, and the addition amount is 3.5 percent.
After the completion of the breeding, the average daily gain of the lactating calf is 1.16kg, the average daily feed intake is 2.48kg, the material weight ratio is 2.14, and the diarrhea rate is 6.08%; measuring the content of immunoglobulin A, immunoglobulin M and immunoglobulin G in the serum of the lactating calf to be 0.93G/L, 2.93G/L and 12.59G/L respectively; the amount of Escherichia coli and the amount of lactic acid bacteria in feces of lactating calves were measured to be 5.42lg (CFU/g) and 7.10lg (CFU/g).
Because the implementation processes of the embodiments are the same, the specific processes of other embodiments are not listed, and only the three embodiments with better effects are listed.
Table one example the addition amount of saccharomyces cerevisiae PLBS 011%
Growth performance of epi-II lactating calves
Serum immunity index g/L of table three-breast-feed calves
TABLE four number of microorganisms in the feces of lactating calves lg (CFU/g)
The embodiment shows that the saccharomyces cerevisiae PLBS011 disclosed by the invention is added into basic daily ration of the lactating calf, so that the average daily gain and average daily feed intake of the lactating calf can be improved, the feed weight ratio and diarrhea rate of the lactating calf are reduced, the saccharomyces cerevisiae PLBS011 is used for promoting the growth and development of the lactating calf in the weaning period, effectively degrading anti-nutritional factors in the feed, eliminating the anti-nutritional effect of the anti-nutritional factors, and promoting the digestion and decomposition of organisms on nutrients, thereby improving the feed utilization rate. The saccharomyces cerevisiae PLBS011 is used as a feed additive and added into livestock feed, can regulate intestinal flora and enhance the digestion and absorption functions of gastrointestinal tracts, and further improves the growth performance and the feed conversion rate of animals.
The addition of the saccharomyces cerevisiae PLBS011 in the basic ration of the lactating calf can improve the content of immunoglobulin A, immunoglobulin M and immunoglobulin G in the serum of the lactating calf, and the saccharomyces cerevisiae PLBS011 can improve the secretion of immunoglobulin in the body of the lactating calf, so that the disease resistance of the body of the lactating calf is enhanced due to the antibacterial and antiviral functions of the immunoglobulin. The saccharomyces cerevisiae PLBS011 is used as a feed additive and added into livestock feed, and can be used as a non-specific immune factor to activate the immune system of the organism and regulate the immune function of the organism, thereby maintaining the health of animals.
The saccharomyces cerevisiae PLBS011 disclosed by the invention is added into basic daily ration of the lactating calf, so that the quantity of escherichia coli in feces of the lactating calf can be reduced, and the quantity of lactic acid bacteria in the feces can be increased; since escherichia coli is one of the most common and important causes of calf diarrhea, saccharomyces cerevisiae PLBS011 reduces the number of escherichia coli, thereby reducing the diarrhea rate of lactating calves; the increase of the content of lactic acid bacteria can cause the pH value of the intestinal tract of the calf to be reduced, so that the intestinal tract of the calf forms an acidic environment, the colonization of pathogenic bacteria is inhibited, and the diarrhea rate of the lactating calf is reduced. The saccharomyces cerevisiae PLBS011 is used as a feed additive and added into livestock feed, can play a synergistic effect with lactic acid bacteria, inhibits the field planting of escherichia coli together, improves the microbial environment of organisms and reduces the morbidity of animals.
In addition, the above examples show that when the addition amount of the saccharomyces cerevisiae PLBS011 of the present invention is 2.3 to 3.5% to the basal ration for lactating calves, the calves have the best productivity and the highest content of immunoglobulin in the body, and therefore, the optimal addition amount of the saccharomyces cerevisiae PLBS011 in the basal ration for calves is 2.3 to 3.5%.
In conclusion, the saccharomyces cerevisiae PLBS011 provided by the invention has good fermentation performance and good ethanol tolerance, acid tolerance and temperature tolerance, and the saccharomyces cerevisiae PLBS011 with a proper level is added into the daily ration of the lactating calf, so that the structure of the organism flora of the lactating calf can be improved, the immunity of the organism can be improved, the morbidity of the organism can be reduced, meanwhile, the utilization rate of the lactating calf to feed and the absorption rate of nutrient substances can be improved, the production performance of the lactating calf can be improved, and the economic benefit of breeding the lactating calf can be increased.
