CN116637176A - Natural livestock and poultry immunity activator and preparation method and application thereof - Google Patents
Natural livestock and poultry immunity activator and preparation method and application thereof Download PDFInfo
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- CN116637176A CN116637176A CN202310661484.3A CN202310661484A CN116637176A CN 116637176 A CN116637176 A CN 116637176A CN 202310661484 A CN202310661484 A CN 202310661484A CN 116637176 A CN116637176 A CN 116637176A
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Abstract
The invention discloses a natural livestock and poultry immune activator, which comprises an activator crude product and snailase, wherein the activator crude product is prepared from the following components: 8-15 parts of aloe skin, 3-8 parts of dried lotus root residues, 4-9 parts of pine nut shells, 10-20 parts of pteris longifolia, 5-8 parts of hemerocallis, 5-8 parts of red algae, 6-12 parts of termitomyces albuminosus, 4-9 parts of lentinus edodes and 3-6 parts of elm. Also discloses a preparation method of the livestock and poultry natural immune activator, which comprises the following steps: cleaning components in the crude product of the activating agent, drying, and crushing twice to obtain a powdery raw material; mixing the powdery raw materials, soaking the mixture in warm water overnight, leaching the mixture with hot water, and drying the obtained filter residue to obtain a product A, and carrying out ultrasonic-microwave-assisted fast extraction on the obtained filtrate to obtain a product B; mixing the product A and the product B, and then mixing with snailase to obtain the immune activator; the immune activator is added into livestock and poultry basic ration. The invention reserves the important functional polysaccharide, oligosaccharide protein and other nutrients in the raw materials to the greatest extent, and the whole production process is energy-saving and low-consumption.
Description
Technical Field
The invention belongs to the technical field of livestock and poultry immune activators, and particularly relates to a livestock and poultry natural immune activator, and a preparation method and application thereof.
Background
In the face of increasing cost pressure and price fluctuation risks, some large feed companies fight for the market rapidly, the large pens fight for the market share to ensure normal production, but under the current fully-competitive market environment, the key for promoting the steady development of the feed enterprises is to continuously break through the technical barriers of the feed from technological innovation. In the aspect of feed technology research, the improvement of the technical content of the feed formula is important, which is not only important to ensure the brand competitiveness, but also directly influences the economic benefit of feed enterprises, so that the whole feed industry is paid attention to.
Chinese patent No. 201510221310.0 discloses that the Chinese medicinal polysaccharide immunopotentiator for livestock and poultry is prepared from astragalus root, pilose asiabell root, poria cocos, mushroom in a proportion of 1:1:1:1, mainly obtained by ultrasonic cell crushing, alcohol precipitation, washing, drying and other steps, the invention can be evenly mixed into feed or dissolved in livestock and poultry drinking water for livestock and poultry, and plays a role in promoting the enhancement of nonspecific cell immune bioactivity and humoral immune response; in general, the preparation process in the patent has poor practical production feasibility, the preparation process of the traditional Chinese medicine polysaccharide immunopotentiator needs multiple times of extraction, is excessively complicated, has high energy consumption, and has limited yield of the traditional Chinese medicine polysaccharide.
The invention CN201910277425.X provides an edible fungus polysaccharide composition with immunity enhancing effect, which is composed of ganoderan and polyporus polysaccharide, and has remarkable effects of improving the activity of natural killer cells and macrophage functions of the organism and enhancing the immunity of the organism; however, the improvement effect of other nutrients and bioactive substances in other kinds of feeds on the immune function of animals is ignored in the patent, and thus the immune enhancing effect is general.
In order to solve the problems, a natural livestock and poultry immune activator and a preparation method and application thereof are provided.
Disclosure of Invention
The invention aims to solve the technical problem of providing a natural livestock and poultry immune activator, and a preparation method and application thereof. The preparation method is obtained through several important steps of shearing and crushing, hot water leaching and ultrasonic microwave assisted fast extraction, is simple and convenient to prepare, retains the nutrients of important functional polysaccharide, oligosaccharide, protein and the like in the raw materials to the greatest extent, saves energy in the whole production process, has low consumption, and has small environmental pollution and broad market prospect.
In order to solve the technical problems, the invention adopts the following technical scheme: the natural livestock and poultry immunity activator comprises a crude activator product and snailase, wherein the crude activator product is prepared from the following components in parts by weight: 8-15 parts of aloe skin, 3-8 parts of dried lotus root residues, 4-9 parts of pine nut shells, 10-20 parts of pteris longifolia, 5-8 parts of hemerocallis, 5-8 parts of red algae, 6-12 parts of termitomyces albuminosus, 4-9 parts of lentinus edodes and 3-6 parts of elm; the mass ratio of the snailase to the crude activator is 2:15.
