A kind of laying hen feed and application thereof

Abstract

This invention discloses a novel layer hen feed and its application, relating to the field of feed. The novel layer hen feed additive, by weight, comprises the following raw material components: Astragalus membranaceus 15-35 parts, Marigold 10-35 parts, Gardenia jasminoides 10-40 parts, Terminalia chebula 5-15 parts, Citrus reticulata 5-15 parts, Alfalfa 5-20 parts, and Cuscuta chinensis 5-15 parts. Adding this novel layer hen feed additive to the basal diet of layer hen can increase egg weight, egg shape index, yolk color, and eggshell thickness, inhibit inflammation, and significantly improve the diversity of intestinal flora and the abundance of probiotics in fed poultry. This novel layer hen feed additive can improve the production performance of layer hens, enhance egg quality, and provide an antibiotic-free method for raising layer hens, producing greener, safer eggs with controllable internal and external quality, meeting the growing demand for safe eggs.

Description

Technical Field

[0001] The invention relates to the field of feed, in particular to a novel laying hen feed and application thereof. Background Art

[0002] The feed industry has been using antibiotics to fight poultry and livestock diseases for a century. However, the abuse of antibiotics in feed has led to antibiotic residues in livestock and poultry products and the production of drug-resistant microorganisms. For the poultry farming industry, the long-term use of antibiotics will also lead to a decrease in the immunity of laying hens, increase the probability of epidemics, and seriously endanger public health and safety. In addition, there are various stresses in the production environment of laying hens. If the free radicals in the body cannot be discharged in time, it will cause metabolic disorders in the body, and it is easy to have problems with decreased egg production quality caused by oxidation and inflammation. With the continuous improvement of social living standards and consumption capacity, people's demand for livestock and poultry products is increasing. How to avoid the side effects of antibiotics while maintaining the output of poultry and livestock products has become an urgent problem to be solved in the current farming industry.

[0003] In recent years, the feed industry has conducted more and more research on low-antibiotic, antibiotic-free, and antibiotic-alternative feeds, and the search for effective antibiotic-alternative products has become a research hotspot in the industry. Current antibiotic-alternative products mainly include: Chinese herbal preparations, probiotic preparations, enzyme preparations, antimicrobial peptides, and organic trace elements. Compared with antibiotics, antibiotic-alternative products are green feed additives with no toxic side effects, no drug residues, and no environmental pollution. However, the cost of ordinary antibiotic-alternative feed products has increased year by year, and their effects on improving egg quality are also mixed. Summary of the invention

[0004] The invention provides a novel laying hen feed and application thereof, which can improve the production performance of laying hens, improve egg quality and reduce production costs without adding antibiotics.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] One of the technical solutions of the present invention is a novel laying hen feed additive (BHCHECK-2, the name of the novel laying hen feed additive of the present invention, the BHCHECK-2 described below is equivalent to the novel laying hen feed additive of the present invention), which comprises the following raw material components by weight: 15 to 35 parts of astragalus, 10 to 35 parts of marigold, 10 to 40 parts of gardenia, 5 to 15 parts of terminalia chebula, 5 to 15 parts of tangerine peel, 5 to 20 parts of lucerne, and 5 to 15 parts of dodder seed.

[0007] The main components of Astragalus are astragalus polysaccharides and astragalosides. Astragalus polysaccharides have the effects of stimulating the development of immune organs, enhancing immunity and anti-oxidation, and regulating intestinal flora.

[0008] Marigold is rich in carotenoids, among which lutein and lutein esters account for a large proportion. Lutein has excellent ability to remove superoxide anions and peroxides, and is a very good natural antioxidant. Poultry cannot synthesize lutein by itself, so adding lutein to the diet can improve the antioxidant capacity of poultry. Marigold is also rich in natural pigments. Adding it to chicken feed can make laying hens produce deep orange yolks and inhibit yolk lipid oxidation, and make broiler skin golden. However, if it is added in excess, the egg color will be too dark, close to the color of artificial eggs.

