CN111938020A - Technical method for producing enterococcus faecalis by using distiller's grains - Google Patents
Technical method for producing enterococcus faecalis by using distiller's grains Download PDFInfo
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- CN111938020A CN111938020A CN202010848245.5A CN202010848245A CN111938020A CN 111938020 A CN111938020 A CN 111938020A CN 202010848245 A CN202010848245 A CN 202010848245A CN 111938020 A CN111938020 A CN 111938020A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention discloses a technical method for producing enterococcus faecalis by using distiller's grains, belonging to the technical field of fermentation engineering, comprising the following steps: activating strains, culturing primary seeds, pretreating white spirit vinasse, preparing a fermentation culture medium, and performing solid fermentation to obtain an enterococcus faecalis culture; the fermentation process of the invention adopts a method for producing enterococcus faecalis by solid state fermentation, and the whole fermentation process follows aseptic operation rules, so that the solid state culture production of the enterococcus faecalis realizes pure culture and large-scale production. The fermentation culture medium solves the problems of low quantity of live bacteria in fermentation by using the distiller's grains as main raw materials, easy pollution of mixed bacteria in liquid culture, low nutrient utilization rate and difficult separation of thalli and metabolites, and greatly improves the yield of enterococcus faecalis.
Description
Technical Field
The invention relates to the technical field of fermentation engineering, in particular to a technical method for producing enterococcus faecalis by utilizing white spirit vinasse.
Background
With the development of the white spirit industry, the utilization of the distiller's grains, which is a byproduct of white spirit production, has become the key point of the industry work. The solid vinasse is a main byproduct of the production of the white spirit and also a main pollutant of the production of the white spirit. Because the raw materials such as sorghum, wheat and the like are generally adopted, the vinasse has high acidity and contains abundant proteins, amino acids, vitamins and various trace elements. Practice has proved that the comprehensive utilization degree of the distiller's grains directly affects the development of enterprises, and if the treatment is not good, the pressure on the environment is huge; the level of distiller's grains processed feed is related to the implementation of national feed policy.
For a long time, the distiller's grains are mainly directly used as rural feed and play an important role in promoting the development of rural breeding and the virtuous cycle of the biologic chain. However, the nutrient structure of the fresh distillers ' grains cannot meet the requirements of scientific feeding and the requirements of scale development of the feeding industry, particularly, the fresh distillers ' grains have the water content of over 65 percent, are extremely difficult to store, are not easy to manage and are easy to mildew, and a large amount of distillers ' grains are randomly stacked to seriously pollute the environment, which troubles the development of enterprises.
In the prior art, some patents and other publications disclose that culture is prepared by fermentation of white spirit vinasse and applied to feed, but in the fermentation process, various mixed bacteria are generally adopted: for example, various mixed fermentation cultures of yeast, mould, lactobacillus, bacillus subtilis and the like are complicated and fussy in process, and because the culture conditions of various strains are inconsistent, the final culture can be obtained only by adopting different culture conditions to carry out 2-3 stages of culture; in addition, although a plurality of strains are mixed for fermentation, the content of the final viable bacteria is not high, and analysis shows that the final viable bacteria is not high in quantity because the suitable culture conditions of the plurality of strains are different greatly and part of the strains are dead due to the fact that the strains are not suitable for the environment under different culture conditions, so that the application value of the feed is still to be studied; for the reasons mentioned above, incomplete fermentation of the spent grains may also result, and the resulting increase in protein content of the fermentation culture is not very significant. Therefore, it is necessary to prepare a fermentation culture based on white spirit vinasse for efficient application to animal feed according to the special condition of vinasse acidity.
