Water-soluble cassia twig volatile oil solution for laying fowls as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of veterinary drugs, discloses a water-soluble cassia twig volatile oil solution for laying fowls as well as a preparation method and application of the water-soluble cassia twig volatile oil solution, and particularly discloses the water-soluble cassia twig volatile oil solution which comprises the following components: cassia twig volatile oil, a preservative, an emulsifier, a co-emulsifier and an antioxidant. The water-soluble cassia twig volatile oil solution disclosed by the invention has a relatively strong in-vitro antibacterial effect, can effectively prevent and treat enteritis and salpingitis of laying fowls, improve the clinical dilute fecal symptom, reduce sand-shell eggs, soft-shell eggs and the like caused by salpingitis, improve the rate of certified eggs and prolong the egg laying peak.

Description

Technical Field

[0001] The invention belongs to the technical field of veterinary drugs, and particularly relates to a water-soluble cassia twig volatile oil solution for egg-laying poultry, and a preparation method and application thereof. Background Art

[0002] Enteritis and salpingitis are two major problems that cannot be avoided in poultry farming, especially laying hen farming. They can occur all year round, but the incidence is highest in summer. Inflammation often occurs in chicken farms with poor breeding environment and low management level. There are many pathogenic factors that lead to poultry enteritis and salpingitis. Bacterial infection is often the main factor that causes it. It is prevalent all over the world. Clinically, infections such as Escherichia coli, Salmonella, and Clostridium perfringens are the most common, which poses a great threat to the breeding industry. The disease is mostly caused by pathogenic bacteria through the mouth, and it is easy to occur when the content of microorganisms in feed or drinking water exceeds the standard.

[0003] After bacterial enteritis occurs, sick chickens usually show listlessness, stand still, droop wings, close eyes and shrink necks, and are unwilling to move. As the onset time increases, they will become thin, their combs will turn pale, their feed intake will decrease, and they will have watery or feed-like feces. In severe cases, they will have dark brown or red carrot-like feces. On the one hand, bacterial infection will reduce the body's absorption of nutrients, damage the surface of intestinal villi, and reduce the surface area of ​​​​the villi to absorb nutrients, causing nutritional deficiencies and feed waste, hindering the normal production of the flock, and seriously affecting the egg-laying performance, egg weight, and egg quality of laying hens. On the other hand, it affects the immune function of the flock's intestines, increases the probability of viral infection, and causes a large number of deaths.

[0004] Bacterial salpingitis occurs very frequently in laying hens. In mild cases, the symptoms are not obvious; in severe cases, the hens stand still with their wings drooping, like penguins; laying hens have no peak egg production, the eggshell quality is poor, the number of thin-shelled eggs, soft-shelled eggs, and sand-skinned eggs increases, and the eggshells are often stained with blood; the anus is red and swollen, and the yellow and white pus-like secretions in the oviduct are discharged from the cloaca, contaminating the feathers around the anus; when the anus is turned over for examination, there are strips, dots, and purple-red bleeding at the oviduct opening. The main hazard after inflammation of the oviduct is irregular development of follicles, which leads to a decrease in egg-laying performance and a prolongation of the egg-laying cycle. Salpingitis can also lead to a decrease in protein secretion, resulting in small eggs and light eggs, affecting the economic benefits of breeding. In addition, it can also cause secondary infections with peritonitis and respiratory symptoms.

[0005] According to big data statistics, about 76.1% of chicken flocks in the world will suffer from enteritis or salpingitis. Bacterial enteritis causes annual losses of more than 2 billion yuan in the poultry industry, and salpingitis affects laying hens, causing losses of 36.46 yuan per chicken in each production cycle. Therefore, it is of great significance to take effective prevention and control measures to deal with the increasingly serious bacterial infections in chicken flocks.

