CN111500521A - Method for preparing clostridium butyricum preparation rich in selenium by using inorganic selenium and application of clostridium butyricum preparation - Google Patents
Method for preparing clostridium butyricum preparation rich in selenium by using inorganic selenium and application of clostridium butyricum preparation Download PDFInfo
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- 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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
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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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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/30—Oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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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 provides a method for preparing a clostridium butyricum preparation rich in selenium by using inorganic selenium, which comprises the following steps: (1) preparing clostridium butyricum strains, clostridium butyricum culture media and sodium selenite; (2) adding sodium selenite into a clostridium butyricum liquid culture medium, dissolving, filtering and preparing to obtain a selenium-enriched culture medium; (3) activated clostridium butyricum strains are added into a selenium-rich culture medium according to the proportion of 5 to 10 percent, the activated clostridium butyricum is cultured for 24 hours in a constant temperature shaking table under the anaerobic condition of 37 ℃, the culture is centrifuged, sediment obtained by double distilled water washing is frozen and dried in vacuum, and the selenium-rich clostridium butyricum preparation is prepared. The clostridium butyricum preparation rich in selenium has the advantages of benefiting intestinal bacteria and organic selenium for animal health and production performance, improving animal health, promoting animal production performance, increasing the added value of livestock and poultry products, and the like. The clostridium butyricum preparation rich in selenium can be used as an animal feed additive and has wide application prospect in the production of laying hens, broilers and other livestock and poultry.
Description
Technical Field
The invention belongs to the technical field of microbial preparation production. In particular to a method for preparing a clostridium butyricum preparation rich in selenium by using inorganic selenium and application thereof.
Background
Numerous studies have shown that the addition of beneficial gut bacteria is beneficial to humans and animals. On one hand, beneficial intestinal bacteria can improve the flora structure in the intestinal tract, increase the abundance of beneficial bacteria and reduce the abundance of harmful bacteria and opportunistic pathogenic bacteria. On the other hand, the added beneficial intestinal bacteria or the added beneficial bacteria induced by the added beneficial intestinal bacteria can generate metabolites which can improve the intestinal structure and promote nutrient absorption. On the contrary, harmful intestinal bacteria and metabolites thereof damage the structure and function of the intestinal tract, increase bacteria and poison entering the host body, cause adverse effects such as immune and inflammatory reactions, and hindered nutrient absorption, and cause diseases of human and animals and reduce the production performance of domestic livestock and poultry.
Among the beneficial enterobacteria, clostridium butyricum is a gram-positive bacterium belonging to the genus clostridium of the family bacillaceae, having motility, is off-white and strictly anaerobic. The clostridium butyricum can promote the proliferation of beneficial bacteria in intestinal tracts and inhibit the proliferation of harmful bacteria, and the produced metabolite butyric acid can promote the expression of tight junction protein in intestinal tract cells, thereby being beneficial to the physical barrier of the intestinal tracts and preventing the harmful bacteria and the toxins thereof from entering the body. In the production of livestock and poultry, the clostridium butyricum preparation is added to promote the growth of animals, improve the feed conversion rate of the animals and obviously enhance the resistance of organisms to diseases.
Selenium is a trace element necessary for human and animals, has very wide physiological action and has obvious effects on improving immunity, regulating metabolism, preventing cancers and the like. Especially in the aspect of enhancing immunity, a proper amount of selenium can regulate the absorption of the body to vitamins and increase the generation of immune globulin of the body. Selenium also has antioxidant effect, is active center element of glutathione peroxidase, can scavenge free radicals in human body and animal body, and has antiaging effect. Selenium can also effectively increase the number of phagocytes in an animal body, influence the migration and chemotaxis of specific effector cells and the sterilization activity of the phagocytes, and regulate the functions of B lymphocytes, T lymphocytes, natural killer cells (NK) and neutrophils, thereby enhancing the cellular immunity of the animal body. Selenium deficiency is an important factor causing high incidence of cardiomyopathy, coronary heart disease, hypertension, diabetes and the like. The selenium supplement is beneficial to reducing the occurrence of various cardiovascular and cerebrovascular diseases, improving the symptoms of patients and improving the disease resistance of the patients. 72% of our country are in selenium-deficient and low-selenium ecological environments, and about seven hundred million people live in low-selenium and selenium-deficient areas. The intake of selenium in humans and animals is generally low and the ability to store selenium in humans is weak, so that it is necessary to eat high selenium content products frequently to obtain sufficient selenium. Therefore, the development of selenium-rich products is of great significance.
