CN112273552B - Feed additive and preparation method and application thereof - Google Patents
Feed additive and preparation method and application thereof Download PDFInfo
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- CN112273552B CN112273552B CN202011353765.5A CN202011353765A CN112273552B CN 112273552 B CN112273552 B CN 112273552B CN 202011353765 A CN202011353765 A CN 202011353765A CN 112273552 B CN112273552 B CN 112273552B
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
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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
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
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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
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/105—Aliphatic or alicyclic compounds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/24—Compounds of alkaline earth metals, e.g. magnesium
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/28—Silicates, e.g. perlites, zeolites or bentonites
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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Abstract
The invention provides a feed additive and a preparation method and application thereof; the feed additive comprises the following raw materials in percentage by mass: 15-30% of chitosan oligosaccharide, 5-15% of yeast metabolite, 5-15% of glucoside and 40-60% of carrier. According to the invention, by reasonably compounding the chitosan oligosaccharide, the yeast metabolite, the glucoside and the carrier, the distribution of a pig intestinal microflora and the shape of an intestinal tissue can be improved, a gastric mucosa is repaired, the absorption of nutrient substances is promoted, the conversion rate of feed is improved, the growth of harmful bacteria in the intestinal tract can be inhibited, the antibacterial and antiviral abilities are improved, and the contents of serum immune factors IgA, IgG and IgM are improved, so that the diarrhea rate of piglets after weaning is effectively reduced, and the immunity of the piglets is improved; but also can promote the intestinal absorption of the piglets, improve the feed intake of the piglets, improve the daily gain of the piglets and effectively promote the growth and development of the piglets.
Description
Technical Field
The invention relates to the technical field of feeds, in particular to a feed additive and a preparation method and application thereof.
Background
After the piglets are weaned, the fed feed is changed into a solid state from a liquid state, and intestinal villi and intestinal epithelial cells are seriously damaged, so that the diarrhea of the weaned piglets is a big problem in the pig industry. The diarrhea of the weaned piglets often causes low survival rate of the piglets, slow growth and weakened resistance, and then the piglets are infected with other diseases.
In order to prevent and control diarrhea, zinc oxide and the like are added into piglet feed in many feed enterprises at present, so that a plurality of negative effects such as heavy metal pollution, serious environmental pollution, influence on other nutrient substances, toxic effect on piglets and the like are often brought while piglet diarrhea is prevented.
The development of environment-friendly growth-promoting additives which are safe, nontoxic and residue-free, can improve the immunity of animal organisms and prevent diarrhea diseases has become a necessary trend in the development of feed industry.
Disclosure of Invention
In view of the above, the invention aims to provide a feed additive and a preparation method and application thereof, by utilizing the compounding of chitosan oligosaccharide, yeast metabolite, glucoside and carrier, the reasonable compounding can improve the distribution of pig intestinal microflora and the shape of intestinal tissue, repair gastric mucosa, promote the absorption of nutrient substances, improve the conversion rate of feed, inhibit the growth of intestinal harmful bacteria, improve the antibacterial and antiviral capacity and improve the contents of serum immune factors IgA, IgG and IgM, thereby effectively reducing the diarrhea rate of piglets after weaning and improving the immunity of the piglets; but also can promote the intestinal absorption of the piglets, improve the feed intake of the piglets, improve the daily gain of the piglets and effectively promote the growth and development of the piglets.
In order to achieve the above purpose, the invention provides the following technical scheme:
the feed additive comprises the following raw materials in percentage by mass based on the total mass of the feed additive: 15-30% of chitosan oligosaccharide, 5-15% of yeast metabolite, 5-15% of glucoside and 40-60% of carrier.
In some embodiments, the method of preparing the chitosan oligosaccharide comprises: mixing chitosan with an acidic aqueous solution, carrying out mixing reaction on the obtained mixed solution and hydrolase, carrying out first solid-liquid separation, concentrating the liquid, mixing the obtained concentrated solution with absolute ethyl alcohol, standing for precipitation, and carrying out second solid-liquid separation to obtain a solid, namely chitosan oligosaccharide;
preferably, the acidic aqueous solution is prepared by mixing an acid and water in a volume ratio of (1-1.2): (10-15) mixing and preparing; the acid is at least one of glacial acetic acid, hydrochloric acid, nitric acid and phosphoric acid;
preferably, the concentration of chitosan in the mixed solution is 3-5 wt%;
preferably, the hydrolase comprises at least one of a glycosidase, a protease, and a lipase;
preferably, the dosage of the hydrolase is 1-3% of the total mass of the mixed solution;
preferably, the method further comprises the following steps before mixing the mixture and the hydrolase for reaction: firstly, adjusting the pH value of the mixed solution to 6-7;
preferably, the reaction temperature is 25-30 ℃ and the reaction time is 20-28 h.