[0006] At present, drug treatment mainly relies on antibiotics. For bacterial enteritis, aminoglycosides such as gentamicin, neomycin amikacin, and apramycin are mainly used; quinolone drugs such as enrofloxacin and metronidazole are mainly used; for bacterial salpingitis, spectinomycin, lincomycin hydrochloride, doxycycline, amoxicillin, enrofloxacin, ciprofloxacin, and levofloxacin are mainly used; cephalosporins such as cefaclor are also used. For laying hens in the egg-laying period, the use of a variety of conventional antibiotics will be strictly restricted, which will directly affect production efficiency; on the other hand, excessive use of antibiotics will lead to a general increase in drug resistance. In the long run, the effectiveness of conventional treatment plans will be greatly reduced, increasing breeding costs. Therefore, it is urgent to develop alternative antibiotics with high antibacterial activity for the prevention and control of enteritis / salpingitis.

[0007] In recent years, plant essential oils have attracted extensive attention due to their high antibacterial activity. They are active substances extracted from plant roots, stems, leaves, flowers and fruits through steam distillation, solvent extraction and supercritical CO2 fluid extraction. They have a certain aromatic smell, and representative components include thymol, carvacrol, cinnamaldehyde, etc. Cinnamaldehyde is the main component of volatile oil extracted from cinnamon twigs. It is used as a natural antibacterial substance with the characteristics of no residue and no drug resistance. It has a very obvious effect on the health of animal bodies and improving production performance. For example, a patent has disclosed an antibacterial growth-promoting composite additive for feed, its manufacturing method and application, the raw materials of which contain cinnamon oil and thymol 20% to 30%, have strong antibacterial and bactericidal effects on common Gram-positive bacteria and Gram-negative bacteria, have significant disease prevention and growth-promoting effects on livestock and poultry, and can effectively replace antibiotics used in breeding. There is also a patent disclosing an essential oil for inhibiting common pathogens of meat pigeons, among which cinnamon essential oil also shows strong antibacterial activity against common pathogens of poultry (Escherichia coli, Salmonella, etc.).

[0008] After analyzing the prior art, it is found that most of the patents that have been published are directly purchased cinnamon twig volatile oil raw materials. On the one hand, the source is unclear, and on the other hand, it is difficult to control the cost of use. There are few reports on the extraction and purification process of cinnamon twig volatile oil; on the other hand, after being prepared according to the essential oil formula disclosed in the patent, it is difficult to completely dissolve in water to form a clear solution for simulated clinical use, which causes great difficulties for the application of large-scale egg-laying poultry farms. In view of these problems, it is urgent to develop a cost-effective and high-yield cinnamon twig volatile oil extraction process, and emulsify the extracted cinnamon twig volatile oil to make it quickly soluble in water and realize waterline administration, which can not only control product costs, but also meet the needs of actual breeding production work, and is of great significance to promote the treatment of enteritis / salpingitis during laying period.

[0009] Therefore, this application is specially filed. Summary of the invention

[0010] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. To this end, the present invention provides a water-soluble cassia twig volatile oil solution for laying fowl and a preparation method thereof, which can be used to prevent and treat enteritis / salpingitis in laying fowl, has obvious preventive effect in clinical application, fast treatment effect, high cure rate, and no obvious side effects on production.

[0011] The purpose of the first aspect of the present invention is to provide a water-soluble cinnamon twig volatile oil solution.

[0012] The purpose of the second aspect of the present invention is to provide a method for preparing the water-soluble cinnamon twig volatile oil solution of the first aspect of the present invention.

[0013] The third aspect of the present invention aims to provide a medicine.

[0014] The purpose of the fourth aspect of the present invention is to provide the use of the water-soluble cinnamon twig volatile oil solution of the first aspect of the present invention and / or the medicine of the third aspect of the present invention.

[0015] The fifth aspect of the present invention aims to provide a product.