The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof, and it should be understood that various changes and modifications can be effected therein by one skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
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Sequence listing
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<120> saccharomyces cerevisiae and application thereof in feed for lactating calves
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Claims (9)
1. The Saccharomyces cerevisiae is Saccharomyces cerevisiae PLBS011 and is preserved in China general microbiological culture collection center (CGMCC) at 12 and 17 months in 2018, and the preservation number is CGMCC NO. 169951.
2. The saccharomyces cerevisiae according to claim 1, wherein the saccharomyces cerevisiae is taken from a process of brewing sake from rice.
3. The Saccharomyces cerevisiae of claim 1, wherein the Saccharomyces cerevisiae has a maximum inhibitory concentration of ethanol of 9% vol.
4. The Saccharomyces cerevisiae according to claim 1, wherein the Saccharomyces cerevisiae has a maximum inhibition temperature of 42 ℃.
5. The saccharomyces cerevisiae of claim 1, wherein the saccharomyces cerevisiae is added in a basal diet of lactating calves in an amount of 1-5%.
6. The saccharomyces cerevisiae of claim 5, wherein the saccharomyces cerevisiae is added in a basal diet of lactating calves in an amount of 2.3-3.5%.
7. A method for screening Saccharomyces cerevisiae according to claim 1, comprising the steps of:
step one preparation of a diluent
Saccharifying rice for 72h, sealing and fermenting for 40 days at 25-35 ℃, and diluting fermentation liquor to obtain a diluent;
step two isolation of the strains
Coating the diluent in a Bengal culture medium, culturing for 2-4 days at 25-30 ℃, selecting a single colony with typical yeast colony characteristics, and separating and purifying to obtain an isolated strain;
step three primary screen
Taking the multiple strains obtained in the second step in sequence, diluting the strains, inoculating the diluted strains on a lower layer culture medium plate of a TTC method, and covering an upper layer culture medium on the lower layer culture medium when the bacterial colony grows to 1-2 mm; continuously culturing for 2-4 h, taking out, and selecting red strains to be respectively transferred to test tube slopes according to the color development condition of bacterial colonies;
step four double sifting
And (4) inoculating the red bacterial strain selected in the step three into an YPD liquid culture medium, and continuously performing activation culture for 20-30 h at the temperature of 25-35 ℃ to obtain the re-screened bacterial strain.
8. The method for screening saccharomyces cerevisiae according to claim 7, wherein in the second step, the number of times of separation and purification of the strain is 2-3.
9. Use of saccharomyces cerevisiae according to claim 1 in feed for lactating calves.
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| CN113261616A (en) * | 2021-04-27 | 2021-08-17 | 内蒙古优然牧业有限责任公司 | Feed additive containing yucca, and preparation method and application thereof |
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| CN101463329A (en) * | 2009-01-07 | 2009-06-24 | 北京龙科方舟生物工程技术中心 | Saccharomyces cerevisiae, yeast preparation including the same, feed, and preparation and use thereof |
| CN102154138A (en) * | 2011-01-24 | 2011-08-17 | 天津科技大学 | Multi-resistant high-yield alcohol yeast mutant strain TT31 and screening method thereof |
| RU2716086C1 (en) * | 2018-11-09 | 2020-03-05 | Ооо "Рацио+" | Pre-starter feedstuff for calves with yeast strain saccharomyces cerevisiae |
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| CN101463329A (en) * | 2009-01-07 | 2009-06-24 | 北京龙科方舟生物工程技术中心 | Saccharomyces cerevisiae, yeast preparation including the same, feed, and preparation and use thereof |
| CN102154138A (en) * | 2011-01-24 | 2011-08-17 | 天津科技大学 | Multi-resistant high-yield alcohol yeast mutant strain TT31 and screening method thereof |
| RU2716086C1 (en) * | 2018-11-09 | 2020-03-05 | Ооо "Рацио+" | Pre-starter feedstuff for calves with yeast strain saccharomyces cerevisiae |
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