The preparation method of the livestock and poultry natural immune activator is also provided, and comprises the following steps:
s1, cleaning each component in the crude product of the activator respectively, and then drying at a low temperature to obtain each component after drying;
s2, sequentially crushing the dried components obtained in the step S1, and then secondarily crushing to obtain nine powdery raw materials;
s3, respectively weighing the nine powdery raw materials obtained in the step S2, uniformly mixing, adding warm water for soaking overnight, adding hot water, heating the water while stirring, leaching for 2 hours after the water temperature is 45 ℃, filtering to obtain filtrate and filter residues, and drying the filtrate to obtain a product A;
s4, adding distilled water into the filter residue obtained in the S3, uniformly mixing, then placing the mixture in a multi-frequency power ultrasonic microwave synergistic combination workstation for extraction, and drying to obtain a product B;
s5, mixing the product A obtained in the step S3 with the product B obtained in the step S4 to obtain a crude product of the activator; and then uniformly mixing the crude activator product with snailase to prepare the immune activator.
Preferably, the low-temperature drying temperature in S1 is 45-57 ℃ and the time is 15h.
Preferably, the pre-crushing in S2 uses a shear crusher with a rotation speed of 3X 10 3 r/min, the time is 10-20s; the secondary crushing uses a double-hammer horizontal rail type crusher with the aperture of 7.0 mm.
Preferably, the mass ratio of the total mass of the nine powdery raw materials to the warm water in the S3 is 1:5; the volume ratio of the hot water to the warm water is 5:1; the temperature of the warm water is 25 ℃, and the temperature of the hot water is 78 ℃.
Preferably, the drying temperatures in S3 and S4 are 55-70deg.C, the drying time in S3 is 120min, and the drying time in S4 is 240min.
Preferably, the mass ratio of the filter residue to the distilled water in the step S4 is 1:2; the ultrasonic power of the multi-frequency variable power ultrasonic wave and microwave cooperative combination workstation is 150w, the microwave power is 180w, the temperature is 60 ℃, and the extraction time is 20min.
The application of the livestock and poultry natural immune activator is also provided, and the addition mass ratio of the immune activator in the livestock and poultry basic ration is 0.50-1.20%.
Compared with the prior art, the invention has the following advantages:
1. the immune activator provided by the invention consists of marine algae, medicinal fungi, several terrestrial plant wastes and snailase, and the immune additive is obtained by matching the above raw materials, has the advantages of sufficient and balanced nutrition, low cost, large gastrointestinal mucosa penetration, super-strong immune induction activity, obviously improved growth performance once the livestock and poultry orally take, and greatly reduced death and panning quantity: firstly, the immune activating additive plays a role in maintaining important immune organs of organisms, so that serious damage of harmful substances (such as heavy metal elements and mycotoxins) in the additive to the immune organs of livestock and poultry is avoided, the development perfection of immune organs such as thymus, bursa of Fabricius, spleen and the like can be positively promoted, and the generation, proliferation, differentiation and even maturation of immune cells in vivo are indirectly influenced; secondly, the immune activator can effectively promote the rapid proliferation of certain leucocytes and lymphocytes and stimulate the secretion of immune antibodies and related cytokines to increase; in addition, the immune activator can regulate intestinal microflora and maintain the normal function of intestinal mucosa immunity by reducing the intestinal firmicutes and increasing the relative abundance of the bacteroides;
the dry lotus root residue, pine nut shells and aloe barks are offcuts produced in the industrial processing process, most of the offcuts are discarded at present, so that environmental pollution and resource waste are caused, and the molecular structure of the mannans in the aloe barks has higher acetyl content, so that the immunoregulation pharmacological activity is stronger than that of other mannans; the termitomyces albuminosus is a high-protein, low-fiber and low-fat edible fungus, wherein active substances such as polysaccharide, polyphenol, saponin, cellulose and the like can timely remove excessive free radicals in the organism, delay the peroxidation process of cell lipid, maintain various immune cell normal structures and influence the relative proportion of T cell subsets in peripheral blood, in particular T lymphocyte proliferation; laccase in lentinus edodes fruiting body has strong heat resistance, can oxidize most lignin in feed into small molecule nutrition which is easy to be absorbed by animals, effectively reduces the content of anti-nutritional factors, obviously improves the level of soluble protein, further provides sufficient and various nutrition for massive secretion of immune active substances IgG, igA, igM and IL-2 and IL-Y, and has the unique advantages of no toxicity, no drug residues and no drug resistance, and has very obvious effects in activating macrophages and inducing cytotoxicity of killer macrophages and antibody-dependent macrophages; ulmus pumila, also called Ulmus pumila, belongs to the Polyporales Ulmaceae and is a plastic-rubber-plastic fungus, the Ulmus pumila protein and oligopeptide can provide energy for organisms, the elm has the effects of relieving fatigue, and the elm also contains some antibacterial active ingredients with good heat stability, such as volatile oil sesquiterpenes, phenols and organic acids; the marine algae resources in China are rich, however, the development and the utilization of the marine algae in the feed are limited at present, and the grape fern algae, the red hair algae and the hemerocallis adopted by the invention have important effects of absorbing heavy metals, and can play the active functions of immunoregulation, anti-inflammatory and anticancer by means of sulfated polysaccharide, fern phycoerythrin and various monosaccharides in the marine algae to mediate the signal path of the immunostimulatory activity; the snailase is a mixed enzyme prepared from the marbled capsule and the alimentary canal of snails, contains more than 20 enzymes such as cellulase, pectase, amylase, protease and the like, is mixed with the crude product of the immune activator according to a proportion and applied to livestock and poultry production, can rapidly degrade polysaccharide and protein which are difficult to digest by organisms in unconventional feed resources, and improves the nutrition concentration of oligosaccharide, small peptide, mannans and the like of the immune activator, thereby improving the nutrition and digestibility of products, and simultaneously promoting the absorption and conversion process of organisms to feed nutrients.