[0016] In order to achieve the above object, the technical solution adopted by the present invention is:

[0017] The first aspect of the present invention provides a water-soluble cinnamon twig volatile oil solution, comprising the following components: cinnamon twig volatile oil, a preservative, an emulsifier, an emulsifier aid and an antioxidant.

[0018] In some embodiments of the present invention, the water-soluble cinnamon twig volatile oil solution includes 8 to 20 parts of cinnamon twig volatile oil, 0.01 to 0.5 parts of preservatives, 20 to 37 parts of emulsifiers, 20 to 45 parts of co-emulsifiers and 0.0001 to 0.7 parts of antioxidants, calculated by mass.

[0019] In some preferred embodiments of the present invention, the water-soluble cinnamon twig volatile oil solution includes 8-15 parts of cinnamon twig volatile oil, 0.01-0.1 parts of preservatives, 20-30 parts of emulsifiers, 20-35 parts of co-emulsifiers and 0.0001-0.7 parts of antioxidants, calculated by mass.

[0020] In some embodiments of the present invention, the emulsifier includes Tween, more preferably, the emulsifier includes polyoxyethylene hydrogenated castor oil or polyoxyethylene stearate; most preferably, the emulsifier is polyoxyethylene hydrogenated castor oil.

[0021] In some embodiments of the present invention, the co-emulsifier includes at least one of polyglycerol ester, glycerol, ethylene glycol, and diethylene glycol monoethyl ether; preferably, the co-emulsifier is polyglycerol ester and diethylene glycol monoethyl ether.

[0022] In some embodiments of the present invention, the antioxidant includes at least one of butylated hydroxytoluene, sodium bisulfite, and tert-butylhydroquinone.

[0023] In some preferred embodiments of the present invention, the antioxidant includes at least two of butylated hydroxytoluene, sodium bisulfite and tert-butylhydroquinone.

[0024] In some more preferred embodiments of the present invention, the antioxidant is a combination of dibutyl hydroxybenzoate and sodium bisulfite.

[0025] In some embodiments of the present invention, the water-soluble cinnamon twig volatile oil solution includes 10-12 parts of cinnamon twig volatile oil, 10-12 parts of polyglycerol ester, 20-25 parts of polyoxyethylene hydrogenated castor oil, 11-15 parts of diethylene glycol monoethyl ether, 0.03-0.08 parts of citric acid, 0.01-0.15 parts of butylated hydroxytoluene and 0.15-0.25 parts of sodium bisulfite.

[0026] In some embodiments of the present invention, the water-soluble cinnamon twig volatile oil solution further comprises 10 to 15 parts of water.

[0027] In some embodiments of the present invention, the cinnamon twig volatile oil is prepared by the following method:

[0028] Performing compound enzyme enzymolysis on the cinnamon twig aqueous solution, and collecting the enzymolysis solution;

[0029] The enzymatic hydrolysate is extracted and concentrated using a supercritical CO2 extraction method and a solvent method or a distillation method to obtain cassia twig volatile oil.

[0030] In some embodiments of the present invention, the added amount of the complex enzyme is 2% to 4% of the mass of cinnamon twig.

[0031] In some embodiments of the present invention, the added amount of the complex enzyme is 3% of the mass of cinnamon twig.

[0032] In some embodiments of the present invention, the complex enzyme comprises cellulase, hemicellulase, pectinase and xylanase.

[0033] In some embodiments of the present invention, the enzyme activities of the cellulase, hemicellulase, pectinase and xylanase are 80,000 to 150,000 U / g.

[0034] In some embodiments of the present invention, the enzyme activities of the cellulase, hemicellulase, pectinase and xylanase are 90,000 U / g, 100,000 U / g, 90,000 U / g and 120,000 U / g, respectively.

[0035] In some embodiments of the present invention, the mass ratio of the cellulase, hemicellulase, pectinase and xylanase is 0.5-2:1:0.5-3:1-2.

[0036] In some preferred embodiments of the present invention, the mass ratio of the cellulase, hemicellulase, pectinase and xylanase is 0.5-2:1:0.5-3:1-2.