In some embodiments, the first solid-liquid separation and the second solid-liquid separation each comprise at least one of centrifugation and filtration; preferably, the first solid-liquid separation and the second solid-liquid separation both adopt centrifugal separation, the rotation speed of the centrifugation is 4500-;
preferably, the concentration further comprises adjusting the pH of the liquid to 8, and the concentration adopts evaporation concentration; optionally, the temperature of the evaporation concentration is 45-65 ℃, and the time is 1-2 h;
preferably, the volume ratio of the concentrated solution to the absolute ethyl alcohol is (1-1.2): (10-15), wherein the standing time is 8-10 h;
preferably, the second solid-liquid separation further comprises drying the solid;
preferably, the drying adopts a vacuum drying mode; optionally, the drying temperature is 80-90 ℃ and the drying time is 1-1.5 h; more preferably, the drying further comprises washing the solid with absolute ethanol.
In some embodiments, the yeast metabolites include metabolites of at least one of candida albicans, candida tropicalis, saccharomyces cerevisiae, and han's anomala; preferably, the yeast metabolite is candida albicans, candida tropicalis, saccharomyces cerevisiae and abnormal yeast comprising the following components in percentage by mass (20-40): (20-40): (15-30): a metabolite of the mixture of (20-30);
preferably, the method for preparing the yeast metabolite comprises the following steps: at least one of candida albicans, candida tropicalis, saccharomyces cerevisiae and abnormal yeast of hank gram is placed in a fermentation culture medium for fermentation culture, and then a product after the fermentation culture is subjected to spray drying to obtain a yeast metabolite;
preferably, the fermentation culture is performed in a sterile environment.
In some embodiments, the fermentation medium comprises: 10-20g/L of glucose, 5-10g/L of peptone, 5-8g/L of yeast powder, 1-3g/L of monopotassium phosphate, 0.5-1g/L of magnesium sulfate and 15-20g/L of agar powder;
preferably, during the fermentation culture: culturing at 25-30 deg.C and pH 7-8 under shaking at 150-;
preferably, the conditions of the spray drying include: the air inlet temperature is 100-120 ℃, the sample injection flow is 8.5-10mL/min, and the air outlet temperature is 50-60 ℃.
In some embodiments, the glucoside comprises at least one of calycosin glucoside, pinoresinol diglucoside, and ascorbic acid glucoside;
preferably, the glucoside is calycosin glucoside, pinoresinol diglucoside and ascorbic acid glucoside in a mass ratio of (30-50): (30-50): (20-40).
In some embodiments, the carrier comprises at least one of maltodextrin, medical stone, calcium carbonate, zeolite powder, and choline chloride.
The invention also provides a preparation method of the feed additive, wherein the feed additive is the feed additive; the preparation method comprises the following steps: the raw materials including the chitosan oligosaccharide, the yeast metabolite, the glucoside and the carrier are mixed evenly.
An application of the feed additive in feed.
The invention also provides a feed, the feed additive;
in some embodiments, the feed is a swine feed;
preferably, the pig feed further comprises a pig feed base;
preferably, the mass ratio of the pig feed base material to the feed additive is 1000: (1-5).
The invention has the beneficial effects that:
(1) the feed additive disclosed by the invention is reasonably compounded by using chitosan oligosaccharide, yeast metabolites, glucoside and a carrier, so that the distribution of a pig intestinal microflora and the shape of an intestinal tissue can be improved, a gastric mucosa is repaired, the absorption of nutrient substances is promoted, the conversion rate of the feed is improved, the growth of harmful bacteria in the intestinal tract can be inhibited, the antibacterial and antiviral capabilities are improved, the contents of serum immune factors IgA, IgG and IgM are improved, and therefore, the diarrhea rate of piglets after weaning is effectively reduced, and the immunity of the piglets is improved; but also can promote the intestinal absorption of the piglets, improve the feed intake of the piglets, improve the daily gain of the piglets and effectively promote the growth and development of the piglets.
(2) The feed additive has simple preparation method, safe use, no toxic or side effect and no environmental pollution when being discharged into the environment, and is a nontoxic and harmless green additive; the piglet feed additive is low in preparation cost, wide in market prospect and suitable for large-scale production.
In order to make the aforementioned and other objects, features and advantages of the invention more comprehensible, preferred embodiments accompanied with figures are described in detail below.
Detailed Description
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention.
This invention may, however, be embodied in many different forms than those specifically described herein, and it will be apparent to those skilled in the art that many more modifications are possible without departing from the spirit and scope of the invention.
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. In case of conflict, the present specification, including definitions, will control.
The terms as used herein:
"prepared from … …" is synonymous with "comprising". The terms "comprises," "comprising," "includes," "including," "has," "having," "contains," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
The conjunction "consisting of … …" excludes any unspecified elements, steps or components. If used in a claim, the phrase is intended to claim as closed, meaning that it does not contain materials other than those described, except for the conventional impurities associated therewith. When the phrase "consisting of … …" appears in a clause of the subject matter of the claims rather than immediately after the subject matter, it defines only the elements described in the clause; other elements are not excluded from the claims as a whole.
When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when the range "1 ~ 5" is disclosed, the ranges described should be construed to include the ranges "1 ~ 4", "1 ~ 3", "1 ~ 2 and 4 ~ 5", "1 ~ 3 and 5", and the like. When a range of values is described herein, unless otherwise specified, the range is intended to include the endpoints thereof, and all integers and fractions within the range.