2. The invention abandons a series of complex crude polysaccharide purification processes such as protein removal, pigment degradation and the like, and furthest reserves functional polysaccharide, oligosaccharide and the like and basic units thereof in the raw materials. In the key preparation process, shearing and crushing are carried out firstly and then conventional crushing is carried out, so that the crushing time of the aloe peel and the like which are unconventional raw materials can be shortened on the basis of ensuring the required crushing fineness, and the serious pollution of abrasive metal elements of a crusher to the raw materials is reduced. In addition, compared with the conventional process, the crushing step and the following two key processes of hot water leaching and ultrasonic microwave assisted fast extraction are matched, so that the purposes of thoroughly destroying the cell walls of raw materials and promoting the extremely quick dissolution of nutrient functional substances can be achieved, and the production efficiency of the feed is greatly improved. Therefore, the preparation process disclosed by the invention is simple, convenient, efficient, energy-saving, low in consumption and pollution, and has a good batch large-scale application prospect.
3. According to the multi-frequency variable power ultrasonic microwave cooperative combination workstation, the multi-frequency transducer is utilized in the cooperative combination workstation to realize ultrasonic microwave generation characteristics of various frequencies of equipment, and the working frequency can be adjusted in real time according to the parameter requirements of different raw material components on ultrasonic microwaves, so that the multi-frequency ultrasonic microwave generation system of the combination workstation can further improve the extraction rate of effective active ingredients of raw materials.
The technical scheme of the invention is further described in detail through the drawings and the embodiments.
Drawings
FIG. 1 is jejunal mucosa tight junction protein mRNA expression in commercial pigs of example 3 and comparative example 5.
Detailed Description
Example 1
The livestock and poultry natural immune activator comprises an activator crude product and snailase, wherein the activator crude product is prepared from the following components in parts by weight: 8 parts of aloe skin, 3 parts of dried lotus root residues, 4 parts of pine nut shells, 10 parts of pteris longifolia, 5 parts of hemerocallis, 5 parts of red algae, 6 parts of termitomyces albuminosus, 4 parts of lentinus edodes and 3 parts of elm; the mass ratio of the snailase to the crude activator is 2:15.
The preparation method of the livestock and poultry natural immune activator comprises the following steps:
s1, respectively screening out impurities in nine components in the crude product of the activating agent by adopting an intelligent double-layer blast type winnowing machine, respectively cleaning, and then drying at a low temperature of 45 ℃ for 15 hours to obtain each component after drying;
s2, sequentially using a shearing pulverizer to obtain the dried components in S1 in a ratio of 3×10 3 Pre-crushing for 10s at the rotating speed of r/min, and then performing secondary crushing by using a double-hammer horizontal rail type crusher with the aperture of 7.0mm to obtain nine powdery raw materials;
s3, weighing nine powdery raw materials obtained in the step S2 according to the weight portions respectively, and uniformly mixing the nine powdery raw materials according to the mass ratio of 1:5, adding warm water with the temperature of 25 ℃ for soaking overnight (15 h), adding hot water with the temperature of 78 ℃ for the next day, stirring while heating the water, leaching for 2h after the water temperature is 45 ℃ to obtain filtrate and filter residue, and drying the filtrate at the temperature of 55 ℃ for 120min to obtain a product A; the volume ratio of the hot water to the warm water is 5:1;
s4, adding distilled water into the filter residue obtained in the step S3, uniformly mixing, placing the filter residue and the distilled water in a mass ratio of 1:2 in a multi-frequency power ultrasonic microwave synergistic combination workstation, setting the ultrasonic power to be 150w, the microwave power to be 180w, the temperature to be 60 ℃, extracting for 20min, and drying for 240min at the temperature of 55 ℃ to obtain a product B;
s5, mixing the product A obtained in the step S3 with the product B obtained in the step S4 to obtain a crude product of the activator; the crude activator and snailase were then mixed according to 15:2, mixing uniformly to obtain the immune activator.