[0009] Gardenia is also a source of high-quality natural yellow pigments. It also has anti-inflammatory effects, improves the production performance of laying hens, and improves egg quality. The ingredients of gardenia contain iridoterpenes such as gardenia glycosides, which have excellent antioxidant properties and can effectively resist the attack of free radicals on the concentrated protein secreted by eggs. It also contains natural pigments such as gardenia yellow pigment, which can make the egg yolk dark yellow and increase the egg production rate. Compared with traditional artificial synthetic pigments, it not only does not produce corresponding side effects, but also has higher quality and better health effects. Eggs produced by adding a certain amount of gardenia can also reduce their cholesterol and blood sugar levels, maximizing the quality of eggs. However, if used excessively, the gardenia glycosides in it may have certain side effects on the chicken liver.

[0010] Tangerine peel mainly contains volatile oils, flavonoids and other chemical components. It has the functions of bidirectional regulation of gastrointestinal motility, anti-inflammatory, antibacterial, antioxidant and regulation of intestinal flora. It is a natural antioxidant. It can also improve the production performance of laying hens and improve egg quality.

[0011] Alfalfa is an important forage crop with very high nutritional value and can improve and maintain the function of the digestive system.

[0012] The combination of the above ingredients synergistically promotes the growth and organ development of laying hens and increases egg production.

[0013] The second technical solution of the present invention is a method for preparing the above-mentioned novel laying hen feed additive, comprising the following steps:

[0014] The raw materials are crushed and extracted to obtain an extract; the extract is concentrated under reduced pressure and dried to obtain a dry paste; the dry paste is crushed and then added with wheat bran to granulate; thus, the novel laying hen feed additive is obtained.

[0015] The pulverizing specifically comprises pulverizing the raw material into coarse powder with a particle size of 80 to 300 meshes.

[0016] The extraction is specifically as follows: the coarse powder is separately or mixed and then soaked in water 2 to 8 times the mass of the coarse powder for extraction once to obtain an extract 1 and a residue; the residue is separately or mixed and then soaked in 95% ethanol 2 to 6 times the mass of the residue for extraction 1 to 3 times, and the ethanol is recovered under reduced pressure to obtain an extract 2; the extract 1 and the extract 2 are mixed to obtain the extract.

[0017] The water soaking extraction temperature is 60-70°C, the time is 2h, and the pressure is normal pressure;

[0018] The 95% ethanol immersion extraction is performed at a temperature of 50-55°C, a time of 2 hours, and a pressure of normal pressure;

[0019] The temperature of the reduced pressure recovery is 40-55°C and the pressure is -0.1MPa.

[0020] Using water and ethanol for two extractions can extract the effective ingredients in the coarse powder into the extract to the greatest extent. Too much water or ethanol will lead to a long subsequent concentration and drying time, large resource consumption, and high cost; too little water or ethanol will cause insufficient extraction due to the limited solubility of the effective ingredients, resulting in waste. Extraction with a mass volume ratio of 2 to 8 times can extract 20% to 50% of the effective ingredients in the raw material components, which is the best ratio after multiple tests.

[0021] The drying is specifically performed by hot air drying at 60-80° C. for 2-4 hours.

[0022] Appropriate drying temperature can improve drying efficiency without destroying the effective ingredients. If the temperature is too low, the drying time required will be long, resulting in a waste of resources. If the drying temperature is too high, the effective ingredients will be destroyed, resulting in a decrease in the activity of the substances that produce the effect, thereby reducing the effect of additives in improving egg quality.

[0023] The added amount of the wheat bran is 1 to 10 times the mass of the dry paste.

[0024] Wheat bran is a byproduct of semolina production. It is rich in nutritional value and contains a large amount of dietary fiber and non-starch polysaccharides that can regulate intestinal health. Wheat bran can improve the production performance of laying hens, improve egg quality, and increase the utilization rate of total dry matter and total phosphorus in the diet. If the addition amount is too low, the effect is not significant, and if the addition amount is too high, it may have a negative impact on the feeding effect, such as reducing the apparent total intestinal digestibility of gross energy and nutrients.