In these examples, the parts and percentages are by mass unless otherwise indicated.
"part by mass" means a basic unit of measure indicating a mass ratio of a plurality of components, and 1 part may represent any unit mass, for example, 1g or 2.689 g. If we say that the part by mass of the component A is a part by mass and the part by mass of the component B is B part by mass, the ratio of the part by mass of the component A to the part by mass of the component B is a: b. alternatively, the mass of the A component is aK and the mass of the B component is bK (K is an arbitrary number, and represents a multiple factor). It is unmistakable that, unlike the parts by mass, the sum of the parts by mass of all the components is not limited to 100 parts.
"and/or" is used to indicate that one or both of the illustrated conditions may occur, e.g., a and/or B includes (a and B) and (a or B).
The invention provides a feed additive, which comprises the following raw materials in percentage by mass based on the total mass of the feed additive: 15-30% of chitosan oligosaccharide, 5-15% of yeast metabolite, 5-15% of glucoside and 40-60% of carrier.
The preparation method of the chitosan oligosaccharide comprises the following steps: mixing chitosan with an acidic aqueous solution to obtain a mixed solution, wherein the concentration of chitosan in the mixed solution is 3-5 wt%; mixing the mixture with hydrolase to selectively cut off chitosan molecules β -a 1, 4 glycosidic bond,thereby preparing the specific low molecular weight chitosan. And then carrying out first solid-liquid separation, taking the liquid for concentration, mixing the obtained concentrated solution with absolute ethyl alcohol, standing for precipitation, and carrying out second solid-liquid separation to obtain the solid chitosan oligosaccharide.
Compared with the existing commercially available chitosan oligosaccharide, the chitosan oligosaccharide has the advantages that the effective structure is scientifically adjusted, the components of chitosan 2 and chitosan 3 are reduced, chitosan 4, chitosan 5, chitosan 6, chitosan 7 and chitosan 8 are added, and the content of chitosan 4, chitosan 5, chitosan 6, chitosan 7 and chitosan 8 is high; the chitosan oligosaccharide has 70% of effective components, and has effective structures of chitosan 4 sugar, chitosan 5 sugar, chitosan 6 sugar, chitosan 7 sugar and chitosan 8 sugar. The chitosan 4, chitosan 5, chitosan 6, chitosan 7 and chitosan 8 have the effects of proliferating beneficial intestinal flora, recovering liver cell function, enhancing immunity, resisting bacteria and tumors, regulating blood lipid, blood sugar and blood lipid levels, etc.
The acidic aqueous solution is prepared by mixing acid and water at a volume ratio of (1-1.2): (10-15) mixing and preparing; preferably, the acid comprises glacial acetic acid.
The hydrolase may be at least one of a glycosidase, a protease and a lipase; the dosage of the hydrolase is 1-3% of the total mass of the mixed solution.
Preferably, the method further comprises, before the mixing reaction of the mixture with the hydrolase: firstly, adjusting the pH value of the mixed solution to 6-7; preferably, the temperature of the reaction is 25-30 ℃ and the time is 20-28 h.
Further, the first solid-liquid separation and the second solid-liquid separation each include at least one of centrifugal separation and filtration; the first solid-liquid separation and the second solid-liquid separation both preferably adopt centrifugal separation, and optionally, the rotating speed of the centrifugation is 4500-.
Preferably, the method further comprises adjusting the pH of the liquid obtained by the first solid-liquid separation to 8 before the concentration; the concentration adopts an evaporation concentration mode. Optionally, the temperature for evaporation concentration is 45-65 deg.C, and the time is 1-2 h.
Preferably, the volume ratio of the concentrated solution to the absolute ethyl alcohol is (1-1.2): (10-15), and the standing time is 8-10 h.
Preferably, the second solid-liquid separation further comprises drying the solid obtained by the solid-liquid separation; the drying is preferably vacuum drying, optionally, the temperature of the vacuum drying is 80-90 ℃, the time is 1-1.5h, new impurities can be avoided, and the drying is more sufficient; more preferably, the drying process further comprises washing the solid with absolute ethyl alcohol, and the washing can be carried out for multiple times so as to further remove impurities on the surface of the solid.
The chitosan oligosaccharide can improve the distribution of intestinal microflora, promote the growth and reproduction of bifidobacterium, improve the immunity of organisms, reduce the pH value in intestinal tracts, inhibit the growth of harmful bacteria in the intestinal tracts, generate B vitamins, decompose carcinogens, improve the tissue morphology of the intestinal tracts, increase the density of ileum microvilli, promote the intestinal peristalsis, protect and repair gastric mucosa and improve the structure of small intestines, thereby promoting the absorption of nutrients such as protein, iron ions, zinc ions, calcium ions and the like and improving the utilization rate of feed; the chitosan oligosaccharide also has effects of stimulating organism, activating T lymphocyte and macrophage, enhancing immunity, regulating blood sugar, controlling diabetic complication, preventing cancer, protecting liver, and regulating immunity.