The low temperature drying temperature in this embodiment S1 may also be 46 ℃, 47 ℃, 52 ℃, 55 ℃ or 56 ℃; the time for the pre-pulverization in S2 may also be 12S, 13S, 14S, 15S, 16S, 17S, 18S or 19S; the drying temperature in S3 can be 57 ℃, 58 ℃, 62 ℃, 65 ℃, 68 ℃ or 69 ℃; the drying temperature in S4 may also be 57 ℃, 58 ℃, 62 ℃, 65 ℃, 68 ℃ or 69 ℃.
The nutritional ingredients of the immune activator of example 1 were measured and analyzed for polysaccharide content, and the measurement results were as follows:
(1) The measurement results of the nutritional ingredients are shown in Table 1.
TABLE 1 conventional nutritional ingredients of the immune activators prepared in example 1
(2) The mass fraction of polysaccharide of the immune activator prepared in example 1 is 34.50%, the glucomannan content is 8.62%, and the specific operation steps are as follows:
1) Quantitative determination of water-soluble polysaccharides
A. Preparation of glucose standard solution: 100mg of glucose standard is weighed, and dissolved in water to prepare a glucose solution of 0.4 mg/ml. 1 part of each of 0.1, 0.2, 0.3, 0.4, 0.5 and 0.6mL is accurately removed, water is added to 2.0mL, then 1.0mL of 5% phenol water solution is added, 5.0mL of concentrated sulfuric acid is rapidly added dropwise after shaking, water bath heating is carried out at 85 ℃ for 20min after 10min, and the mixture is rapidly cooled to room temperature after being taken out. Another 2.0mL of distilled water was used as a blank.
B. Drawing a standard curve: the absorbance of the sample was measured at 490nm using an ultraviolet spectrophotometer. And (3) taking the glucose concentration as an abscissa and the absorbance value as an ordinate as a standard curve to obtain a correlation regression equation.
C. Sample content determination: 50mg of the immune activator is weighed, a sample liquid is prepared according to the method described in the A, absorbance is measured, and the relative content of polysaccharide is calculated by substituting the absorbance into a regression equation of B.
2) Glucomannan analysis
A. And (3) making a standard curve: refer to GB/T18104-2000.
B. Preparing refined powder: weighing 0.12-0.15g of immune activator, adding 50ml of 85% ethanol, keeping the temperature in 50 ℃ constant-temperature water bath for 30min, stirring, filtering, extracting with 85% ethanol twice, and steaming to remove ethanol.
C. Preparing an extracting solution: adding 60ml of distilled water into the refined powder, swelling in 35 ℃ water bath for 4 hours, continuously stirring, homogenizing for 40 seconds, adding water to fix the volume, and centrifuging to obtain 10.0ml of supernatant.
D. Preparing a hydrolysate: respectively transferring 1.0ml of the extract into 3 volumetric flasks of 50ml, adding 0.5ml of sulfuric acid solution of 3mol/L, shaking, sealing in boiling water bath, hydrolyzing for 1.5 hr, taking out, and cooling. 6mol/L sodium hydroxide solution was added thereto and shaken well.
E. Measurement of glucomannan: to the 3 parts of the above-mentioned hydrolysis solution, 1.5ml of 3, 5-dinitrosalicylic acid reagent was added, respectively. Heating in boiling water bath for 5min, cooling, metering to 50ml, colorizing at 550nm position by using a spectrophotometer, calculating glucose content by using a standard curve, wherein glucomannan content=0.9T×100/m (m is the mass of an immune activator, and T is the glucose content of hydrolysate corresponding to the standard curve).
(3) The immune activator prepared in the example 1 is subjected to detection of the contents of heavy metal elements such As mercury (Hg), cadmium (Cd), lead (Pb) and arsenic (As), and the detection results are shown in Table 2;
the quality standard of the additive in the immune activator prepared in the embodiment 1 reaches the sanitary standard (Hg is less than or equal to 0.10mg/kg, cd is less than or equal to 1.00mg/kg, pb is less than or equal to 40.0mg/kg, as is less than or equal to 10.0 mg/kg) in the national standard-natural plant feed additive general rule, and the detection method is referred to GB/T13081-2006, GB/T13082-1991, GB/T13080-1991 and GB/T13079-2006.
TABLE 2 heavy metal content of the immune activators prepared in EXAMPLE 1
Example 2
The livestock and poultry natural immune activator comprises an activator crude product and snailase, wherein the activator crude product is prepared from the following components in parts by weight: 10 parts of aloe skin, 5 parts of dried lotus root residues, 6 parts of pine nut shells, 15 parts of pteris longifolia, 6 parts of hemerocallis fulva, 6 parts of red hair algae, 9 parts of termitomyces albuminosus, 6 parts of lentinus edodes and 4 parts of elm; the mass ratio of the snailase to the crude activator is 2:15.