Selenium exists in nature in two ways: organic selenium and inorganic selenium. Inorganic selenium is commonly referred to as sodium selenite and sodium selenate, and is obtained as a by-product of deposits in metal deposits. However, inorganic selenium is not easy to be absorbed by human and animals, has high toxicity and is easy to pollute. Compared with inorganic selenium, the organic selenium has the characteristics of high use safety, low incidence of poisoning, high absorption and conversion rate of human bodies and animal bodies, higher nutritive value and the like. Organic selenium is usually present in the form of selenium nucleic acid, selenoprotein, selenium polysaccharide, selenomethionine, and the like. The method for producing the organic selenium is a cheap and effective method for converting inorganic selenium into the organic selenium in the thalli by utilizing the self rapid replication capacity of microorganisms.
In view of the numerous benefits of selenium and clostridium butyricum to animals, the production of clostridium butyricum enriched with selenium and its use as an additive in food or animal feed can provide the same product with the dual benefits of selenium and intestinal probiotic. The production of clostridium selenium-enriched butyrate is one of the research directions of future additives. The selenium-enriched bacteria produced at present mainly comprise selenium-enriched yeast, selenium-enriched lactic acid bacteria and selenium-enriched enterococcus faecalis. However, the production of clostridium selenobutyricum has not been reported.
Disclosure of Invention
The invention aims to solve the technical problem of how to produce a clostridium butyricum preparation rich in selenium by using sodium selenite and clostridium butyricum, and the selenium content in thalli reaches a certain enrichment degree. Aiming at the defect that the selenium-enriched clostridium butyricum is lack of an additive product in the current market, the invention utilizes inorganic selenium-sodium selenite to produce the selenium-enriched clostridium butyricum preparation, and fills the blank in the aspect of preparation of the selenium-enriched clostridium butyricum. The clostridium selenium-enriched butyrate preparation is added into livestock and poultry feeds such as laying hens, broilers and the like, so that the intestinal health of animals can be promoted, the production performance of the animals can be improved, the immunity can be improved, and the additional value of livestock and poultry products can be increased.
The technical scheme of the invention is as follows:
a first object of the present invention is to provide a method for preparing a clostridium butyricum preparation enriched with inorganic selenium, which comprises:
(1) preparing clostridium butyricum strains, clostridium butyricum culture media and sodium selenite; the clostridium butyricum culture medium comprises a clostridium butyricum solid culture medium and a clostridium butyricum liquid culture medium;
(2) adding sodium selenite into the clostridium butyricum liquid culture medium prepared in the step (1), dissolving, and filtering by adopting a 20-micron filter membrane to prepare a selenium-rich culture medium;
(3) activating a clostridium butyricum strain by using the clostridium butyricum solid culture medium prepared in the step (1), adding the activated clostridium butyricum strain liquid into a selenium-rich culture medium according to the volume ratio of 5-10%, culturing, centrifuging the culture, washing the obtained precipitate by double distilled water, washing off selenium attached to the surface to prepare the clostridium butyricum rich in selenium, and carrying out vacuum freeze drying to obtain the clostridium butyricum preparation rich in selenium.
Further, the clostridium butyricum culture medium in the step (1) is a reinforced clostridium culture medium (RCM culture medium);
further, the clostridium butyricum solid culture medium is prepared by mixing RCM culture medium powder, agar and distilled water according to the mass ratio of 38:15:1000 and carrying out autoclaving at 121 ℃ for 15 minutes;
the clostridium butyricum liquid culture medium is prepared by mixing RCM culture medium powder and distilled water according to the mass ratio of 38:1000 and carrying out autoclaving at 121 ℃ for 15 minutes.