[0037] In some preferred embodiments of the present invention, the mass ratio of the cellulase, hemicellulase, pectinase and xylanase is 1-2:1:2-3:1.

[0038] In some more preferred embodiments of the present invention, the mass ratio of the cellulase, hemicellulase, pectinase and xylanase is 1:1:2:1.

[0039] In some embodiments of the present invention, the conditions for the complex enzyme enzymatic hydrolysis treatment are pH 4.5-5.5, moisture 30%-40%, temperature 45° C.-50° C., and time 0.5 h-2 h.

[0040] In some preferred embodiments of the present invention, the conditions for the complex enzyme enzymatic hydrolysis treatment are pH 4.5-5.0, moisture 35%-40%, temperature 45° C.-48° C., and time 1 h-1.5 h.

[0041] In some more preferred embodiments of the present invention, the conditions for the complex enzyme enzymatic hydrolysis treatment are pH 4.6-4.8, moisture 35%-40%, temperature 45° C.-48° C., and time 0.5 h-1.5 h.

[0042] In some embodiments of the present invention, the conditions of the supercritical CO2 extraction are extraction pressure of 25-35 MPa, extraction temperature of 40-45°C, extraction time of 0.5-3h, and CO2 flow rate of 20-25 L / h.

[0043] In some preferred embodiments of the present invention, the conditions of the supercritical CO2 extraction are extraction pressure of 25-30 MPa, extraction temperature of 40-45°C, extraction time of 1-3 h, and CO2 flow rate of 20-25 L / h.

[0044] In some more preferred embodiments of the present invention, the conditions of the supercritical CO2 extraction are extraction pressure of 28-30 MPa, extraction temperature of 41-43°C, extraction time of 1-2 h, and CO2 flow rate of 23-25 ​​L / h.

[0045] In some embodiments of the present invention, the solvent in the solvent method is ethanol.

[0046] In some embodiments of the present invention, the conditions of the solvent method are that the ethanol concentration is 50% to 70%, the amount of ethanol is 10 to 20 times the weight of the medicinal material, and the extraction time is 1 to 3 hours.

[0047] In some preferred embodiments of the present invention, the conditions of the solvent method are as follows: the ethanol concentration is 60% to 70%, the amount of ethanol is 10 to 15 times the weight of the medicinal material, and the extraction time is 1 to 2 hours.

[0048] In some more preferred embodiments of the present invention, the conditions of the solvent method are as follows: the ethanol concentration is 60% to 70%, the amount of ethanol is 10 to 12 times the weight of the medicinal material, and the extraction time is 1 to 2 hours.

[0049] In some embodiments of the present invention, the distillation method is carried out under the conditions of a distillation temperature of 88 to 100° C. and a distillation time of 3 to 5 hours.

[0050] In some embodiments of the present invention, the enzymatic hydrolyzate is extracted and concentrated using supercritical CO2 extraction and solvent method or distillation method to obtain cassia twig volatile oil.

[0051] In some embodiments of the present invention, the concentration can be carried out using conventional methods including but not limited to vacuum concentration, reduced pressure concentration, etc.

[0052] The invention can promote the extraction and collection of cassia twig volatile oil to a greater extent through composite enzyme hydrolysis combined with supercritical CO2 extraction technology.

[0053] In some embodiments of the present invention, the particle size of the water-soluble cassia twig volatile oil solution is 10 to 80 nm.

[0054] The second aspect of the present invention provides a method for preparing the water-soluble cinnamon twig volatile oil solution of the first aspect of the present invention, comprising the following steps:

[0055] Mixing an antioxidant, a preservative and an emulsifier to obtain a mixed solution A;

[0056] Mixing the mixed solution A with the emulsifier to obtain a mixed solution B;

[0057] The mixed solution B is mixed with cassia twig volatile oil and water, and then emulsified to obtain a water-soluble cassia twig volatile oil solution.