The preparation method of the livestock and poultry natural immune activator comprises the following steps:
s1, respectively screening out impurities in nine components in the crude product of the activating agent by adopting an intelligent double-layer blast type winnowing machine, respectively cleaning, and then drying at a low temperature of 50 ℃ for 15 hours to obtain each component after drying;
s2, sequentially using a shearing pulverizer to obtain the dried components in S1 in a ratio of 3×10 3 Pre-crushing for 20s at the rotating speed of r/min, and then performing secondary crushing by using a double-hammer horizontal rail type crusher with the aperture of 7.0mm to obtain nine powdery raw materials;
s3, weighing nine powdery raw materials obtained in the step S2 according to the weight portions respectively, and uniformly mixing the nine powdery raw materials according to the mass ratio of 1:5, adding warm water with the temperature of 25 ℃ for soaking overnight (12 h), adding hot water with the temperature of 78 ℃ for the next day, stirring while heating the water, fully leaching for 2h after the water temperature is 45 ℃ to obtain filtrate and filter residues, and drying the filtrate at the temperature of 60 ℃ for 120min to obtain a product A;
s4, adding distilled water into the filter residue obtained in the step S3, uniformly mixing, placing the filter residue and the distilled water in a mass ratio of 1:2 in a multi-frequency power ultrasonic microwave synergistic combination workstation, setting the ultrasonic power to be 150w, the microwave power to be 180w, the temperature to be 60 ℃, extracting for 20min, and drying for 240min at the temperature of 62 ℃ to obtain a product B;
s5, mixing the product A obtained in the step S3 with the product B obtained in the step S4 to obtain a crude product of the activator; the crude activator and snailase were then mixed according to 15:2, mixing uniformly to obtain the immune activator.
Example 3
The livestock and poultry natural immune activator comprises an activator crude product and snailase, wherein the activator crude product is prepared from the following components in parts by weight: 15 parts of aloe skin, 8 parts of dried lotus root residues, 9 parts of pine nut shells, 20 parts of pteris longifolia, 8 parts of hemerocallis, 8 parts of red hair algae, 12 parts of termitomyces albuminosus, 9 parts of lentinus edodes and 6 parts of elm; the mass ratio of the snailase to the crude activator is 2:15.
The preparation method of the livestock and poultry natural immune activator comprises the following steps:
s1, respectively screening out impurities in nine components in the crude product of the activating agent by adopting an intelligent double-layer blast type winnowing machine, respectively cleaning, and then drying at a low temperature of 57 ℃ for 15 hours to obtain each component after drying;
s2, sequentially using a shearing pulverizer to obtain the dried components in S1 in a ratio of 3×10 3 Pre-crushing for 20s at the rotating speed of r/min, and then performing secondary crushing by using a double-hammer horizontal rail type crusher with the aperture of 7.0mm to obtain nine powdery raw materials;
s3, weighing nine powdery raw materials obtained in the step S2 according to the weight portions respectively, and uniformly mixing the nine powdery raw materials according to the mass ratio of 1:5, adding warm water with the temperature of 25 ℃ for soaking overnight (13 h), adding hot water with the temperature of 78 ℃ for the next day, stirring while heating the water, fully leaching for 2h after the water temperature is 45 ℃ to obtain filtrate and filter residues, and drying the filtrate at the temperature of 70 ℃ for 120min to obtain a product A;
s4, adding distilled water into the filter residue obtained in the step S3, uniformly mixing, placing the filter residue and the distilled water in a mass ratio of 1:2 in a multi-frequency power ultrasonic microwave synergistic combination workstation, setting the ultrasonic power to be 150w, the microwave power to be 180w, the temperature to be 60 ℃, extracting for 20min, and drying for 240min at the temperature of 70 ℃ to obtain a product B;
s5, mixing the product A obtained in the step S3 with the product B obtained in the step S4 to obtain a crude product of the activator; the crude activator and snailase were then mixed according to 15:2, mixing uniformly to obtain the immune activator.
Comparative example 1
The comparative example is a common corn-bean pulp type basic ration for livestock and poultry, and comprises 59.70% of corn, 26.80% of puffed bean pulp, 2.00% of cotton pulp, 3.00% of corn protein powder, 1.40% of stone powder, 1.10% of calcium hydrophosphate, 3.50% of rice bran oil and 2.50% of premix; no immune activator is added into the basic ration.
Comparative example 2
The livestock and poultry immune activator of the comparative example comprises a crude activator product and snailase, wherein the crude activator product comprises the following components in parts by weight: 3 parts of dry lotus root residues, 4 parts of pine nut shells, 10 parts of grape fern algae with long stems, 5 parts of hemerocallis fulva, 5 parts of red hair algae, 6 parts of termitomyces albuminosus, 4 parts of lentinus edodes and 3 parts of elm; the mass ratio of the snailase to the crude activator is 2:15.