Disclosure of Invention
The invention aims to provide a technical method for producing enterococcus faecalis by using distiller's grains so as to solve the problems in the prior art.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides an enterococcus faecalis microecological preparation, which comprises the following components:
the carrier is a enterococcus faecalis culture and a dried remains of microalgae;
a film for coating the carrier;
and, a plurality of probiotics, the probiotics being located in the film layer;
wherein the film layer is formed by co-culturing the probiotics and the carrier at room temperature so that the probiotics are gathered on the surface of the carrier;
the enterococcus faecalis culture is produced by white spirit vinasse and comprises the following steps:
(1) activating strains: streaking enterococcus faecalis strain preserved at-80 deg.C on solid seed culture medium plate, and culturing at 35-40 deg.C for 20-24 hr; the strain is enterococcus faecalis;
(2) first-order seed culture: inoculating activated enterococcus faecalis into liquid seed culture medium by using inoculating loop to select three to five loops, placing in a shaking table, and shake-culturing for 20-24h to obtain first-class seed bacteria;
(3) pretreating spirit distiller grains: uniformly mixing the distiller's grains with water according to the mass ratio of 1:1-1:2, adding sodium bicarbonate to adjust the pH value to 6.4-7.0, inoculating first-class seed bacteria, fermenting raw materials, and culturing for 3-7 days to obtain pretreated distiller's grains;
(4) preparing a fermentation culture medium;
(5) solid fermentation: inoculating 8-10% of first-class seed bacteria into the fermentation culture medium, uniformly stirring, maintaining the humidity and the temperature, and culturing for 20-24h to obtain second-class seed bacteria;
(6) and (5) further fermenting and drying the secondary seed bacteria according to the step (5) to obtain the enterococcus faecalis culture.
Further, the microalgae is Platymonas, Chlorella, Chlorococcus, Ceratophyllum, Skeletonema, Isochrysis, Schizochytrium, Spirulina and/or Schizochytrium;
the probiotic bacteria are bifield bacteria, enterococcus faecalis, enterococcus faecium, enterococcus lactis, lactobacillus acidophilus, lactobacillus casei, lactococcus lactis, lactobacillus plantarum, pediococcus acidilactici, pediococcus pentosaceus, bacillus subtilis, rhodopseudomonas palustris and/or bacillus licheniformis.
Further, in the step (2), the shake culture condition is a condition of a temperature of 35-40 ℃ and 250 rpm; in the step (3), the raw material fermentation conditions are that the ventilation amount is 15-30% and the temperature is 30-35 ℃.
Further, the fermentation medium comprises the following components in parts by weight: 30-35 parts of pretreated distiller's grains, 5-10 parts of cane sugar, 5-9 parts of glucose, 1-3 parts of sodium chloride and KH2PH42-4 parts of MnSO4·H20.2-1.5 parts of O.
Further, the preparation method of the fermentation medium comprises the following steps: mixing the components of the fermentation medium, adding water according to the material-liquid ratio of 1:0.5-0.8, uniformly mixing, and sterilizing at the high temperature of 120-.
Further, in the step (5), the temperature is 35-37 ℃ and the humidity is 80-85%.
The invention also provides an application of the enterococcus microecological preparation in animal feed.
Further, 6-10 wt% of the feed is added into animal feed, and the mixture is mixed uniformly and used for feeding animals.
The invention discloses the following technical effects:
in the invention, the white spirit vinasse is used as a raw material to prepare the enterococcus faecalis through fermentation, the white spirit vinasse is a byproduct of white spirit production, the price is low, and the white spirit vinasse contains abundant proteins, amino acids, vitamins and various trace elements and is beneficial to the growth of thalli.
The fermentation process of the invention adopts the method for producing the enterococcus faecalis by solid state fermentation, and the whole fermentation process follows the aseptic operation rules, so that the solid state culture production of the enterococcus faecalis realizes pure culture and can realize pure cultureLarge-scale production. The fermentation culture medium solves the problems of low viable count in fermentation by using the distiller's grains as the main raw material, easy pollution of mixed bacteria in liquid culture, low nutrient utilization rate and difficult separation of thalli and metabolites, greatly improves the yield of enterococcus faecalis, and obtains the fermented product with the highest viable count of 7.8 multiplied by 1012cfu/g, and has high spore rate.