[0058] In some embodiments of the present invention, the emulsification conditions are pressure of 15000psi to 80000psi, temperature of 35°C to 45°C, and emulsification time of 10 to 70 min.

[0059] In some embodiments of the present invention, the emulsification conditions are pressure of 30,000 psi to 70,000 psi, temperature of 35° C. to 45° C., and emulsification time of 30 to 60 min.

[0060] In some embodiments of the present invention, the emulsification conditions are pressure of 55000psi to 60000psi, temperature of 35°C to 45°C, and emulsification time of 30 to 50 min.

[0061] The third aspect of the present invention provides a medicine comprising the water-soluble cinnamon twig volatile oil solution of the first aspect of the present invention.

[0062] In some embodiments of the present invention, the medicament comprises pharmaceutically acceptable excipients.

[0063] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one of fillers, disintegrants, diluents, dispersants, excipients, stabilizers, lubricants, binders, wetting agents, flavoring agents, suspending agents, solvents, sustained-release agents, emulsifiers, absorption enhancers, surfactants, preservatives, pigments, flavors and solvents.

[0064] In some embodiments of the present invention, the filler is selected from starch, sucrose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose or glucose; the binder is selected from cellulose derivatives, alginate, starch, water, dextrin, gelatin or polyvinyl pyrrolidone; the disintegrant is selected from microcrystalline cellulose, sodium hydroxymethyl starch, cross-linked polyvinyl pyrrolidone, low-substituted hydroxypropyl cellulose or cross-linked sodium hydroxymethyl cellulose; the lubricant is selected from stearic acid, polyethylene glycol, calcium carbonate, sodium bicarbonate, micro-powder silica gel, talc or magnesium stearate; the suspending agent is selected from micro-powder silica gel, beeswax, cellulose, solid polyethylene glycol: The wetting agent is selected from glycerol, polyoxyethylene stearate, hydrogenated castor oil or lecithin; the solvent is selected from liquid polyethylene glycol, isopropyl alcohol, polyoxyethylene stearate, glycerol, propylene glycol or vegetable oil, and the vegetable oil is selected from soybean oil, castor oil, peanut oil, blending oil, etc.; the surfactant is selected from sodium dodecylbenzene sulfonate, stearic acid, polyoxyethylene-polyoxypropylene copolymer, fatty acid sorbitan or polysorbate (Tween), etc.; the flavoring agent is selected from aspartame, sucralose, flavor, stevioside, acesulfame potassium, citric acid or saccharin sodium; the preservative is selected from at least one of methyl paraben or propyl paraben.

[0065] In some embodiments of the present invention, the dosage form of the drug is selected from tablets (such as ordinary tablets, double-layer tablets, multi-layer tablets, sustained-release tablets, single-chamber controlled-release tablets, double-chamber controlled-release tablets, microporous controlled-release tablets, sublingual tablets, orally rapidly disintegrating tablets, dispersible tablets, enteric-coated tablets), pills, powders, suspensions, gels, emulsions, creams, granules, nanoparticles, capsules (such as ordinary capsules, sustained-release capsules, controlled-release capsules, capsules containing micropellets or small tablets, pH-dependent capsules containing micropellets or small tablets, gastrointestinal composite capsules), suppositories, injections, sprays and injections.

[0066] The fourth aspect of the present invention provides the use of the water-soluble cinnamon twig volatile oil solution of the first aspect of the present invention and / or the drug of the third aspect of the present invention in at least one of (1) to (9):

[0067] (1) Preparing products for preventing and / or treating chicken enteritis;

[0068] (2) preparing products for preventing and / or treating chicken salpingitis;

[0069] (3) Preparation of products for preventing and / or treating diseases caused by E. coli infection;

[0070] (4) Preparation of products for preventing and / or treating diseases caused by Salmonella infection;

[0071] (5) Preparation of products for preventing and / or treating diseases caused by Clostridial infections;

[0072] (6) Preparing products for preventing and / or treating laying hens with clinical symptoms of sandy-shelled eggs, thin-shelled eggs, and dirty eggs;

[0073] (7) Prepare products that improve egg production rate, egg quality and egg quality of laying hens;

[0074] (8) Antibacterial;

[0075] (9) Preparation of antibacterial products.