Further, the sodium selenite in the step (1) is sodium selenite with purity not lower than 97%.
Further, the selenium concentration in the selenium-rich culture medium in the step (2) is 5-20 μ g/m L, preferably, the selenium concentration in the selenium-rich culture medium is 10 μ g/m L. under the selenium concentration defined by the invention, the conversion efficiency of the produced clostridium butyricum selenium-rich preparation can reach more than 65%.
Further, the activated clostridium butyricum strain in the step (3) is specifically as follows: the clostridium butyricum strain is clostridium butyricum viable bacteria dry powder, a clostridium butyricum freeze-drying tube is taken out, recovery activation is carried out according to requirements, sterile water is injected into the freeze-drying tube, bacteria suspension is absorbed and coated on a clostridium butyricum solid culture medium flat plate, the bacteria suspension is cultured for 24 hours under the anaerobic condition at 37 ℃, and after the activation is successful, passage is carried out for 1-2 times, so as to obtain the activated clostridium butyricum bacterial liquid.
Further, the culture conditions in the step (3) are that the culture is carried out for 24 hours in a constant temperature shaking table under the anaerobic condition at 37 ℃;
further, the centrifugation in the step (3) is to centrifuge the culture at 5000r/min for 15 minutes; the centrifugation and washing steps were repeated 3 times.
Further, the vacuum freeze-drying time in the step (3) is 48 hours.
The second purpose of the invention is to provide the clostridium butyricum preparation rich in selenium prepared by the method.
The third purpose of the invention is to provide the application of the clostridium selenium-enriched butyrate preparation in improving the growth performance of livestock and poultry, preferably laying hens or broilers, and/or improving the intestinal health of the livestock and poultry, and/or improving the organism immunity of the livestock and poultry and/or improving the production capacity of the livestock and poultry.
The fourth purpose of the invention is to provide the application of the clostridium butyricum rich in selenium in animal feed additives, which is characterized in that the animal feed additives are laying hen or broiler feed additives.
Compared with the prior art, the technical scheme of the invention has the following beneficial effects:
based on the dual benefits of selenium and beneficial intestinal bacteria clostridium butyricum on the health and production performance of animals, the invention utilizes inorganic selenium to develop the preparation method of the clostridium butyricum, and the conversion efficiency of selenium can reach more than 65 percent in the produced clostridium butyricum preparation rich in selenium under the selenium concentration limited by the invention. The clostridium butyricum preparation rich in selenium prepared by the method is added into livestock feed, can improve the intestinal health and the organism immunity of animals, improves the growth performance and the production capacity of livestock, can be used as a feed additive to be added into layer feed, can be used for producing selenium-enriched eggs, is beneficial to increasing the additional value of livestock products, and improves the economic benefit of feeders.
Detailed Description
Example 1 preparation of clostridium butyricum-rich preparations:
1) purchase of clostridium butyricum preparation: clostridium butyricum lyophilized powder (BNCC337239) was purchased from North Na Chuanglian Biotech Co.
2) The method for purchasing and preparing the clostridium butyricum culture medium comprises the steps of purchasing reinforced clostridium butyricum culture medium (RCM) (components: 10g of peptone, 10g of beef extract powder, 3g of yeast extract powder, 5g of glucose, 1g of soluble starch, 5g of sodium chloride, 3g of sodium acetate, 0.5g of L-cysteine hydrochloride and 0.5g of agar) from Hippobo biotechnology Limited of Hippocampus, dissolving 38g of RCM culture medium powder in 1000m L of distilled water, and carrying out autoclaving at 121 ℃ for 15 minutes to prepare the clostridium butyricum liquid culture medium for later use to prepare the selenium-rich culture medium.
And adding 15g of agar into 38g of RCM culture medium powder, dissolving in 1000m of L distilled water, and autoclaving at 121 ℃ for 15 minutes to prepare the clostridium butyricum solid culture medium for recovery and activation of clostridium butyricum.
3) Purchasing sodium selenite: sodium selenite (content is more than or equal to 97%) is purchased from national drug group chemical reagent limited company.