The biological fermentation product rich in enterococcus faecalis produced in the fermentation process is an ideal raw material for preparing a microecological preparation, and the enterococcus faecalis has spores, has the advantages of acid resistance, alkali resistance, 100 ℃ high temperature resistance, extrusion resistance and the like, can keep stability in the granulation process and the acid stomach environment, does not proliferate in intestinal tracts, and only rapidly develops and is converted into nutritive cells with a metabolism function in the upper section of the intestinal tracts. Enterococcus faecalis can acidify the intestinal tract, is beneficial to the absorption of iron, calcium, vitamin D and the like, promotes the growth of animals, shortens the growth cycle, can obviously improve the activity of protease, lipase and amylase and enhances the available variety of plant feed for animals; moreover, enterococcus faecalis belongs to nonpathogenic bacteria in facultative anaerobes, promotes the growth of anaerobic bacteria such as bifidobacteria, lactobacilli, clostridia and the like by self oxygen consumption, effectively inhibits the growth of aerobic bacteria such as enterobacteria, enterococci and the like in intestinal tracts, promotes the growth of normal flora of host intestinal tracts, keeps micro-ecological balance and improves the immunity of organisms.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
The "parts" in the present invention are all parts by mass unless otherwise specified.
The reagents and strains adopted by the invention are not conventional reagents and strains if no special description exists, and the selection of specific types of the reagents and strains does not have great influence on the invention;
the enterococcus faecalis adopted in the embodiment of the invention is purchased from China general microbiological culture Collection center, the preservation number is CGMCC No.3164, bacterial colonies are cultured for 24h at 37 ℃ on an MRS agar culture medium, and are enriched in an MRS liquid culture medium, milky bacterial colonies, thallus are oval, are extended along the chain direction, are paired or short-chain, are gram-positive, ferment glucose and produce lactic acid, and the bacterial strain is oval under a general microscope.
Example 1
An enterococcus faecalis microecological preparation comprising:
the carrier is a enterococcus faecalis culture and a dry remains of microalgae, and the mass ratio of the enterococcus faecalis culture to the dry remains of the microalgae is 1: 1;
a film for coating the carrier;
and, a plurality of probiotics, the probiotics being located in the film layer;
wherein the film layer is formed by co-culturing the probiotics and the carrier at room temperature so that the probiotics are gathered on the surface of the carrier;
the microalgae is chlorella; the probiotic is bifield's bacteria.
The enterococcus faecalis culture is produced by white spirit vinasse and comprises the following steps:
(1) activating strains: streaking enterococcus faecalis strain stored at-80 deg.C on solid seed culture medium plate, and culturing at 37 deg.C for 22 hr;
(2) first-order seed culture: selecting three to five rings of activated enterococcus faecalis by using an inoculating ring, inoculating the selected three to five rings of activated enterococcus faecalis into a liquid seed culture medium, placing the inoculated enterococcus faecalis into a shaking table, and performing shake culture for 22 hours at the temperature of 375 ℃ and the speed of 250rpm to prepare a first-class seed bacterium;
(3) pretreating spirit distiller grains: uniformly mixing the distiller's grains with water according to a mass ratio of 1:1.6, adding sodium bicarbonate to adjust the pH value to 6.8, inoculating first-class seed bacteria, keeping the ventilation volume at 23% and the temperature at 33 ℃, fermenting raw materials, and culturing for 5 days to obtain pretreated distiller's grains;
(4) preparing a fermentation medium, wherein the fermentation medium comprises the following components in parts by weight: 33 parts of pretreated distiller's grains, 8 parts of cane sugar, 7 parts of glucose, 2 parts of sodium chloride and KH2PH43 parts of MnSO4·H2O1 parts; mixing the components of the fermentation medium, adding water according to the feed-liquid ratio of 1:0.6, uniformly mixing, and sterilizing at 125 ℃ for 25min under high temperature and high pressure;
(5) solid fermentation: inoculating 9% of first-class seed bacteria into the fermentation culture medium, uniformly stirring, keeping the temperature at 36 ℃ and the humidity at 83%, and culturing for 22h to obtain second-class seed bacteria;
(6) and (5) further fermenting the secondary seed bacteria according to the step (5) to obtain the enterococcus faecalis culture.