[0025] The third technical solution of the present invention is a novel laying hen feed, comprising the novel laying hen feed additive mentioned above.

[0026] The addition amount of the novel laying hen feed additive in the novel laying hen feed is 1 to 3 kg / t.

[0027] If the amount of BHCHECK-2 added is too small, the effect will not be significant, and if it is added too much, it may cause toxic side effects.

[0028] A fourth technical solution of the present invention is a method for improving egg quality, wherein the novel laying hen feed is used as the daily diet for laying hens at the peak of egg production.

[0029] The reason for adding it during the peak egg-laying period is that conducting experiments during the peak egg-laying period can better prove the effectiveness of the formula, and the products produced will be more popular in the market.

[0030] The present invention discloses the following technical effects:

[0031] (1) The novel laying hen feed additive of the present invention can be added to the basic diet of laying hens to improve egg weight, egg shape index, yolk color and eggshell thickness.

[0032] (2) The novel laying hen feed additive of the present invention can reduce the content of pro-inflammatory factors in the body of fed poultry, thereby inhibiting the occurrence of inflammation; and can significantly increase the diversity of bacterial flora and the abundance of probiotics in the small intestine of fed poultry.

[0033] (3) The novel laying hen feed additive of the present invention can improve the production performance of laying hens and enhance the egg quality without adding antibiotics, thereby providing an antibiotic-free laying hen breeding method, which can produce greener and safer eggs and meet people's growing demand for safe eggs. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiment drawings of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0035] Figure 1 The effects of the new laying hen feed prepared in Examples 4-6 and Comparative Example 1 on the antioxidant indexes of laying hen livers, Figures A and B are comparison diagrams of liver MDA content levels, Figure C is a comparison diagram of liver SOD activity, and Figure D is a comparison diagram of egg yolk MDA content levels, * indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001;

[0036] Figure 2 Effects of laying hen feeds prepared in Examples 4-6 and Comparative Example 1 on the intestinal morphology of laying hens;

[0037] Figure 3 Effects of laying hen feeds prepared in Examples 4-6 and Comparative Example 1 on intestinal histology of laying hens;

[0038] Figure 4 Effects of laying hen feed prepared in Examples 4-6 and Comparative Example 1 on the expression of intestinal related genes;

[0039] Figure 5 Effects of laying hen feeds prepared in Examples 4-6 and Comparative Example 1 on the diversity of small intestinal microorganisms of laying hens;

[0040] Figure 6 Effects of laying hen feeds prepared in Examples 4-6 and Comparative Example 1 on the microbial composition of the small intestine of laying hens;

[0041] Figure 7 The effects of the laying hen feed prepared in Example 5 and Comparative Example 1 on the serum immune indicators and inflammatory factors in the body fluids of laying hens. DETAILED DESCRIPTION

[0042] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0043] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0044] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0045] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0046] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0047] Unless otherwise specified, the "parts" described in the present invention are all based on mass parts.

[0048] The "room temperature" described in the present invention is within the range of 20 to 30°C unless otherwise specified.

[0049] The present invention adds a novel laying hen feed additive (hereinafter referred to as BHCHECK-2) to a basic diet during the peak egg-laying period to verify the effect.

[0050] The basic diet formula of the present invention is a corn-soybean meal type, which is prepared according to the nutritional requirements of laying hens in China's "Chicken Feeding Standard (NY / T33-2004)". The basic diet composition formula and nutritional level are shown in Table 1:

[0051] Table 1 Basic diet composition and nutritional level (air-dry basis, %)

[0052]

[0053]

[0054] a The premix provides per kg of feed: Fe 60mg, Cu 11mg, I 0.4mg, Zn 70mg, Mn 115mg, Se0.30mg; VA, 6000IU; VD3, 2500IU; VE, 25.0mg;

[0055] VK3, 2.25mg; VB1, 1.8mg; VB2, 7.0mg; VB6, 4.0mg; VB 12 , 0.20mg; pantothenic acid, 12.0mg; niacin, 35.0mg; biotin, 0.14mg; folic acid, 0.8mg.

