CN111728094A - Intestinal tract regulation type feed additive based on high-pressure dissolution of ulva polysaccharide and preparation method and application thereof - Google Patents
Intestinal tract regulation type feed additive based on high-pressure dissolution of ulva polysaccharide and preparation method and application thereof Download PDFInfo
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- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
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Images
Classifications
-
- 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
-
- 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
-
- 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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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Birds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses an intestinal tract regulation type feed additive based on high-pressure dissolution ulva polysaccharide and a preparation method and application thereof, wherein the water-soluble polysaccharide of the feed additive accounts for more than 80% of the total sugar content; the monosaccharide composition measured by pre-column derivatization chromatography is composed of rhamnose (25.0%, DW), xylose (6.0%, DW), glucuronic acid (9.0%, DW), iduronic acid (3.1%, DW), and the sulfate group content is more than 15%. The feed additive provided by the invention has the effect of regulating the intestinal health of weaned piglets, and the product is found to be capable of effectively reducing the diarrhea rate of the piglets and improving the feed-weight ratio of the piglets to a certain extent; can improve the intestinal flora structure of piglets, inhibit the proliferation of harmful intestinal bacteria such as Mollicutes _ RF9 (Mollicutes), Escherichia-Shigella, Desulfovibrio (Desulfovibrio), and the like, and promote the proliferation of bifidobacteria and Clostridium butyricum; promoting intestinal health of piglets, and relieving intestinal inflammation; can be fermented in intestinal tract to promote the production of lactic acid, acetic acid, propionic acid and total SCFA.
Description
Technical Field
The invention relates to a feed additive, in particular to an intestinal tract regulation type feed additive based on high-pressure dissolution of ulva polysaccharide, and a preparation method and application thereof.
Background
The intestinal flora plays an important role in inflammation, immunity and metabolic diseases and is closely related to the health of a host, while diet is an important factor influencing the structure of the intestinal flora, particularly functional polysaccharide can be utilized by intestinal microorganisms, and a large amount of compounds such as Short Chain Fatty Acid (SCFA) are produced through metabolic activity of the intestinal polysaccharides, so that on one hand, the functional polysaccharide can be absorbed by the host and further metabolized as an energy source, and also can adjust the pH value of the intestinal environment and maintain the form and function of the intestinal tract, or can be used as a signal molecule to directly or indirectly influence the metabolism of the organism.
Ulva lactuca L, a large-scale marine economic algae in Ulva of the genus Chlorophyta, which is widely distributed in the coastal areas of the Western Pacific ocean, is an extremely abundant resource in wild algae in China. Research shows that ulva lactuca has the effects of resisting blood coagulation, resisting virus, enhancing immunity and the like, and the main active component of the ulva lactuca is active polysaccharide. Compared with edible green algae of other species, the ulva polysaccharide has the highest sulfate group content and has wide application and development prospects.
At present, the most research is that the ulva polysaccharide is directly extracted by hot water (80-100 ℃), the time is long, the yield is low, and the sulfate group content of the extracted polysaccharide is low; in order to improve the yield of sulfated polysaccharide, acid is added to reduce pH (1-4) for extraction at present, but the sugar unit structure, particularly sulfate groups, can be damaged when the pH is too low; when acid is added for degradation, the problem of overhigh salt concentration caused by the inevitable adjustment of pH value is also solved; moreover, to obtain high yields of polysaccharide, long extraction times are still required, which causes losses of equipment (acid corrosion); there is also a method of enzyme-assisted extraction, however, the enzyme is expensive and must satisfy the conditions of the action of the enzyme such as temperature, pH, etc. and maintain the action for a long time, and the yield improvement is still limited.
Disclosure of Invention
The invention discloses an intestinal tract regulation type feed additive based on high-pressure dissolved ulva polysaccharide and a preparation method and application thereof.
The specific contents are as follows:
a preparation process of an intestinal tract regulation type feed additive based on high-pressure dissolution of ulva polysaccharide comprises the following steps:
(1) weighing quantitative ulva naturally dried in the shade, and crushing to 100-mesh and 200-mesh;
(2) adding an ammonium oxalate solution to swell the ulva lactuca, wherein the concentration of the ammonium oxalate solution is 0.5-1%, and the mass ratio of the dried ulva lactuca to the ammonium oxalate solution is 1:15-1:30, so as to obtain ulva pulp;
(3) adding 2M phosphoric acid into Ulva lactuca to adjust pH to 5-6, and treating under 0.50-0.80 MPa for 20-40min to dissolve polysaccharide in Ulva lactuca cells sufficiently to convert into water soluble polysaccharide;
(4) concentrating the treated ulva pulp at 50-60 deg.C under reduced pressure to 1/5-1/2 volume;
(5) drying the concentrated solution at 40-50 deg.C, and pulverizing to obtain intestinal tract regulating feed additive.