In the above examples, the total viable count of enterococcus faecalis obtained by detecting the enterococcus faecalis culture by PCA plate detection method, and the total viable count is 7.8 × 1012cfu/g。
Example 2
An enterococcus faecalis microecological preparation comprising:
the carrier is a enterococcus faecalis culture and a dry remains of microalgae, and the mass ratio of the enterococcus faecalis culture to the dry remains of the microalgae is 2: 1;
a film for coating the carrier;
and, a plurality of probiotics, the probiotics being located in the film layer;
wherein the film layer is formed by co-culturing the probiotics and the carrier at room temperature so that the probiotics are gathered on the surface of the carrier;
the microalgae is hornbeam algae;
the probiotic bacteria are enterococcus faecium, enterococcus lactis, lactobacillus acidophilus and lactobacillus casei.
The enterococcus faecalis culture is produced by white spirit vinasse and comprises the following steps:
(1) activating strains: streaking enterococcus faecalis strain stored at-80 deg.C on solid seed culture medium plate, and culturing at 40 deg.C for 20 hr; the strain is enterococcus faecalis;
(2) first-order seed culture: inoculating activated enterococcus faecalis into liquid seed culture medium by using three to five rings selected by inoculating ring, placing in shaking table, and shake culturing at 40 deg.C and 250rpm for 20 hr to obtain first-class seed bacteria;
(3) pretreating spirit distiller grains: uniformly mixing the distiller's grains with water according to a mass ratio of 1:2, adding sodium bicarbonate to adjust the pH value to 6.4, inoculating first-class seed bacteria, keeping the ventilation volume at 30% and the temperature at 30 ℃, fermenting raw materials, and culturing for 7 days to obtain pretreated distiller's grains;
(4) preparing a fermentation medium, wherein the fermentation medium comprises the following components in parts by weight: pretreated distiller's grains30 parts of cane sugar 10 parts, 5 parts of glucose, 3 parts of sodium chloride and KH2PH42 parts of MnSO4·H21.5 parts of O; mixing the components of the fermentation medium, adding water according to the feed-liquid ratio of 1:0.5, uniformly mixing, and sterilizing at 130 ℃ for 25min under high temperature and high pressure;
(5) solid fermentation: inoculating 8% of first-class seed bacteria into the fermentation culture medium, uniformly stirring, keeping the temperature at 37 ℃ and the humidity at 80%, and culturing for 24h to obtain second-class seed bacteria;
(6) and (5) further fermenting the secondary seed bacteria according to the step (5) to obtain the enterococcus faecalis culture.
In the above examples, the total viable count of enterococcus faecalis obtained by detecting the enterococcus faecalis culture by PCA plate detection method, and the total viable count is 7.2 × 1012cfu/g。
Example 3
An enterococcus faecalis microecological preparation comprising:
the carrier is a enterococcus faecalis culture and a dry remains of microalgae, and the mass ratio of the enterococcus faecalis culture to the dry remains of the microalgae is 1: 2;
a film for coating the carrier;
and, a plurality of probiotics, the probiotics being located in the film layer;
wherein the film layer is formed by co-culturing the probiotics and the carrier at room temperature so that the probiotics are gathered on the surface of the carrier;
the microalgae are Isochrysis galbana, schizochytrium and Schizochytrium sp;
the probiotic bacteria are Pediococcus acidilactici, Pediococcus pentosaceus, Bacillus subtilis, Rhodopseudomonas palustris and Bacillus licheniformis.