[0056] b Nutrient levels are calculated values.

[0057] The laying hens used in the present invention were purchased from a laying hen breeding base in Taoyuan County, Hunan Province. 55-week-old Beijing powder laying hens with similar egg production rate and consistent health conditions were selected and divided into a blank control group (control), a low-dose group (NA-L), a medium-dose group (NA-M), and a high-dose group (NA-H) using a single-factor completely randomized design.

[0058] Example 1

[0059] Weigh 30 parts of Astragalus, 25 parts of Marigold, 25 parts of Gardenia, 10 parts of Terminalia chebula, 10 parts of Tangerine Peel, 15 parts of Medicago truncatula, and 10 parts of Cuscuta australis;

[0060] The raw materials were crushed into coarse powder with a particle size of 100 mesh, mixed, and soaked in water 6 times the mass of the raw materials at 60° C. for 2 hours, filtered to obtain extract 1 and residue; then, the residue was soaked in 95% ethanol 5 times the mass of the residue at 50° C. for 2 hours, and the same amount of ethanol was used for extraction (a total of 2 ethanol extractions), the ethanol was mixed and recovered under reduced pressure to obtain extract 2, and the extract 1 and the extract 2 were mixed to obtain an extract;

[0061] The extract was concentrated under reduced pressure at 50°C and -0.1MPa; then dried at 70°C for 3h to obtain a dry paste;

[0062] The dry paste was crushed and granulated with wheat bran in an amount 8 times the mass of the dry paste to obtain BHCHECK-2①.

[0063] Example 2

[0064] Weigh 35 parts of Astragalus, 35 parts of Tagetes, 40 parts of Gardenia, 15 parts of Terminalia chebula, 15 parts of Tangerine Peel, 20 parts of Medicago, and 15 parts of Cuscuta;

[0065] The raw materials are crushed into coarse powders with a particle size of 80 mesh, and soaked in water 8 times the mass of each component at 70° C. for 2 hours, and filtered to obtain an extract 1 and a residue of each component; then soaked in 95% ethanol 6 times the mass of the residue of each component at 55° C. for 2 hours, and the residue of each component is extracted twice with an equal amount of ethanol (each component is extracted with ethanol three times in total), the ethanol used for each extraction of each component is mixed and recovered under reduced pressure to obtain an extract 2 of each component, the extract 1 of all components is mixed to obtain an extract 1, the extract 2 of all components is mixed to obtain an extract 2, and the extract 1 and the extract 2 are mixed to obtain an extract;

[0066] The extract was concentrated under reduced pressure at 55°C and -0.1MPa; then dried at 80°C for 2.5h to obtain a dry paste;

[0067] The dry paste was crushed and granulated with wheat bran 10 times the mass of the dry paste to obtain BHCHECK-2②.

[0068] Example 3

[0069] Weigh 15 parts of Astragalus, 10 parts of Tagetes, 10 parts of Gardenia, 5 parts of Terminalia chebula, 5 parts of Tangerine Peel, 5 parts of Medicago truncatula, and 5 parts of Cuscuta australis;

[0070] The raw materials were crushed into coarse powder with a particle size of 120 mesh, mixed, and soaked in water 7 times the mass of the raw materials at 70°C for 2 hours, filtered to obtain extract 1 and residue; then, the residue was soaked in 95% ethanol 6 times the mass of the residue at 50°C for 2.5 hours, the ethanol was recovered under reduced pressure to obtain extract 2, and the extract 1 and the extract 2 were mixed to obtain an extract;

[0071] The extract was concentrated under reduced pressure at 40°C and -0.1MPa; then dried at 60°C for 3h to obtain a dry paste;

[0072] The dry paste was crushed and granulated with wheat bran in an amount 1 times the mass of the dry paste to obtain BHCHECK-2③.