The preparation method of the livestock and poultry immune activator of the comparative example is completely the same as that of the livestock and poultry natural immune activator in the example 1.
Comparative example 3
The livestock and poultry immune activator of the comparative example comprises a crude activator product and snailase, wherein the crude activator product comprises the following components in parts by weight: 8 parts of aloe peel, 3 parts of dried lotus root residues, 4 parts of pine nut shells, 10 parts of pteris longifolia, 5 parts of hemerocallis fulva and 5 parts of red hair algae; the mass ratio of the snailase to the crude activator is 2:15.
The preparation method of the livestock and poultry immune activator of the comparative example is completely the same as that of the livestock and poultry natural immune activator in the example 1.
Comparative example 4
The livestock and poultry immune activator of the comparative example comprises a crude activator product and snailase, wherein the crude activator product comprises the following components in parts by weight: 6 parts of termitomyces albuminosus, 4 parts of lentinus edodes and 3 parts of elm; the mass ratio of the snailase to the crude activator is 2:15.
The preparation method of the livestock and poultry immune activator of the comparative example is completely the same as that of the livestock and poultry natural immune activator in the example 1.
Test 1
1) Test method
500 healthy cobb500 chickens, 1 day old, were selected from a chicken house in mountain west city, randomly divided into 5 treatments, 10 replicates each, 10 chickens each. Fully-closed automatic standard chicken house cage culture. The general corn-soybean meal type base ration of comparative example 1 (the composition and the nutrition levels of the base ration are shown in table 3), the general corn-soybean meal type base ration of comparative example 1 + the immune activator of example 1 (addition amount 0.50%), the general corn-soybean meal type base ration of comparative example 1 + the immune activator of comparative example 2 (addition amount 0.50%), the immune activator of comparative example 1 + the immune activator of comparative example 3 (addition amount 0.50%) and the general corn-soybean meal type base ration of comparative example 1 + the immune activator of comparative example 4 (addition amount 0.50%) were fed respectively. And the test period is 42 days, the feed is freely ingested during the test period, the water is sufficiently drunk, the immunity is realized according to the conventional immunization program of the broiler chickens, and the normal feeding management is realized. At the end of the test, 3 chicken wings were collected from veins in each repetition, 1 slaughter sample was selected from each repetition, and immune-related indicators were determined. Analysis of data ANOVA one-way ANOVA analysis of variance was performed using SPSS21.0 and Duncan's multiple comparisons.
TABLE 3 composition and nutrient level of ordinary corn-soybean meal base ration (air-dried base) in comparative example 1
Note that: the premix is purchased from Shaanxi sheep agriculture technology and technology Co.Ltd; the nutrient level is calculated.
2) Test results
Table 4 growth performance of broiler chickens in example 1 and comparative examples 1-4
Note that: the same column of data P <0.01 indicates that the difference is extremely significant, P <0.05 indicates that the difference is significant, and P >0.05 indicates that the difference is not significant. The table below is the same.
Table 4 indicates that the average daily gain and survival rate of the broiler of example 1 was highest and the feed/weight ratio and average daily feed intake were lowest among all groups. Therefore, the addition of the immune activator in the basic ration of the broiler chickens can indeed promote in-vivo circulation, improve metabolism rate and promote digestion and absorption of nutrient substances, so that the growth and development of the broiler chickens and disease resistance are obviously improved; the addition of the immune activator of example 1 gave the highest survival rate and the best effect.
TABLE 5 immune organ development of broiler chickens in example 1 and comparative examples 1-4
Table 5 indicates that the thymus, spleen and bursa index of the broiler of example 1 were the highest among all groups, indicating that the immune activator of example 1 was used to promote immune organ development and improve body immunity in the case of healthy broiler.
TABLE 6 blood cell count and classification of broiler chickens in example 1 and comparative examples 1-4
Table 6 indicates that the white blood cell and lymphocyte numbers were highest, but the monocytes were lowest for example 1 among all groups. From this, it is demonstrated that the immune activator in example 1 can inhibit inflammatory reaction in the broiler chicken by changing the relative proportion of blood cells, thereby significantly affecting the immune status of the broiler chicken.
TABLE 7 serum Immunity index of broilers from example 1 and comparative examples 1-4
Table 7 indicates that the serum IgA, igG, IFN-Y, IL-2 of the broiler chickens of example 1 is highest among all groups; the serum IgM of the broiler chickens in example 1 was not significantly different from the serum IgM of the broiler chickens in comparative examples 1-4. Thus, the addition of the immune activator in example 1 can indeed achieve the purpose of recognizing and eliminating antigenic foreign matters and improving the immune level of the organism by regulating the secretion of immune factors and immunoglobulins.