4) And (2) recovering and activating clostridium butyricum, namely taking out the freeze-dried tube stored at 4 ℃, removing a label, wiping the tube wall with 75% alcohol, transferring the tube into a biological safety cabinet for operation, scribing two circles at the center of the tube body of the freeze-dried tube by using a grinding wheel, wiping the tube wall with 75% alcohol, breaking off the tube wall by hand, injecting 0.4m L sterile water into the freeze-dried tube (containing 200 mu L bacteria powder), lightly blowing, fully mixing, sucking bacterial suspension, beating the bacterial suspension onto two plates added with a solid culture medium, wherein each plate has 200 mu L, coating, putting the plates into an anaerobic culture bag, adding an anaerobic gas generation bag and an indicator, putting the plates into an electric heating constant temperature incubator at 37 ℃ for culturing for 24 hours, and transferring for 2 times after activation is successful to restore the bacteria so as to obtain activated clostridium butyricum liquid.
5) Preparation of selenium-enriched medium for 3 treatments, group A was a control group, and the liquid medium of Clostridium butyricum (normal RCM medium) prepared in step 2) was used, group B was supplemented with sodium selenite (selenium concentration in medium 5. mu.g/M L) in RCM medium, group C was supplemented with sodium selenite (selenium concentration in medium 10. mu.g/M L) in RCM medium, and after sufficient solubilization, the prepared medium was filtered with a filter having a 20. mu.M filtration membrane.
6) And (3) culturing the selenium-rich clostridium butyricum, namely adding the activated clostridium butyricum into three ABC culture media according to the proportion of 5 percent, namely inoculating clostridium butyricum bacterial liquid with the concentration of 0.75m L into a 15m L sterile tube, putting the sterile tube into an anaerobic bag, adding an anaerobic gas generation bag and an indicator, sealing, and putting the bag into a constant-temperature shaking table with the temperature of 37 ℃ for culturing for 24 hours.
7) Centrifugal washing: the bacterial cultures were centrifuged at 5000r/min for 15 minutes, the supernatant was decanted, double distilled water was added for washing, and the centrifugation and washing steps were repeated 3 times.
8) And (3) freeze drying, namely removing a tube cover from the sterile tube containing the clostridium butyricum obtained by centrifugation, sealing the tube cover by using a preservative film, puncturing the hole, putting the tube into a vacuum freeze dryer, and freeze drying the tube for 48 hours to obtain a selenium-enriched clostridium butyricum product, wherein the group B can produce 0.0458g of clostridium butyricum per 45m L culture medium, and the group C can produce 0.0432g of clostridium butyricum per 45m L culture medium.
9) The selenium-rich content of the clostridium butyricum is detected by sending the dried clostridium butyricum to Qingdao scientific quality detection Limited company, and determining the selenium content of the clostridium butyricum by adopting inductively coupled plasma mass spectrometry (ICP-MS). The selenium content of the clostridium butyricum in the B group of the clostridium butyricum-rich is 8.29 × 103Mu g/g, the selenium content in the C group selenium-enriched clostridium butyricum is 2.37 × 104Mu.g/g. By applying the scheme, the clostridium selenium-enriched butyric acid freeze-dried powder can be obtained. The conversion rate of selenium is 50.62% and 68.3%, respectively, and sodium selenite is added to make the culture mediumWhen the selenium concentration reaches 10 mu g/m L, the conversion efficiency of the produced clostridium butyricum is the best.
Example 2 research on the enhancement of growth performance and the improvement of intestinal health of livestock and poultry by using the Clostridium selenium butyrate preparation
180 chickens of Aijia chicken with 1 day age, similar weight and good health condition are selected in the test, and the selection is randomly divided into 3 treatments, wherein each treatment is 6 times repeated, and each treatment is 10 times repeated.