[0073] Example 4

[0074] During the peak egg-laying period, 1kg / t of BHCHECK-2① was added to the basal diet of Jingfen laying hens as a low-dose group (NA-L).

[0075] Example 5

[0076] The same as Example 4, the only difference is that 2 kg / t of BHCHECK-2① is added to the basal diet as the medium dose group (NA-M).

[0077] Example 6

[0078] The same as Example 4, the only difference is that 3 kg / t of BHCHECK-2① was added to the basal diet as a high-dose group (NA-H).

[0079] Comparative Example 1

[0080] The same as Example 4, except that BHCHECK-2① was not added, serving as a blank control group (control).

[0081] The feeds prepared according to Examples 4-6 and Comparative Example 1 were fed to laying hens during the peak egg-laying period, with 72 laying hens in each group. The test period was 63 days, and the egg production rate, egg weight, feed intake, feed-egg ratio, mortality rate, etc. of the laying hens were recorded during the entire period.

[0082] Effect verification example 1: Effect of BHCHECK-2① on egg quality

[0083] Egg samples were collected one day before the end of the experiment (day 62), and 8 eggs were randomly selected from each group for egg quality testing. The testing method was as follows:

[0084] Egg weight: measured using Egg Analyzer, accurate to 0.01g;

[0085] Egg shape index: Use an electronic digital display vernier caliper to measure the horizontal and vertical diameters of each egg, and then calculate the egg shape index according to formula 1:

[0086] Egg-shaped index = horizontal diameter / vertical diameter × 100% Formula 1;

[0087] Eggshell strength: Place the intact eggs on an eggshell strength tester for testing.

[0088] Yolk color: Crack the eggs and place them in the Egg Analyzer for testing.

[0089] Haugh units and albumen height: Crack the eggs and place them in the Egg Analyzer for testing.

[0090] Eggshell thickness: Use an eggshell thickness gauge to measure the eggshell thickness at the sharp end, blunt end and middle part of the egg, and take the average value as the eggshell thickness.

[0091] Yolk index: Use an electronic digital display vernier caliper to measure the yolk height and yolk diameter, and calculate the yolk index according to Formula 2:

[0092] Yolk index = yolk height / yolk diameter Formula 2;

[0093] Yolk ratio: remove the protein and ligaments on the yolk surface, weigh and record the yolk weight and the whole egg weight on an analytical balance, and calculate the yolk ratio according to formula 3:

[0094] Yolk ratio = yolk weight / whole egg weight formula 3;

[0095] The above test results are shown in Table 3:

[0096] Table 3 Effect of BHCHECK-2① on laying hen egg quality

[0097]

[0098]

[0099] Egg weight: Compared with the blank control group, the NA-M and NA-H groups had increased egg weight;

[0100] Egg shape index: Egg shape does not affect edible value, but it is related to breeding value and hatching rate of eggs, and is very important for packaging and transportation. The suitable egg shape index has low broken and cracked eggs. When the egg shape index is higher than 80% or lower than 70%, the eggshell breakage rate increases significantly. Compared with the blank control group, the egg shape index of the NA-L, NA-M and NA-H groups was significantly improved and kept within a reasonable range (P<0.01);

[0101] Eggshell thickness: Eggshell thickness is the main factor that constitutes the compressive strength of the eggshell and is an important quality and economic indicator of eggs. Eggshell thickness has a great influence on the breakage rate. The breakage rate is 2% to 5% when the shell thickness is above 0.30mm, and the breakage rate may be as high as 10% when the shell thickness is 0.27mm to 0.29mm. When the shell thickness is less than 0.25mm, the breakage rate will be as high as 85%. Compared with the blank control group, the NA-M and NA-H groups can significantly increase the eggshell thickness (0.30mm to 0.32mm);

[0102] Yolk color: Yolk color affects consumers' choice of eggs. Consumers usually prefer dark yolks because dark yolks look more natural. In this case, the yolk color increases linearly with the increase of BHCHECK-2① addition level.