The enterococcus faecalis culture is produced by white spirit vinasse and comprises the following steps:
(1) activating strains: streaking enterococcus faecalis strain stored at-80 deg.C on solid seed culture medium plate, and culturing at 35 deg.C for 24 hr; the strain is enterococcus faecalis;
(2) first-order seed culture: inoculating activated enterococcus faecalis into a liquid seed culture medium by selecting three to five rings with an inoculating ring, placing in a shaking table, and performing shake culture at 35 ℃ and 250rpm for 24h to obtain a first-class seed bacterium;
(3) pretreating spirit distiller grains: uniformly mixing the distiller's grains with water according to a mass ratio of 1:1, adding sodium bicarbonate to adjust the pH value to 7.0, inoculating first-class seed bacteria, keeping the ventilation quantity at 15% and the temperature at 35 ℃, fermenting raw materials, and culturing for 3 days to obtain pretreated distiller's grains;
(4) preparing a fermentation medium, wherein the fermentation medium comprises the following components in parts by weight: 35 parts of pretreated distiller's grains, 5 parts of cane sugar, 9 parts of glucose, 1 part of sodium chloride and KH2PH44 parts of MnSO4·H20.2 part of O; mixing the components of the fermentation medium, adding water according to the feed-liquid ratio of 1:0.8, uniformly mixing, and sterilizing at 120 ℃ for 25min under high temperature and high pressure;
(5) solid fermentation: inoculating 10% of first-class seed bacteria into the fermentation culture medium, uniformly stirring, maintaining the temperature at 35 ℃ and the humidity at 85%, and culturing for 20h to obtain second-class seed bacteria;
(6) and (5) further fermenting the secondary seed bacteria according to the step (5) to obtain the enterococcus faecalis culture.
In the above examples, the total viable count of enterococcus faecalis obtained by detecting the enterococcus faecalis culture by PCA plate detection method, and the total viable count is 9.2 × 1011cfu/g。
Experimental example 1
In the experiment, 60 goats with the same weight are selected for an experiment by a certain farmer in Baoding City in Hebei province, and are randomly divided into three equal groups, namely group A, group B and group C, wherein common feed purchased in the market is selected, the daily ration adopts a mixture of corn and straw, the common feed is added to the group A on the basis of the daily ration, the common feed for mixing 6% of the enterococcus faecalis microecological preparation prepared in the example 1 is added to the group B on the basis of the daily ration, the common feed for mixing 10% of the enterococcus faecalis microecological preparation prepared in the example 1 is added to the group C on the basis of the daily ration, and after the group C and the group B are fed for 2 months, compared with the group A, the daily feed intake of the group B and the group C is respectively increased by 0.34kg and; it was also found that the average daily gain increased 0.25 kg/head, 0.17 kg/head for groups B and C compared to group A.
Experimental example 2
3600 healthy and active AA broilers with similar body weight and age of 19 days are selected, the test broilers are randomly divided into a control group and a test group, each group is provided with 4 replicates, and the test lasts for 37 days. In the feeding test, the feeding condition of each group is consistent with that of the group with free feeding and free drinking. The management of each group is carried out according to the conventional method, the growth activity, health, disease condition and other information of the chickens are observed and recorded every day, and the chickens are weighed on empty stomach and analyzed in the test on the 37 th day (56 days old). Wherein the complete formula feed for the broiler chickens in the experimental group comprises 70% of corn, 25% of soybean meal, 5% of premix and 5% of the complete formula feed in terms of the enterococcus faecalis microecological preparation prepared in the addition example 1; the composition of the broiler complete formula feed of the control group is the same as that of the test group, but no enterococcus faecalis microecological preparation is added. The results show that the weight gain of the test group is increased by 8.72 percent (P is less than 0.01), the feed coefficient is increased by 6.21 percent (P is less than 0.05) and the survival rate is increased by 3.68 percent (P is less than 0.05) compared with the control group. The enterococcus faecalis microecological preparation prepared by the invention can obviously improve the daily gain, the feed coefficient and the survival rate of the broiler chickens.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (8)
1. An enterococcus faecalis microecological preparation, comprising:
the carrier is a enterococcus faecalis culture and a dried remains of microalgae;
a film for coating the carrier;
and, a plurality of probiotics, the probiotics being located in the film layer;
wherein the film layer is formed by co-culturing the probiotics and the carrier at room temperature so that the probiotics are gathered on the surface of the carrier;
the enterococcus faecalis culture is produced by white spirit vinasse and comprises the following steps:
(1) activating strains: streaking enterococcus faecalis strain preserved at-80 deg.C on solid seed culture medium plate, and culturing at 35-40 deg.C for 20-24 hr; the strain is enterococcus faecalis;
(2) first-order seed culture: inoculating activated enterococcus faecalis into liquid seed culture medium by using inoculating loop to select three to five loops, placing in a shaking table, and shake-culturing for 20-24h to obtain first-class seed bacteria;
(3) pretreating spirit distiller grains: uniformly mixing the distiller's grains with water according to the mass ratio of 1:1-1:2, adding sodium bicarbonate to adjust the pH value to 6.4-7.0, inoculating first-class seed bacteria, fermenting raw materials, and culturing for 3-7 days to obtain pretreated distiller's grains;
(4) preparing a fermentation culture medium;
(5) solid fermentation: inoculating 8-10% of first-class seed bacteria into the fermentation culture medium, uniformly stirring, maintaining the humidity and the temperature, and culturing for 20-24h to obtain second-class seed bacteria;
(6) and (5) further fermenting and drying the secondary seed bacteria according to the step (5) to obtain the enterococcus faecalis culture.