The yeast metabolite comprises at least one yeast metabolite of Candida albicans, Candida tropicalis, Saccharomyces cerevisiae and abnormal yeast of Wiekeeper; preferably, the yeast metabolites are candida albicans, candida tropicalis, saccharomyces cerevisiae and abnormal yeast winkler according to the mass ratio of (20-40): (20-40): (15-30): (20-30) metabolites of the mixture.
Further, the preparation method of the yeast metabolite comprises the following steps: placing at least one of candida albicans, candida tropicalis, saccharomyces cerevisiae and abnormal yeast of Wiekwom in a fermentation culture medium for fermentation culture, and then carrying out spray drying on a product after the fermentation culture to obtain a yeast metabolite;
preferably, the fermentation culture is carried out in a sterile environment, so that other mixed bacteria are prevented from interfering and influencing the fermentation product.
Further, the fermentation medium comprises: 10-20g/L of glucose, 5-10g/L of peptone, 5-8g/L of yeast powder, 1-3g/L of monopotassium phosphate, 0.5-1g/L of magnesium sulfate and 15-20g/L of agar powder.
Preferably, during the fermentation culture: culturing at 25-30 deg.C and pH 7-8 under shaking at 150-;
the conditions for the spray drying include: the air inlet temperature is 100-120 ℃, the sample injection flow is 8.5-10mL/min, and the air outlet temperature is 50-60 ℃.
The yeast metabolites comprise amino acids, nucleotides, organic acids, sugars, esters, terpenes and the like, can promote the growth and development of animals, shorten the feeding period, increase the meat and egg quantity, improve the meat quality and the lean meat percentage, improve the glossiness of fur, enhance the disease resistance of young birds and livestock, improve the immunity and reduce the non-plague sensitivity of a herd of pigs. Moreover, extreme temperatures or processing treatments do not destroy the activity of the components of this yeast metabolite.
The glucoside comprises at least one of calycosin glucoside, pinoresinol diglucoside and ascorbic acid glucoside; the glucoside is preferably calycosin glucoside, pinoresinol diglucoside and ascorbic acid glucoside in a mass ratio of (30-50): (30-50): (20-40).
It is noted that each molecule of the glucoside has two polar groups of hydrophilic group and hydrophobic group, which can increase the permeability of bacteria and virus envelope, and has antibacterial and antiviral functions, and simultaneously can synergistically improve the effects of chemicals, additives and Chinese herbal medicines, and reduce the administration cost.
The carrier comprises at least one of maltodextrin, Maifanitum, calcium carbonate, zeolite powder and choline chloride.
The invention also provides a preparation method of the feed additive, which comprises the following steps: the raw materials including the chitosan oligosaccharide, the yeast metabolite, the glucoside and the carrier are mixed evenly.
The invention also provides a feed which comprises the feed additive.
The feed can be exemplified by pig feed; the pig feed also comprises a pig feed base stock; preferably, in the pig feed: the mass ratio of the pig feed base material to the feed additive is 1000: (1-5).
The feed additive disclosed by the invention can improve the distribution of pig intestinal microflora and the form of intestinal tissues, repair gastric mucosa, promote the absorption of nutrient substances, improve the conversion rate of the feed, inhibit the growth of intestinal harmful bacteria, improve the antibacterial and antiviral capabilities and improve the contents of serum immune factors IgA, IgG and IgM by reasonably compounding the chitosan oligosaccharide, the yeast metabolite, the glucoside and the carrier, so that the diarrhea rate of the weaned piglets is effectively reduced, and the immunity of the piglets is improved; but also can promote the intestinal absorption of the piglets, improve the feed intake of the piglets, improve the daily gain of the piglets and effectively promote the growth and development of the piglets.
The preparation method of the feed additive is simple, the feed additive is safe to use, no toxic or side effect is generated, no environmental pollution is caused when the feed additive is discharged into the environment, and the feed additive is a non-toxic and harmless green additive; the piglet feed additive is low in preparation cost, wide in market prospect and suitable for large-scale production.
Embodiments of the present invention will be described in detail below with reference to specific examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Example 1
(1) Dissolving chitosan in glacial acetic acid water solution (the volume ratio of glacial acetic acid to water is 1: 10) to prepare mixed solution with the chitosan concentration of 3 wt%, adjusting the pH value of the mixed solution to 6, adding protease accounting for 1% of the total mass of the mixed solution, stirring and uniformly mixing, preserving heat for 24h under the condition of 30 ℃ water bath, then centrifuging for 30min at the rotating speed of 5000rpm, taking supernate, adjusting the pH value to 8, evaporating and concentrating for 1h at the temperature of 65 ℃, slowly adding absolute ethyl alcohol (the volume ratio of the concentrated solution to the absolute ethyl alcohol is 1: 10) into the obtained concentrated solution, standing for 8h, centrifuging again, collecting precipitate, repeatedly washing the precipitate for 3 times by using the absolute ethyl alcohol, and finally vacuum drying for 1h at the temperature of 90 ℃ to obtain the modified lower shell oligosaccharide.