[0103] In conclusion, the addition of different doses of BHCHECK-2① can improve egg quality to varying degrees, and all egg quality indicators of the NA-M and NA-H groups were greatly improved.

[0104] Effect verification example 2: Effect of BHCHECK-2① on serum biochemical indexes of eggs

[0105] The indicators of immunoglobulin A (IgA) and immunoglobulin M (IgM) in the serum of each group of Jingfen laying hens were determined using Kehua automatic biochemical analyzer (serum biochemical related kits were purchased from Kehua Biology). The results are shown in Table 4:

[0106] Table 4 Effects of BHCHECK-2① on serum biochemical parameters of laying hens

[0107]

[0108] As shown in Table 4, compared with the blank control group, all groups with NA added had the effect of increasing the content of IgM and IgA. Immunoglobulin is a protein with antibody activity. Chicken immunoglobulin can increase the activity of lysozyme and β-lysin in the body. Therefore, increasing the content of immunoglobulin in laying hens can improve their natural resistance, thereby improving the survival rate and the chicken's antibacterial and antiviral immunity.

[0109] Effect verification example 3: Effect of BHCHECK-2① on antioxidant indexes of laying hens

[0110] One day before the end of the experiment (day 62), 8 eggs were randomly selected from each group, the yolks of the selected eggs were separated from the egg whites, and stored at -30°C. The malondialdehyde (MDA) content level in the yolk, the malondialdehyde (MDA) content level in the liver, and the superoxide dismutase (SOD) activity were determined according to the kit instructions, using the relevant kits of Beijing Box Biotechnology Co., Ltd.

[0111] The results are as follows Figure 1 As shown, Figure 1In the figure, A and B are the comparison charts of liver MDA content level, C is the comparison chart of liver SOD activity, and D is the comparison chart of egg yolk MDA content level. * indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001. Compared with the blank control group, the NA-L group and the NA-M group can significantly increase the liver MDA content level, the NA-L group can significantly increase the liver SOD activity (P<0.05), and the NA-H group can significantly increase the egg yolk MDA content level (P<0.05). The above results show that the addition of BHCHECK-2① significantly increases the content level of egg yolk MDA and liver MDA, and the activity of liver SOD, and has the effect of improving the body's oxidative performance.

[0112] Effect verification example 4: Effect of BHCHECK-2① on intestinal tissue morphology

[0113] After the intestinal samples of each group of Jingfen laying hens were fixed with 4% paraformaldehyde and made into paraffin sections, the villus height and crypt depth of the duodenum, ileum and jejunum were measured using image analysis software, and the villus height / crypt depth (V / C) was calculated. The results are shown in the figure. Figure 2 , Figure 3 shown.

[0114] according to Figure 2 It can be seen that the villi of the small intestine of the three groups with NA added to the duodenum were slender, neatly arranged, and had clear brush borders. The brush border of the NA-L group was the clearest, with more and evenly distributed goblet cells and moderate crypt depth. The goblet cells of the blank control group, NA-M, and NA-H groups were less than those of the NA-L group, but they were also improved. The villi microstructure of the blank control group of the ileum was normal, with moderate villus height and width, and the columnar epithelial cells of the villi were closely arranged; compared with the blank group, the villi of the NA-L ileum were significantly shortened, the width was narrowed, and the number of columnar epithelial cells decreased. The villi of the small intestine of the other groups were slender, and all improved. In the jejunal blank control group, the morphological structure of the small intestinal villi epithelial cells was relatively complete and clear, but the number of goblet cells was small, and the crypt morphological structure was complete and the number was large. Compared with the blank group, the jejunal small intestinal villi in the NA-L and NA-M groups were more slender, with more goblet cells, and the crypt morphological structure was complete and the number was large. In the NA-H group, the small intestinal villi morphology was unclear, the goblet cells were small, there were a few crypts but the structure was unclear.