Group A is a control group, and is fed with basic diet without additional selenium supplement,
the B group adds clostridium butyricum and sodium selenite into the basic diet, the addition amount of the clostridium butyricum is 200mg added into each kilogram of the basic diet, the addition amount of the sodium selenite is calculated by the selenium content, the selenium content of each kilogram of the basic diet of the B group is 0.3mg,
and C, adding the C group clostridium selenium-enriched butyrate preparation prepared in the embodiment 1 into the basic diet, wherein the addition amount of the clostridium selenium-enriched butyrate preparation is 200mg added into each kilogram of the basic diet, and the clostridium selenium-enriched butyrate preparation is diluted by using white carbon black as a carrier, so that the selenium content of each kilogram of the C group of the basic diet is 0.3mg calculated by the selenium content. The test period was 21 days.
The results showed that the broiler weight in group C was 8.28% higher than that of the control group, while the broiler weight in group C was 4.00% higher than that of group B (table 1). Compared with the control group, the gene expression levels of the broiler chicken jejunum, ileum blocking protein 1(claudin-1) and zon-1 (ZO-1) in the group B and the group C are obviously improved (Table 2). The result shows that the clostridium butyricum preparation rich in selenium can promote the expression of closely-connected related genes in intestinal epithelial tissues, promote the growth of villi and improve the intestinal health of livestock and poultry, thereby improving the growth performance of broiler chickens, and the effect of improving the growth performance of animals is better than that of a single clostridium butyricum product and inorganic selenium.
TABLE 1
TABLE 2
Example 3 study on improvement of immunity of organisms by clostridium seleni-butyrate
In the experiment, 120 Hailan brown laying hens which are 40 weeks old and have similar body weight and good health condition are selected and randomly divided into 3 treatments, wherein each treatment is 8 times repeated, and each treatment is 5 times repeated.
Group A is a control group, and is fed with basic diet without additional selenium supplement,
the B group is added with clostridium butyricum and sodium selenite in the diet, the addition amount of the clostridium butyricum is 200mg added in each kilogram of basic diet, the addition amount of the sodium selenite is calculated by selenium content, the selenium content of each kilogram of the basic diet of the B group is 0.3mg,
and C, adding the C group clostridium selenium-enriched butyrate preparation prepared in the embodiment 1 into the basic diet, wherein the addition amount of the clostridium selenium-enriched butyrate preparation is 200mg added into each kilogram of the basic diet, and the clostridium selenium-enriched butyrate preparation is diluted by using white carbon black as a carrier, so that the selenium content in each kilogram of the basic diet of the C group is 0.3 mg. The test period was 30 days.
After 30 days, one chicken is repeatedly selected for blood sampling, and the contents of immunoglobulin A (IgA), immunoglobulin Y (IgY) and immunoglobulin M (IgM) are determined, so that the result shows that compared with a control group, the serum IgA content of the laying hens in the group C is improved by 3.54 percent, the IgY content is improved by 37.39 percent, and the IgM content is improved by 23.89 percent, while compared with the group B, the serum IgA content of the laying hens in the group C is improved by 2.28 percent, the IgY content is improved by 12.70 percent, and the IgM content is improved by 11.90 percent.
From these results, it can be seen that clostridium butyricum rich in selenium can increase the content of immunoglobulin in animal serum, is beneficial to improving the immunity of animals, and is obviously superior to single clostridium butyricum products and inorganic selenium in the aspect of improving the effect of partial immunoglobulin level.
TABLE 3
Example 4 study on selenium content in livestock and poultry products
In the experiment, 240 Hailan brown laying hens which are 35 weeks old and have similar body weight and good health condition are selected and randomly divided into 3 groups, each group has 8 repetitions, and each repetition has 10 chickens.
Group A is a control group, and is fed with basic diet without additional selenium-containing additive,
the group B is added with sodium selenite in the feed, the addition amount of the sodium selenite is calculated by the selenium content, the selenium content in each kilogram of basic feed of the group B is 0.3mg,
and C, adding the C group clostridium selenium-enriched butyrate preparation prepared in the embodiment 1 into the basic diet, wherein the addition amount of the clostridium selenium-enriched butyrate preparation is 400mg added into each kilogram of the basic diet, and the clostridium selenium-enriched butyrate preparation is diluted by using white carbon black as a carrier, so that the selenium content of each kilogram of the basic diet of the C group is 0.5 mg. The test period is 14 days, and 2 eggs are randomly selected for each repetition to carry out selenium content detection.