(2) Candida albicans, Candida tropicalis, Saccharomyces cerevisiae and abnormal yeast Vuillemin are mixed according to the mass ratio of 20: 20: 30: 30, and placing the mixture into a fermentation medium which is aerated with sterile air and is shaken at the frequency of 150rpm for 24 hours, wherein the fermentation medium comprises: 10g/L glucose, 10g/L peptone, 5g/L yeast powder, 3g/L potassium dihydrogen phosphate, 0.5g/L magnesium sulfate and 20g/L agar powder; the temperature is controlled to be 30 ℃ and the pH value is 7 in the fermentation culture process; and finally, carrying out spray drying on the product after fermentation culture (the air inlet temperature is 100 ℃, the sample injection flow is 8.5mL/min, and the air outlet temperature is 50 ℃) to obtain a yeast product.
(3) The preparation method comprises the following steps of mixing calycosin glucoside, pinoresinol diglucoside and ascorbic acid glucoside in a mass ratio of 30: 30: 40, and uniformly mixing to obtain the glucoside.
(4) And (3) putting 30 parts of the chitosan oligosaccharide obtained in the step (1), 12 parts of the yeast metabolite obtained in the step (2), 12 parts of the glucoside obtained in the step (3) and 46 parts of maltodextrin in a mixer, and uniformly mixing to obtain the feed additive.
Example 2
(1) Dissolving chitosan in glacial acetic acid water solution (the volume ratio of glacial acetic acid to water is 1: 10) to prepare mixed solution with the concentration of chitosan being 5 wt%, adjusting the pH value of the mixed solution to 7, adding lipase accounting for 1% of the total mass of the mixed solution, stirring and uniformly mixing, keeping the temperature for 24h under the condition of water bath at 30 ℃, then centrifuging for 30min at the rotating speed of 5000rpm, taking supernate to adjust the pH value to 8, evaporating and concentrating at the temperature of 55 ℃ for 1.5h, then slowly adding absolute ethyl alcohol (the volume ratio of the concentrated solution to the absolute ethyl alcohol is 1: 10) into the obtained concentrated solution, standing for 9h, then centrifuging and collecting precipitate, repeatedly washing the precipitate for 3 times by using the absolute ethyl alcohol, and finally vacuum drying at the temperature of 80 ℃ for 1h to obtain the inferior valve modified oligosaccharide.
(2) Candida albicans, Candida tropicalis, Saccharomyces cerevisiae and abnormal yeast Vuillemin are mixed according to the mass ratio of 30: 30: 20: 20, mixing, placing in a fermentation medium with sterile air, and shaking at 150rpm for 24h, wherein the fermentation medium comprises: 20g/L of glucose, 5g/L of peptone, 8g/L of yeast powder, 1g/L of monopotassium phosphate, 1g/L of magnesium sulfate and 15g/L of agar powder; the temperature is controlled to be 28 ℃ and the pH value is 8 in the fermentation culture process; and finally, carrying out spray drying on the product after fermentation culture (the air inlet temperature is 110 ℃, the sample injection flow is 9mL/min, and the air outlet temperature is 55 ℃) to obtain a yeast product.
(3) The preparation method comprises the following steps of mixing calycosin glucoside, pinoresinol diglucoside and ascorbic acid glucoside in a mass ratio of 40: 40: 20, and uniformly mixing to obtain the glucoside.
(4) And (3) putting 25 parts of the chitosan oligosaccharide obtained in the step (1), 15 parts of the yeast metabolite obtained in the step (2), 15 parts of the glucoside obtained in the step (3) and 45 parts of maltodextrin in a mixer, and uniformly mixing to obtain the feed additive.
Example 3
(1) Dissolving chitosan in glacial acetic acid water solution (the volume ratio of glacial acetic acid to water is 1: 10) to prepare mixed solution with the chitosan concentration of 4 wt%, adjusting the pH value of the mixed solution to 6.5, adding glycosidase accounting for 2% of the total mass of the mixed solution, stirring and uniformly mixing, keeping the temperature for 24h under the condition of 30 ℃ water bath, centrifuging for 30min under the rotating speed of 5000rpm, taking supernate, adjusting the pH value to 8, evaporating and concentrating for 2h at the temperature of 45 ℃, slowly adding absolute ethyl alcohol (the volume ratio of the concentrated solution to the absolute ethyl alcohol is 1: 10) into the obtained concentrated solution, standing for 10h, centrifuging again to collect precipitate, repeatedly washing the precipitate for 3 times with the absolute ethyl alcohol, and finally vacuum drying for 1h at the temperature of 90 ℃ to obtain the inferior-modified oligosaccharide.