[0115] Compared with the blank control group, the villus length of the three groups supplemented with NA in the duodenum increased, and the crypt depth of the NA-L and NA-H groups decreased. Figure 3 It can be seen that the ratio of villus length to crypt depth (V / C) in the NA-L group was significant.

[0116] Intestinal tissue morphology has a great influence on the feeding and digestion of laying hens and the absorption of intestinal nutrients. Generally speaking, the intestinal tissue morphology is good if the villi are slender, the goblet cells are numerous, and the crypts are numerous and shallow. The larger the ratio of villus length to crypt depth (V / C), the stronger the nutrient absorption capacity. The above results show that the addition of BHCHECK-2① in the high-dose group can significantly improve the intestinal tissue morphology of laying hens, thereby promoting the nutrient absorption of laying hens, which is beneficial to the production of laying hens and improving the egg quality.

[0117] Effect verification example 5: Effect of BHCHECK-2① on intestinal related gene expression

[0118] Detection of inflammatory factors in the cecum: interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 10 (IL-10), and tumor necrosis factor-α;

[0119] The results are as follows Figure 4 As shown, compared with the blank control group, the contents of pro-inflammatory factors IL-6 and IL-1β in the NA-L group were significantly reduced, the contents of pro-inflammatory factors IL-6, IL-1β, and TNF-α in the NA-M group were significantly reduced, and the content of TNF-α in the NA-H group was significantly reduced; the content of anti-inflammatory factor IL-10 in NA-M and NA-H increased. This shows that adding BHCHECK-2① to feed can inhibit inflammation to varying degrees, thereby improving the production performance of laying hens.

[0120] Effect Verification Example 6: Intestinal Microbial Analysis

[0121] The microbial composition of the contents of the foregut (duodenum, jejunum, and ileum) and hindgut (cecum and colorectum) was analyzed using 16S rDNA high-throughput sequencing technology.

[0122] according to The total DNA of the microbial community was extracted according to the instructions of the soil DNA kit (Omega Bio-tek, Norcross, GA, US), and the DNA concentration and purity were determined using NanoDrop2000. The V3-V4 variable region of the 16S rRNA gene was amplified by PCR using 338F (5'-ACTCCTACGGGAGGCAGCAG-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3'), and the recovered products were purified using the AxyPrep DNAGel Extraction Kit (Axygen Biosciences, Union City, CA, USA), and the products were purified using Quantus Biosciences. TMFluorometer (Promega, USA) was used to detect and quantify the recovered products. The library was constructed using NEXTflexTM Rapid DNA-Seq Kit (Bioo Scientific, USA). Sequencing was performed using Illumina's Miseq PE300 / NovaSeq PE250 platform (Shanghai Meiji Biopharmaceutical Technology Co., Ltd.).

[0123] The original sequencing sequence was quality controlled using fastp (https: / / github.com / OpenGene / fastp, version 0.20.0) software, spliced ​​using Flash (http: / / www.cbcb.umd.edu / software / flash, version 1.2.7) software, and OTU clustered and chimeras were removed based on 97% similarity using uParse software (http: / / drive5.com / uparse / , version 7.1). Each sequence was annotated with species classification using RDP classifier (http: / / rdp.cme.msu.edu / , version 2.2), and compared with the Silva 16S rRNA database (version 138), with the alignment threshold set at 70%.

[0124] The results are as follows Figure 5 , Figure 6 As shown in the figure, the diversity of small intestinal microorganisms in the low-dose group (SJSLF), medium-dose group (SJSMF), high-dose group (SJSHF) and blank control group (CGF) of BHCHECK-2① was compared. The results showed that the medium-dose group (SJSMF) of BHCHECK-2① could significantly improve the Chao1, Simpson, Shanon and other indexes of microorganisms in the small intestine of laying hens, indicating that it can significantly improve the diversity of small intestinal microorganisms; the composition of small intestinal microorganisms (SJSLF, SJSMF, SJSHF) in the low, medium and high-dose groups of BHCHECK-2① was analyzed, and it was found that medium and high doses of BHCHECK-2① could significantly increase the abundance of probiotics (Bifidobacterium) in the small intestine.