Table 8 intestinal flora (belonging to the level) of broilers in example 1 and comparative examples 1-4
The intestinal flora is an indispensable component in the animal body, and has important physiological significance, including resisting pathogen invasion, stimulating the maturation of immune organs of the organism, activating immune systems, regulating metabolism of substances and the like. The intestinal flora of animals is in a relatively stable state, and the number of the bacterial phylum does not obviously fluctuate. Table 8 shows the results of high throughput sequencing of the 16SrDNA V3+V4 region of the cecal contents of broiler chickens based on the Illumina Hiseq sequencing platform. The results indicated that the relative abundance of firmicutes in the cecal content of example 1 broilers was significantly reduced and that bacteroidetes was instead increased in all groups. The application of the immune activator in daily ration of the broiler chickens has obvious regulation effect on intestinal microbiota of the broiler chickens.
Comparative example 5
The comparative example is a livestock and poultry corn-bean pulp type basic ration, and the composition is as follows: 64% of corn, 25% of soybean meal, 6.00% of wheat bran, 1.60% of soybean oil, 1.20% of stone powder, 0.80% of calcium hydrophosphate, 0.40% of salt and 1.00% of premix; no immune activator is added into the basic ration.
Test 2
1) Test method
The experiment is carried out in Wang Cunxi pig farms of Shaanxi sheep and livestock limited, 100 PIC commercial pigs (about 25kg of weight and each half of male and female pig) are selected and randomly divided into 2 treatments, namely a control group and a test group, wherein each treatment is repeated 5 times, and each repetition is repeated 10 times. The corn-soybean meal type basic ration of comparative example 5 (the composition and nutrition levels of the basic ration are shown in Table 9) and the basic ration of comparative example 5 + the immune activator of example 3 (the addition amount is 1.20%) were fed separately. The test period is 30 days, the test period is free to eat, and sufficient drinking water is provided, so that the fattening house is immunized according to the conventional immunization program of the fattening house, and the normal feeding management is realized. At the end of the test, 3 pig wing veins were collected from each repetition, and 1 pig wing vein was slaughtered and sampled from each repetition, and immune-related indexes were measured. Analysis of data ANOVA one-way ANOVA analysis of variance was performed using SPSS21.0 and Duncan's multiple comparisons.
Table 9 composition and nutrient level of corn-soybean meal base ration (air dried base) in comparative example 5
Note that: the premix is purchased from Shaanxi sheep agriculture technology and technology Co.Ltd; the nutrient level is calculated.
2) Test results
Table 10 growth performance of pigs in example 3 and comparative example 5
Note that: the same column of data P <0.01 indicates that the difference is extremely significant, P <0.05 indicates that the difference is significant, and P >0.05 indicates that the difference is not significant. The table below is the same.
Table 10 indicates that the average daily gain and survival rate of the pigs of example 3 are significantly higher than those of comparative example 5, with the ratios of feed to feed being reversed. Therefore, the immune activator in the embodiment 3 is added into the basic ration of the growing-finishing pig, so that the internal circulation can be promoted, the metabolism rate can be increased, the digestion and absorption of nutrient substances can be accelerated, the growth and development of organisms can be obviously improved, and the disease resistance can be enhanced.
TABLE 11 blood cell count and classification of pigs in example 3 and comparative example 5
Table 11 indicates that the number of leukocytes and lymphocytes in the pigs of example 3 is significantly higher than in comparative example 5, but that the number of monocytes in the pigs of example 3 is significantly reduced. This demonstrates that the immune activator of example 3 can significantly affect the immunity of the body by inhibiting inflammatory reactions in the body by changing the relative proportions of blood cells.
Table 12 serum immune index of pigs in example 3 and comparative example 5
Table 12 indicates that pig serum IgA, igG, igM, IL-6 and IFN-Y were significantly elevated in example 3 as compared to comparative example 5. It means that the addition of the immune activator in example 3 can actually achieve the regulation of immune enhancement by affecting the secretion of the relevant immunocytokine and immunoglobulin.
Normally, the intestinal barrier can prevent toxic and harmful substances in the intestinal cavity from penetrating through the intestinal mucosa to enter other tissues and organs in the body, so that the size of the mucosa permeability directly reflects the intestinal immune condition of the body. FIG. 1 reflects the results of real-time fluorescent quantitative PCR detection of jejunal mucosa sample tight junction proteins (blue in FIG. 1 is comparative example 5, orange is comparative example 3), and shows that compared with comparative example 5, the expression level of ZO-1, claudin-1 and Occludin three jejunal mucosa tight junction proteins mRNA of the pig in example 3 is significantly up-regulated, which means that after the immune activator in example 3 is added, the intestinal mucosa of the pig in example 3 is repaired, the small intestinal permeability is reduced, and the immune state of the intestinal mucosa of the organism is significantly enhanced.