(2) Candida albicans, Candida tropicalis, Saccharomyces cerevisiae and abnormal yeast Vuillemin are mixed according to the mass ratio of 40: 40: 15: 20, mixing, placing in a fermentation medium with sterile air, and shaking at 150rpm for 24h, wherein the fermentation medium comprises: 15g/L glucose, 8g/L peptone, 6g/L yeast powder, 2g/L potassium dihydrogen phosphate, 0.8g/L magnesium sulfate and 18g/L agar powder; the temperature is controlled to be 30 ℃ and the pH value is 7 in the fermentation culture process; and finally, carrying out spray drying on the product after fermentation culture (the air inlet temperature is 120 ℃, the sample injection flow is 10mL/min, and the air outlet temperature is 60 ℃) to obtain a yeast product.
(3) The preparation method comprises the following steps of mixing calycosin glucoside, pinoresinol diglucoside and ascorbic acid glucoside in a mass ratio of 50: 50: 30, and mixing uniformly to obtain the glucoside.
(4) And (3) taking 20 parts of the chitosan oligosaccharide obtained in the step (1), 10 parts of the yeast metabolite obtained in the step (2), 10 parts of the glucoside obtained in the step (3) and 60 parts of maltodextrin, and uniformly mixing in a mixer to obtain the feed additive.
Example 4
The present example 4 differs from the example 1 in that: step (4): "30 parts of the chitosan oligosaccharide obtained in step (1), 12 parts of the yeast metabolite obtained in step (2), 12 parts of the glucoside obtained in step (3) and 46 parts of maltodextrin" were replaced with "30 parts of the chitosan oligosaccharide obtained in step (1), 5 parts of the yeast metabolite obtained in step (2), 5 parts of the glucoside obtained in step (3) and 60 parts of maltodextrin", and the same procedure as in example 1 was repeated.
Example 5
The present embodiment 5 differs from embodiment 1 in that: step (4): "30 parts of the chitosan oligosaccharide obtained in step (1), 12 parts of the yeast metabolite obtained in step (2), 12 parts of the glucoside obtained in step (3) and 46 parts of maltodextrin" were replaced with "15 parts of the chitosan oligosaccharide obtained in step (1), 15 parts of the yeast metabolite obtained in step (2), 15 parts of the glucoside obtained in step (3) and 55 parts of maltodextrin", and the same procedure as in example 1 was repeated.
Comparative example 1
Comparative example 1 differs from example 1 in that: the procedure of example 1 was repeated except that step (1) was omitted and "chitosan oligosaccharide" in step (4) was replaced with "commercially available chitosan oligosaccharide".
Comparative example 2
Comparative example 2 differs from example 1 in that: the procedure of example 1 was repeated except that step (1) was omitted and the yeast metabolites in step (4) were discarded.
Comparative example 3
Comparative example 3 differs from example 1 in that: the same procedure as in example 1 was repeated except that the step (3) was omitted and the glucoside in the step (4) was omitted.
Comparative example 4
Comparative example 4 differs from example 1 in that: step (4): "30 parts of the chitosan oligosaccharide obtained in step (1), 12 parts of the yeast metabolite obtained in step (2), 12 parts of the glucoside obtained in step (3) and 46 parts of maltodextrin" were replaced with "10 parts of the chitosan oligosaccharide obtained in step (1), 3 parts of the yeast metabolite obtained in step (2), 22 parts of the glucoside obtained in step (3) and 65 parts of maltodextrin", and the same procedure as in example 1 was repeated.
Comparative example 5
Comparative example 5 differs from example 1 in that: step (4): "30 parts of the chitosan oligosaccharide obtained in step (1), 12 parts of the yeast metabolite obtained in step (2), 12 parts of the glucoside obtained in step (3) and 46 parts of maltodextrin" were replaced with "35 parts of the chitosan oligosaccharide obtained in step (1), 27 parts of the yeast metabolite obtained in step (2), 3 parts of the glucoside obtained in step (3) and 35 parts of maltodextrin", and the same procedure as in example 1 was repeated.
The following are tests of the influence of the feed additive of the present invention on the productivity of weaned piglets, the diarrhea rate and on serum immune factors.
Selecting 180 ternary weaned piglets (half of each male and female) with health status and weight close to 40-50 days, randomly dividing into 10 groups, namely a test group 1, a test group 2, a test group 3, a test group 4, a test group 5, a control group 1, a control group 2, a control group 3, a control group 4 and a control group 5, and each group comprises 18 weaned piglets.
Test group 1: feeding the pig feed containing the feed additive of the example 1, wherein the pig feed is obtained by mixing a pig feed base stock and the feed additive of the example 1, and the mass ratio of the pig feed base stock to the feed additive of the example 1 is 1000: 3.
test group 2: feeding the pig feed containing the feed additive of the example 2, wherein the pig feed is obtained by mixing a pig feed base stock and the feed additive of the example 2, and the mass ratio of the pig feed base stock to the feed additive of the example 2 is 1000: 3.
test group 3: feeding the pig feed containing the feed additive of the example 3, wherein the pig feed is obtained by mixing the pig feed base stock and the feed additive of the example 3, and the mass ratio of the pig feed base stock to the feed additive of the example 3 is 1000: 3.
test group 4: feeding the pig feed containing the feed additive of the example 4, wherein the pig feed is obtained by mixing the pig feed base stock and the feed additive of the example 4, and the mass ratio of the pig feed base stock to the feed additive of the example 4 is 1000: 3.