The invention provides an intestinal tract regulation type feed additive based on high-pressure dissolution of ulva polysaccharide, wherein the water-soluble polysaccharide of the feed additive accounts for more than 80% of the total sugar content; the monosaccharide composition measured by pre-column derivatization chromatography is composed of rhamnose (25.0%, DW), xylose (6.0%, DW), glucuronic acid (9.0%, DW), iduronic acid (3.1%, DW), and the sulfate group content is more than 15%.
The technical scheme of the invention also comprises the application of the intestinal tract regulation type feed additive based on the high-pressure dissolution of ulva polysaccharide, and the intestinal tract regulation type feed additive can be applied to the intestinal tract health regulation of weaned piglets; the additive is added in an amount of 0.1-1% of the total weight of the feed, preferably 0.5%.
Because calcium ions exist in the seaweed cell wall, sulfate radicals can be combined to generate calcium sulfate precipitation, so that the content of sulfate radicals in the extracted polysaccharide is reduced. Wherein, ammonium oxalate can chelate calcium ions, improve the extraction efficiency of polysaccharide sulfate, and protect the sulfate from being damaged in the extraction, so that the prepared ulva polysaccharide feed additive has good function of improving intestinal microecology. The addition of the phosphoric acid does not seriously affect the acid-base balance, and the blending of the phosphorus element can make up the deficiency of the phosphorus element of the seaweed raw material to a certain extent. The dissolution rate of the water-soluble polysaccharide in the feed additive is greatly increased, the water-soluble polysaccharide accounts for more than 80 percent of the total sugar content, and the content of the functional sulfate group is more than 15 percent; the monosaccharide mainly consists of rhamnose (25.0%, DW), xylose (6.0%, DW), glucuronic acid (9.0%, DW) and iduronic acid (3.1%, DW) as determined by pre-column derivatization chromatography. Simple and easy operation, high efficiency, less pollution and reduced loss of sulfate radical.
The feed additive provided by the invention has the effect of regulating the intestinal health of weaned piglets, and the product is found to be capable of effectively reducing the diarrhea rate of the piglets and improving the feed-weight ratio of the piglets to a certain extent; can improve the intestinal flora structure of piglets, inhibit the proliferation of harmful intestinal bacteria such as Mollicutes _ RF9 (Mollicutes), Escherichia-Shigella, Desulfovibrio (Desulfovibrio), and the like, and promote the proliferation of bifidobacteria and Clostridium butyricum; promoting intestinal health of piglets, and relieving intestinal inflammation; can be fermented in intestinal tract to promote the production of lactic acid, acetic acid, propionic acid and total SCFA.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the invention and together with the description, serve to explain the principles of the invention.
FIG. 1 shows the molecular weight distribution of an intestinal tract-regulating feed additive based on high-pressure dissolution of ulva polysaccharide;
FIG. 2 is an infrared spectrogram of the intestinal tract regulation type feed additive based on high-pressure dissolution of ulva polysaccharide;
figures 3 to 5 are graphs for evaluating the intestinal tract probiotic effect of the intestinal tract regulating type feed additive based on high-pressure dissolution ulva polysaccharide on piglets;
FIG. 6 is a comparative picture of intestinal wall tissues of piglets after applying the invention;
FIG. 7 is a graph showing the effect of promoting the production of short-chain fatty acids in the intestines of piglets according to the present invention.
Detailed Description
Reference will now be made in detail to the exemplary embodiments, examples of which are illustrated in the accompanying drawings. When the following description refers to the accompanying drawings, like numbers in different drawings represent the same or similar elements unless otherwise indicated. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present invention. Rather, they are merely examples of apparatus and methods consistent with certain aspects of the invention, as detailed in the appended claims.
The invention discloses an intestinal tract regulation type feed additive based on high-pressure dissolution of ulva polysaccharide and a preparation method and application thereof, and the intestinal tract regulation type feed additive specifically comprises the following contents:
a preparation process of an intestinal tract regulation type feed additive based on high-pressure dissolution of ulva polysaccharide comprises the following steps:
(1) weighing quantitative ulva naturally dried in the shade, and crushing to 100-mesh and 200-mesh;
(2) adding an ammonium oxalate solution to swell the ulva lactuca, wherein the concentration of the ammonium oxalate solution is 0.5-1%, and the mass ratio of the dried ulva lactuca to the ammonium oxalate solution is 1:15-1:30, so as to obtain ulva pulp;
(3) adding 2M phosphoric acid into Ulva lactuca to adjust pH to 5-6, and treating under 0.50-0.80 MPa for 20-40min to dissolve polysaccharide in Ulva lactuca cells sufficiently to convert into water soluble polysaccharide;
(4) concentrating the treated ulva pulp at 50-60 deg.C under reduced pressure to 1/3 volume;
(5) drying the above concentrated solution at 40-50 deg.C, and pulverizing.