2. An enterococcal microecological formulation according to claim 1, wherein the microalgae is Platymonas, Chlorella, Chlorococcus, Ceratopsis, Skeletonema, Isochrysis, Schizochytrium, Spirulina and/or Schizochytrium;
the probiotic bacteria are bifield bacteria, enterococcus faecalis, enterococcus faecium, enterococcus lactis, lactobacillus acidophilus, lactobacillus casei, lactococcus lactis, lactobacillus plantarum, pediococcus acidilactici, pediococcus pentosaceus, bacillus subtilis, rhodopseudomonas palustris and/or bacillus licheniformis.
3. The enterococcus microecological preparation according to claim 1, wherein in the step (2), the shake culture condition is a condition of a temperature of 35-40 ℃ and 250 rpm; in the step (3), the raw material fermentation conditions are that the ventilation amount is 15-30% and the temperature is 30-35 ℃.
4. The enterococcus microecological formulation according to claim 1, wherein the fermentation medium comprises the following components in parts by weight: 30-35 parts of pretreated distiller's grains, 5-10 parts of cane sugar, 5-9 parts of glucose, 1-3 parts of sodium chloride and KH2PH42-4 parts of MnSO4·H20.2-1.5 parts of O.
5. The enterococcus microecological preparation according to claim 1, wherein the fermentation medium is prepared by the method comprising: mixing the components of the fermentation medium, adding water according to the material-liquid ratio of 1:0.5-0.8, uniformly mixing, and sterilizing at the high temperature of 120-.
6. The enterococcal microecological preparation according to claim 1, wherein in step (5), the temperature is 35-37 ℃ and the humidity is 80-85%.
7. Use of an enterococcus microecological formulation according to any one of claims 1 to 6 in animal feed.
8. The use according to claim 7, wherein 6-10 wt% is added to animal feed and mixed until uniform for feeding to animals.
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| IN192948B (en) * | 2000-02-02 | 2004-06-19 | Seagram Mfg Ltd | |
| JP2006333721A (en) * | 2005-05-31 | 2006-12-14 | Niigata Prefecture | Method for producing fermented sake lee food |
| CN104938778A (en) * | 2014-03-24 | 2015-09-30 | 瑞基海洋生物科技股份有限公司 | Aquatic animal feed |
| CN110447765A (en) * | 2019-08-23 | 2019-11-15 | 华中农业大学 | A kind of method and its application preparing bafillus natto culture using distillers ' grains |
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2020
- 2020-08-21 CN CN202010848245.5A patent/CN111938020A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IN192948B (en) * | 2000-02-02 | 2004-06-19 | Seagram Mfg Ltd | |
| JP2006333721A (en) * | 2005-05-31 | 2006-12-14 | Niigata Prefecture | Method for producing fermented sake lee food |
| CN104938778A (en) * | 2014-03-24 | 2015-09-30 | 瑞基海洋生物科技股份有限公司 | Aquatic animal feed |
| CN110447765A (en) * | 2019-08-23 | 2019-11-15 | 华中农业大学 | A kind of method and its application preparing bafillus natto culture using distillers ' grains |
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