test group 5: feeding the pig feed containing the feed additive of the example 5, wherein the pig feed is obtained by mixing a pig feed base stock and the feed additive of the example 5, and the mass ratio of the pig feed base stock to the feed additive of the example 5 is 1000: 3.
control group 1: feeding the pig feed containing the feed additive of the comparative example 1, wherein the pig feed is obtained by mixing a pig feed base stock and the feed additive of the comparative example 1, and the mass ratio of the pig feed base stock to the feed additive of the example 1 is 1000: 3.
control group 2: feeding the pig feed containing the feed additive of the comparative example 2, wherein the pig feed is obtained by mixing a pig feed base stock and the feed additive of the comparative example 2, and the mass ratio of the pig feed base stock to the feed additive of the comparative example 2 is 1000: 3.
control group 3: feeding the pig feed containing the feed additive of the comparative example 3, wherein the pig feed is obtained by mixing a pig feed base material and the feed additive of the comparative example 3, and the mass ratio of the pig feed base material to the feed additive of the comparative example 3 is 1000: 3.
control group 4: feeding the pig feed containing the feed additive of the comparative example 4, wherein the pig feed is obtained by mixing a pig feed base stock and the feed additive of the comparative example 4, and the mass ratio of the pig feed base stock to the feed additive of the comparative example 4 is 1000: 3.
control group 5: feeding the pig feed containing the feed additive of the comparative example 5, wherein the pig feed is obtained by mixing a pig feed base stock and the feed additive of the comparative example 5, and the mass ratio of the pig feed base stock to the feed additive of the comparative example 5 is 1000: 3.
the pig feed base stocks used in the test groups 1-5 and the control groups 1-5 were the same, and the pre-feeding period was 7 days and the test period was 21 days. The test results are shown in tables 1 to 3 below.
TABLE 1 Effect on production Performance
| Item | Daily gain/kg | Meat ratio of materials |
| Test group 1 | 0.4±0.02b | 1.58±0.03a |
| Test group 2 | 0.39±0.01b | 1.6±0.02a |
| Test group 3 | 0.39±0.02b | 1.6±0.03a |
| Test group 4 | 0.4±0.01b | 1.59±0.03a |
| Test group 5 | 0.39±0.02b | 1.6±0.01a |
| Control group 1 | 0.34±0.02a | 1.68±0.02b |
| ControlGroup 2 | 0.34±0.01a | 1.66±0.03b |
| Control group 3 | 0.33±0.01a | 1.61±0.03b |
| Control group 4 | 0.34±0.01a | 1.67±0.04b |
| Control group 5 | 0.33±0.03a | 1.7±0.02b |
Remarking: the same letters in the same column represent no significant difference, and different letters represent significant difference (P < 0.05); tables 2 and 3 below are the same.
As can be seen from the results in Table 1 above, the daily gain of the test groups 1-5 was significantly increased (P < 0.05) and the feed-meat ratio was significantly decreased (P < 0.05) compared to the control groups 1-5.
TABLE 2 Effect on diarrhea Rate
| Item | Rate of diarrhea/%) |
| Test group 1 | 0.63±0.27a |
| Test group 2 | 0.95±0.48a |
| Test group 3 | 0.79±0.27a |
| Test group 4 | 0.63±0.27a |
| Test group 5 | 0.63±0.27a |
| Control group 1 | 1.9±0.48b |
| Control group 2 | 2.06±0.27b |
| Control group 3 | 2.22±0.27b |
| Control group 4 | 2.38±0b |
| Control group 5 | 1.9±0.48b |
As can be seen from the results in Table 2 above, the diarrhea rates in the test groups 1 to 5 were significantly reduced (P < 0.05) as compared with the control groups 1 to 5.
TABLE 3 Effect on serum immune factors
| Item | IgA(g/L) | IgG(g/L) | IgM(g/L) |
| Test group 1 | 1.29±0.15b | 30.78±1.04b | 1.56±0.05b |
| Test group 2 | 1.4±0.19b | 30.35±0.62b | 1.6±0.04b |
| Test group 3 | 1.26±0.12b | 30.76±1.22b | 1.56±0.04b |
| Test group 4 | 1.3±0.11b | 31.01±1.19b | 1.58±0.04b |
| Test group 5 | 1.31±0.14b | 31.02±1.25b | 1.57±0.05b |
| Control group 1 | 1.01±0.11a | 26.38±1.23a | 1.31±0.07a |
| Control group 2 | 0.96±0.08a | 27.97±1.46a | 1.25±0.05a |
| Control group 3 | 0.94±0.08a | 26.16±1.39a | 1.24±0.02a |
| Control group 4 | 0.95±0.09a | 26.35±2.02a | 1.25±0.09a |
| Control group 5 | 0.96±0.1a | 26.61±1.27a | 1.24±0.12a |
As can be seen from the results in Table 3 above, IgA, IgG and IgM levels in the test groups 1 to 5 were significantly higher (P < 0.05) than in the control groups 1 to 5.