As a result, the average selenium content of the eggs in the group A is 0.195mg/kg, the average selenium content in the group B is 0.236mg/kg, and the average selenium content in the group C is 0.294 mg/kg. Compared with the group A and the group B, the selenium content of the group C is respectively improved by 21.03 percent and 24.58 percent, and the example proves that the clostridium selenium-enriched butyrate preparation can improve the selenium content in the eggs and is used for producing the selenium-enriched eggs.
Claims (10)
1. A method of making a clostridium butyricum enriched preparation using inorganic selenium, the method comprising:
(1) preparing clostridium butyricum strains, clostridium butyricum culture media and sodium selenite; the clostridium butyricum culture medium comprises a clostridium butyricum solid culture medium and a clostridium butyricum liquid culture medium
(2) Adding sodium selenite into the clostridium butyricum liquid culture medium prepared in the step (1), dissolving, and filtering by adopting a 20-micron filter membrane to prepare a selenium-rich culture medium;
(3) activating clostridium butyricum strains by using the clostridium butyricum solid culture medium prepared in the step (1), adding the activated clostridium butyricum bacterial liquid into a selenium-rich culture medium according to the volume ratio of 5-10%, culturing, centrifuging the culture, washing the obtained precipitate by double distilled water, and performing vacuum freeze drying to prepare the selenium-rich clostridium butyricum preparation.
2. The method for preparing clostridium butyricum preparations enriched with selenium by using inorganic selenium according to claim 1, wherein the clostridium butyricum culture medium in the step (1) is an RCM culture medium.
3. The method for preparing clostridium butyricum preparations rich in selenium by using inorganic selenium according to claim 1, wherein the clostridium butyricum solid culture medium in the step (1) is prepared by mixing RCM culture medium powder, agar and distilled water in a mass ratio of 38:15:1000 and autoclaving at 121 ℃ for 15 minutes; the clostridium butyricum liquid culture medium is prepared by mixing RCM culture medium powder and distilled water according to the mass ratio of 38:1000 and carrying out autoclaving at 121 ℃ for 15 minutes.
4. The method for preparing clostridium butyricum preparation rich in selenium using inorganic selenium as claimed in claim 1, wherein the selenium concentration in the selenium-enriched medium in the step (2) is 5-20 μ g/m L, preferably 10 μ g/m L.
5. The method for preparing clostridium butyricum enriched preparation using inorganic selenium as claimed in claim 1, wherein the activated clostridium butyricum strain of the step (3) is specifically: the clostridium butyricum strain is clostridium butyricum viable bacteria dry powder, a clostridium butyricum freeze-drying tube is taken out, recovery and activation are carried out, sterile water is injected into the freeze-drying tube, bacteria suspension is absorbed and coated on a clostridium butyricum solid culture medium plate, the bacteria suspension is cultured for 24 hours under the anaerobic condition at 37 ℃, and after the activation is successful, passage is carried out for 1-2 times, so as to obtain the activated clostridium butyricum bacterial liquid.
6. The method for preparing clostridium seleniebutyricum preparation using inorganic selenium as claimed in claim 1, wherein the culture condition in the step (3) is anaerobic culture at 37 ℃ for 24 hours, and the centrifugation is centrifugation of the culture at 5000r/min for 15 minutes; the centrifugation and washing steps were repeated 3 times.
7. The method for preparing clostridium butyricum preparation enriched with selenium using inorganic selenium as claimed in claim 1, wherein the vacuum freeze-drying time of the step (3) is 48 hours.
8. A clostridium butyricum preparation enriched in selenium, prepared by the method of any claim from 1 to 7.
9. The use of clostridium butyricum enriched preparation according to claim 8 for improving the growth performance of livestock and poultry and/or improving the intestinal health of livestock and poultry and/or improving the organism immunity of livestock and poultry and/or improving the production capacity of livestock and poultry.
10. The use of a clostridium butyricum enriched preparation according to claim 8 in an animal feed additive, wherein the animal feed additive is a layer chicken or broiler feed additive.
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