The intestinal tract regulation type feed additive based on the ulva polysaccharide dissolved out under high pressure is prepared, and the water-soluble polysaccharide accounts for more than 80% of the total sugar content; the monosaccharide composition measured by pre-column derivatization chromatography is composed of rhamnose (25.0%, DW), xylose (6.0%, DW), glucuronic acid (9.0%, DW), iduronic acid (3.1%, DW), and the sulfate group content is more than 15%.
Referring to the attached figure 1, the molecular weight distribution of the intestinal tract regulation type feed additive based on the high-pressure dissolution ulva polysaccharide prepared in the above example is determined:
the dried sample was reconstituted to 5mg/mL with distilled water and the molecular weight distribution was determined by high performance liquid chromatography. The molecular weight measurement shows that the ulva polysaccharide in the product can be degraded to a certain degree while being dissolved out, and the product mainly comprises 46469Da and 281Da components. The molecular weight of the natural algal polysaccharide is very high, so that the natural algal polysaccharide is difficult to absorb and utilize by animal bodies, the exertion of the biological activity is limited, and the molecular weight of the product in the ulva high-pressure extract is low, so that the natural algal polysaccharide can be better absorbed by animals and can exert physiological functions.
The specific operating conditions were as follows:
liquid phase apparatus: agilent 1260 high performance liquid chromatograph; a differential refractive detector;
type of liquid phase column: TSK-GEL G4000 PWxl;
liquid phase conditions: 0.2 mol/L sodium nitrate solution and 0.01 mol/L sodium dihydrogen phosphate solution are mobile phases; the column temperature is 30 ℃; the flow rate is 0.3 mL/min; the sample size is 10 muL; the differential temperature was 35 ℃.
FIG. 2 is an infrared spectrogram of the intestinal tract-regulating feed additive based on high-pressure-dissolved ulvan prepared in the above example.
Fourier transform infrared (FT-IR) spectroscopy is used here: a sample of the product (2 mg) was mixed with KBr powder and pressed into a sheet. Scanning wavelength 4000-
At wavelength, resolution 4
. The instrument comprises the following steps: nexus 470 infrared spectrometer (Nichlet instruments, Madison, Wis., USA). At 1261.46
The stretching vibration with the absorption peak S = O shows that the product contains sulfate radicals, and the absorption peak is 845.62
Is the characteristic frequency of C-S-O oscillations, indicating that the sulfate group is located in the equatorial position of the monosaccharide residue.
The invention relates to an animal experiment design of a growth promoting effect of weaned piglets, which comprises the following steps: selecting 60 weaned piglets, dividing into A, B, C, D groups with 15 weaned piglets, wherein A and B are blank groups, C and D are experimental groups, and the total period of the experiment is 30 days. Wherein the blank group is fed with common daily ration, and the experimental group is added with 0.5 percent of the product of the invention as a growth-promoting feed additive. The remaining ingredients were unchanged. The product is proved to be capable of effectively reducing the diarrhea rate of piglets and improving the feed-weight ratio of piglets to a certain extent. Experimental comparative data are as follows:
the invention relates to an animal experiment design of intestinal canal probiotic effect of weaned pigs, which comprises the following steps: selecting 30 weaned piglets, dividing into 2 groups of blank group and experiment group, 15 piglets in each group, and 30 days in total. Wherein the blank control group is fed with common daily ration, and the experimental group is added with 0.5 percent of the product of the invention as the intestinal tract probiotic additive. The remaining ingredients were unchanged. And taking the feces of the piglets ending in 30 days for 16s rDNA flora diversity analysis. The research on the regulation effect of the feed additive on the intestinal flora of weaned pigs shows that the feed additive can improve the flora structure, inhibit the proliferation of harmful intestinal bacteria such as Mollicutes _ RF9, Escherichia-Shigella, Desulfovibrio and the like, and promote the proliferation of bifidobacteria and clostridium butyricum. Fig. 3 to 5 are graphs for evaluating the intestinal tract probiotic effect of the product on piglets, wherein the significant difference exists between the experimental group and the blank group, the difference level p is less than 95%, and the analytical method is willloxon rank sum test.
Slicing and analyzing small intestines of piglets for 30 days, and observing histology to find that the piglets in the blank control group have obvious colitis symptoms and have obvious inflammatory reactions such as inflammatory cell infiltration; the infiltration of inflammatory cells of the piglets by using the product is obviously reduced, and a large amount of fibroblasts are proliferated. The product is proved to be capable of effectively promoting the intestinal health of piglets and relieving intestinal inflammation. FIG. 6 is a comparative picture of intestinal wall tissues of piglets after the application of the invention.