In conclusion, the feed additive provided by the invention is added into the pig feed, so that the growth and development of piglets can be remarkably promoted, diarrhea is reduced, serum immune factors are improved, and the immunity of the piglets is improved.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Furthermore, those skilled in the art will appreciate that while some embodiments herein include some features included in other embodiments, rather than other features, combinations of features of different embodiments are meant to be within the scope of the invention and form different embodiments. For example, in the claims above, any of the claimed embodiments may be used in any combination. The information disclosed in this background section is only for enhancement of understanding of the general background of the invention and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.
Claims (16)
1. The feed additive is characterized by comprising the following raw materials in percentage by mass based on the total mass of the feed additive in percentage by mass: 15-30% of chitosan oligosaccharide, 5-15% of yeast metabolite, 5-15% of glucoside and 40-60% of carrier;
the preparation method of the chitosan oligosaccharide comprises the following steps: mixing chitosan with an acidic aqueous solution, carrying out mixing reaction on the obtained mixed solution and hydrolase, carrying out first solid-liquid separation, concentrating the liquid, mixing the obtained concentrated solution with absolute ethyl alcohol, standing for precipitation, and carrying out second solid-liquid separation to obtain a solid, namely chitosan oligosaccharide;
the yeast metabolite is selected from candida albicans, candida tropicalis, saccharomyces cerevisiae and abnormal yeast of Wirkinje according to the mass ratio of (20-40): (20-40): (15-30): a metabolite of the mixture of (20-30);
the preparation method of the yeast metabolite comprises the following steps: placing candida albicans, candida tropicalis, saccharomyces cerevisiae and abnormal yeast of Wiekwom in a fermentation culture medium for fermentation culture, and then carrying out spray drying on a product after the fermentation culture to obtain a yeast metabolite; the fermentation culture is carried out in a sterile environment; in the process of fermentation culture: culturing at 25-30 deg.C and pH 7-8 under shaking at 150-180rpm for 24-30 h; the conditions of the spray drying include: the air inlet temperature is 100-120 ℃, the sample injection flow is 8.5-10mL/min, and the air outlet temperature is 50-60 ℃;
the glucoside is calycosin glucoside, pinoresinol diglucoside and ascorbic acid glucoside according to the mass ratio of (30-50): (30-50): (20-40);
the carrier comprises at least one of maltodextrin, medical stone, calcium carbonate, zeolite powder and choline chloride.
2. The feed additive of claim 1 wherein the acidic aqueous solution is prepared by mixing an acid and water in a volume ratio of (1-1.2): (10-15) mixing and preparing; the acid is at least one of glacial acetic acid, hydrochloric acid, nitric acid and phosphoric acid;
the concentration of chitosan in the mixed solution is 3-5 wt%.
3. The feed additive of claim 1 wherein the hydrolase enzyme comprises at least one of a glycosidase, a protease, and a lipase;
the dosage of the hydrolase is 1-3% of the total mass of the mixed solution.
4. The feed additive of claim 1, wherein the mixture further comprises, before reacting with the hydrolytic enzyme: firstly, adjusting the pH value of the mixed solution to 6-7;
the reaction temperature is 25-30 ℃ and the reaction time is 20-28 h.
5. The feed additive of claim 1 wherein said first solid-liquid separation and said second solid-liquid separation each comprise at least one of centrifugation and filtration.
6. The feed additive as claimed in claim 5, wherein the first solid-liquid separation and the second solid-liquid separation are performed by centrifugation at 4500-5500rpm for 25-35 min.
7. The feed additive of claim 1 further comprising adjusting the pH of said liquid to 8 prior to said concentration, wherein said concentration is by evaporative concentration at a temperature of 45-65 ℃ for a period of 1-2 hours.
8. The feed additive of claim 1 wherein the volume ratio of the concentrate to the absolute ethanol is (1-1.2): (10-15), and the standing time is 8-10 h.
9. The feed additive of claim 1 further comprising drying said solids after said second solid-liquid separation.
10. The feed additive of claim 9 wherein said drying is by vacuum drying;
the drying temperature is 80-90 ℃, and the drying time is 1-1.5 h.
11. The feed additive of claim 9 further comprising washing the solids with absolute ethanol prior to said drying.
12. The feed additive of claim 1 wherein the fermentation medium comprises: 10-20g/L of glucose, 5-10g/L of peptone, 5-8g/L of yeast powder, 1-3g/L of monopotassium phosphate, 0.5-1g/L of magnesium sulfate and 15-20g/L of agar powder.
13. A process for the preparation of a feed additive, wherein the feed additive is a feed additive according to any one of claims 1 to 12; the preparation method comprises the following steps: mixing raw materials including chitosan oligosaccharide, yeast metabolite, glucoside and carrier.
14. Use of a feed additive according to any one of claims 1 to 12 in feed.
15. A feed comprising the feed additive according to any one of claims 1 to 12.
16. The feed of claim 15, wherein the feed is a pig feed, the pig feed further comprises a pig feed base, and the mass ratio of the pig feed base to the feed additive is 1000: (1-5).
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