[0125] Effect verification example 7: Effect of BHCHECK-2① on serum immune indexes and inflammatory factors

[0126] Gene expression was determined by qPCR, and serum immunoglobulin A (IgA), immunoglobulin M (IgM), and proinflammatory factor IL-2 were determined using Nanjing Jiancheng ELISA kit.

[0127] The results are as follows Figure 7 As shown in the figure, compared with the control group, the IgA and IgM levels in the NA-M group increased, indicating that NA-M has a promoting effect on the humoral immunity of Jingfen laying hens. Compared with the control group, the pro-inflammatory factor IL-2 content in the NA-M group decreased, indicating that BHCHECK-2① can reduce the expression of inflammatory factors in the cecum, thereby alleviating inflammation.

[0128] The experimental data of effect verification examples 1-7 were sorted using Excel 2019 software; the production performance, egg quality, organ index and serum biochemical data were analyzed by Tukey-Kramer method using SPSS23.0 statistical software, and the slice data were analyzed by one-way analysis of variance. Based on the above data, the subsequent experiments were analyzed by independent sample T test and marked as NA. The results were expressed as "mean ± standard error", P < 0.05 was significantly different, and P < 0.01 was extremely significant. The data were all plotted using GraphPad Prism 9.0 software.

[0129] The above effect verification shows that adding BHCHECK-2① to the basic diet of laying hens can increase egg weight and significantly reduce the content of proinflammatory factors IL-6, IL-1β, and TNF-α, thereby inhibiting inflammation; NA-H can significantly increase the diversity of the flora in the small intestine, and NA-M and NA-H can significantly increase the abundance of probiotics in the small intestine; NA-M has a promoting effect on the humoral immunity of Jingfen laying hens. The above experimental verification using BHCHECK-2② and BHCHECK-2③ prepared in Examples 2 and 3 of the present invention also obtained similar technical effects, which will not be repeated here.

[0130] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A laying hen feed, characterized in that: The raw materials include a laying hen feed additive; the amount of the laying hen feed additive added to the laying hen feed is 2 to 3 kg / t; The laying hen feed additive is prepared from the following raw material components by weight: 15-35 parts of astragalus, 10-35 parts of marigold, 10-40 parts of gardenia, 5-15 parts of terminalia chebula, 5-15 parts of tangerine peel, 5-20 parts of lucerne, and 5-15 parts of dodder seed; The preparation method of the laying hen feed additive comprises the following steps: The raw materials are crushed and extracted to obtain an extract; the extract is concentrated under reduced pressure and dried to obtain a dry paste; the dry paste is crushed and then added with wheat bran to granulate; thus, the laying hen feed additive is obtained; The extraction is specifically as follows: mixing the coarse powders, and then soaking and extracting them once with water 2 to 8 times the mass of the coarse powders to obtain an extract 1 and a residue; extracting the residue 1 to 3 times with 95% ethanol 2 to 6 times the mass of the residue, recovering the ethanol under reduced pressure to obtain an extract 2; and mixing the extract 1 and the extract 2 to obtain the extract.

2. The laying hen feed according to claim 1, characterized in that The pulverizing specifically comprises pulverizing the raw material into coarse powder with a particle size of 80 to 300 meshes.

3. The laying hen feed according to claim 1, characterized in that The drying is specifically performed at 60 to 80° C. for 2 to 4 hours.

4. The laying hen feed according to claim 1, characterized in that The added amount of the wheat bran is 1 to 10 times the mass of the dry paste.

5. A method for improving egg quality, characterized in that: The laying hen feed according to claim 1 is used as the daily diet for laying hens in the peak egg-laying period.

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