In conclusion, the immune activator is rich in various nutritional ingredients such as protein, fat, mineral substances, polysaccharide (including glucomannan) and the like, has low content of harmful metal elements, and can accelerate the growth and development of animal immune organs and influence the synthesis of antibodies in vivo, proliferation and differentiation of immune cells or secretion of immune cytokines if being fed to livestock and poultry according to the proportion of 0.50-1.20%, thereby improving humoral immunity. In addition, by affecting the intestinal barrier function and regulating the intestinal microflora of animals, the immune activator can achieve effective intestinal mucosa immunity improvement effect, the autoimmune effect is further obviously enhanced, and the death number of livestock and poultry during cultivation is obviously reduced. The preparation process is reliable, simple and convenient, and has the advantages of energy saving, low consumption, little environmental pollution and wide market prospect in the whole production process.
Example 4
The natural livestock and poultry immune activator in the embodiment comprises a crude activator product and snailase, wherein the crude activator product is prepared from the following components in parts by weight: 12 parts of aloe skin, 6 parts of dried lotus root residues, 7 parts of pine nut shells, 17 parts of pteris longifolia, 7 parts of hemerocallis, 7 parts of red algae, 8 parts of termitomyces albuminosus, 7 parts of lentinus edodes and 5 parts of elm; the mass ratio of the snailase to the crude activator is 2:15.
The above description is only of the preferred embodiments of the present invention, and is not intended to limit the present invention. Any simple modification, variation and equivalent variation of the above embodiments according to the technical substance of the invention still fall within the scope of the technical solution of the invention.
Claims (8)
1. The natural livestock and poultry immune activator is characterized by comprising a crude activator product and snailase, wherein the crude activator product is prepared from the following components in parts by weight: 8-15 parts of aloe skin, 3-8 parts of dried lotus root residues, 4-9 parts of pine nut shells, 10-20 parts of pteris longifolia, 5-8 parts of hemerocallis, 5-8 parts of red algae, 6-12 parts of termitomyces albuminosus, 4-9 parts of lentinus edodes and 3-6 parts of elm; the mass ratio of the snailase to the crude activator is 2:15.
2. A method for preparing the livestock and poultry natural immune activator as claimed in claim 1, which is characterized by comprising the following steps:
s1, cleaning each component in the crude product of the activator respectively, and then drying at a low temperature to obtain each component after drying;
s2, sequentially pre-crushing the dried components obtained in the step S1, and then secondarily crushing to obtain nine powdery raw materials;
s3, respectively weighing the nine powdery raw materials obtained in the step S2, uniformly mixing, adding warm water for soaking overnight, adding hot water, heating the water while stirring, leaching for 2 hours after the water temperature is 45 ℃, filtering to obtain filtrate and filter residues, and drying the filtrate to obtain a product A;
s4, adding distilled water into the filter residue obtained in the S3, uniformly mixing, then placing the mixture in a multi-frequency power ultrasonic microwave synergistic combination workstation for extraction, and drying to obtain a product B;
s5, mixing the product A obtained in the step S3 with the product B obtained in the step S4 to obtain a crude product of the activator; and then uniformly mixing the crude activator product with snailase to prepare the immune activator.
3. The method for preparing the natural livestock and poultry immune activator according to claim 2, wherein the low-temperature drying temperature in S1 is 45-57 ℃ and the time is 15h.
4. The method for preparing a natural immunostimulant for livestock and poultry according to claim 2, wherein the pre-pulverization in S2 is performed by using a shearing pulverizer at a rotation speed of 3×10 3 r/min, the time is 10-20s; the secondary crushing uses a double-hammer horizontal rail type crusher with the aperture of 7.0 mm.
5. The method for preparing the natural livestock and poultry immune activator according to claim 2, wherein the mass ratio of the total mass of the nine powdery raw materials to the warm water in S3 is 1:5; the volume ratio of the hot water to the warm water is 5:1; the temperature of the warm water is 25 ℃, and the temperature of the hot water is 78 ℃.
6. The method for preparing a natural livestock and poultry immune activator according to claim 2, wherein the drying temperature in S3 and S4 is 55-70 ℃, the drying time in S3 is 120min, and the drying time in S4 is 240min.
7. The method for preparing the natural livestock and poultry immune activator according to claim 2, wherein the mass ratio of the filter residue to the distilled water in the S4 is 1:2; the ultrasonic power of the multi-frequency variable power ultrasonic wave and microwave cooperative combination workstation is 150w, the microwave power is 180w, the temperature is 60 ℃, and the extraction time is 20min.
8. The use of the natural livestock and poultry immune activator as claimed in claim 1, wherein the immune activator is added into the basic ration of livestock and poultry in a mass ratio of 0.50-1.20%.
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