The invention relates to an animal experimental design for promoting the generation of short-chain fatty acid (SCFA) in the intestine of a weaned pig: 8 weaned piglets are selected and divided into an experimental group and a blank group, wherein 4 weaned piglets are selected in each group, and the total period of the experiment is 30 days. Wherein the blank control group is fed with common daily ration, and the experimental group is added with 0.5 percent of the product of the invention as the intestinal tract probiotic additive. The remaining ingredients were unchanged. The 30-day feces were taken for SCFA analysis. The composition of short-chain fatty acid products in fermentation liquor is analyzed by using an Agilent 1260 high performance liquid chromatograph, a matched ultraviolet detector and a Japanese Shodex KC-811 organic acid column, and the product fermentation is proved to be capable of promoting the generation of lactic acid, acetic acid, propionic acid and total SCFA. FIG. 7 is a graph showing the effect of the product on promoting the production of short-chain fatty acids in the intestines of piglets, wherein Nos. 1-4 represent four groups of experimental piglets; EG represents experimental group and CG represents blank group.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill 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 such modifications or substitutions do not depart from the spirit and scope of the present invention.
Claims (7)
1. A preparation process of an intestinal tract regulation type feed additive based on high-pressure dissolution of ulva polysaccharide is characterized by comprising the following steps:
(1) weighing quantitative ulva naturally dried in the shade, and crushing to 100-mesh and 200-mesh;
(2) adding an ammonium oxalate solution to swell the ulva lactuca, wherein the concentration of the ammonium oxalate solution is 0.5-1%, and the mass ratio of the dried ulva lactuca to the ammonium oxalate solution is 1:15-1:30, so as to obtain ulva pulp;
(3) adding 2M phosphoric acid into Ulva lactuca to adjust pH to 5-6, and treating under 0.50-0.80 MPa for 20-40min to dissolve polysaccharide in Ulva lactuca cells sufficiently to convert into water soluble polysaccharide;
(4) concentrating the treated ulva pulp at 50-60 deg.C under reduced pressure to 1/5-1/2 volume;
(5) drying the above concentrated solution at 40-50 deg.C, and pulverizing.
2. The process for preparing an intestinal tract regulation type feed additive based on high pressure dissolution ulvan in the claim 1, wherein the ulva pulp treated in the step (4) is concentrated to 1/3 volume by a decompression concentration scheme.
3. An enteric-modified feed additive based on high pressure-dissolved ulvan prepared by the process of claim 1, wherein the water-soluble polysaccharide accounts for more than 80% of the total sugar content; monosaccharide composition is rhamnose (25.0%, DW), xylose (6.0%, DW), glucuronic acid (9.0%, DW), iduronic acid (3.1%, DW); the content of sulfate groups is more than 15 percent.
4. The use of an enteric-modified feed supplement based on autoclaved ulvan produced according to the process of claim 1, wherein the feed supplement is used for feeding weaned piglets.
5. The use of the autoclaved ulva polysaccharide-based intestine-regulated feed additive according to claim 4, is characterized in that the diarrhea rate of weaned pigs can be effectively reduced, and the feed weight ratio of piglets can be improved to some extent; can improve the intestinal flora structure of piglets, inhibit the proliferation of harmful intestinal bacteria such as Mollicutes _ RF9 (Mollicutes), Escherichia-Shigella, Desulfovibrio (Desulfovibrio), and the like, and promote the proliferation of bifidobacteria and Clostridium butyricum; promoting intestinal health of piglets, and relieving intestinal inflammation; can be fermented in intestinal tract to promote the production of lactic acid, acetic acid, propionic acid and total SCFA.
6. The use of the enteric-coated feed additive based on the autoclaved ulvan in claim 4, wherein the feed additive is added in an amount of 0.1-1% by weight based on the total weight of the feed.
7. The use of an enteric-coated feed supplement based on autoclaved ulvan in accordance with claim 6, wherein the feed supplement is added in an amount of 0.5% by weight based on the total weight of the feed.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106615850A (en) * | 2017-01-13 | 2017-05-10 | 山东贝瑞康生物科技有限公司 | Piglet feed additive |
| CN108634118A (en) * | 2018-03-28 | 2018-10-12 | 北京慧农生物科技有限公司 | It is a kind of improve weanling pig intestinal health feed addictive and its preparation and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106615850A (en) * | 2017-01-13 | 2017-05-10 | 山东贝瑞康生物科技有限公司 | Piglet feed additive |
| CN108634118A (en) * | 2018-03-28 | 2018-10-12 | 北京慧农生物科技有限公司 | It is a kind of improve weanling pig intestinal health feed addictive and its preparation and application |
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