[0076] In some embodiments of the present invention, the bacteria in (8) and (9) include at least one of Staphylococcus aureus, Salmonella, Escherichia coli, and Clostridium.

[0077] In some embodiments of the present invention, the chickens described in (1) and (2) include laying hens.

[0078] The fifth aspect of the present invention provides a product, comprising the water-soluble cinnamon twig volatile oil solution of the first aspect of the present invention or the medicine of the third aspect of the present invention.

[0079] In some embodiments of the present invention, the products include but are not limited to reagents, kits and drugs.

[0080] The beneficial effects of the present invention are:

[0081] The water-soluble cassia twig volatile oil solution of the present invention has a strong in vitro antibacterial effect, can effectively prevent and treat enteritis and salpingitis in laying birds, improve clinical loose stool symptoms, reduce sandy shell eggs, soft shell eggs and the like caused by salpingitis, increase the rate of genuine eggs, and prolong the peak of egg production.

[0082] The present invention performs high-pressure homogenization and emulsification treatment on cinnamon twig volatile oil, so that the cinnamon twig volatile oil has water solubility, is more convenient for waterline administration in clinical breeding, is more easily accepted by breeders, and especially plays an antibacterial and anti-inflammatory role as an alternative to antibiotics during the egg-laying period, thereby ensuring the economic benefits of egg-laying poultry breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0083] Figure 1It is a real picture of the water-soluble cassia twig volatile oil solution used for laying fowls in Examples 1 to 15 and Comparative Example 1.

[0084] Figure 2 The HPLC chromatogram of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solution of laying fowl of Examples 1 to 2.

[0085] Figure 3 The HPLC chromatogram of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solution of laying fowl of Examples 3 to 8 is shown.

[0086] Figure 4 The HPLC chromatogram of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solution of laying fowl of Examples 9 to 14.

[0087] Figure 5 The HPLC chromatograms of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solution of the laying fowl of Comparative Example 1 and Example 15 are shown.

[0088] Figure 6 The HPLC chromatograms of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solution of the laying fowl of Example 1 stored for different time periods are shown.

[0089] Figure 7 The HPLC chromatograms of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solution of the laying birds of Example 1 and Example 12 stored for different time periods.

[0090] Figure 8 The HPLC chromatograms of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solution of laying fowls of Example 12 and Example 15 stored for different time periods.

[0091] Fig. 9 The HPLC chromatograms of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solution of laying fowl in Example 15 stored for different periods of time.

[0092] Fig.10 The statistical results of weekly mortality in each experimental group are shown in the figure. ** indicates P < 0.01.

[0093] Fig.11 This is the statistical result of weekly egg production / egg production rate of each experimental group.

[0094] Fig.12 This is the statistical result of the weekly authentic egg rate of each experimental group.

[0095] Fig.13 This is the statistical result of weekly egg quality test data of each experimental group. DETAILED DESCRIPTION

[0096] The present invention is further described in detail below through specific examples.

[0097] It should be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention.

[0098] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0099] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0100] Example 1

[0101] A water-soluble cassia twig volatile oil solution for laying fowls, comprising, by weight, 12 parts of cassia twig volatile oil, 15 parts of polyglycerol ester, 25 parts of polyoxyethylene hydrogenated castor oil, 12 parts of diethylene glycol monoethyl ether, 0.05 parts of citric acid, 0.02 parts of butylated hydroxytoluene, 0.2 parts of sodium bisulfite, and 12 parts of water;

[0102] Wherein, the preparation process of cassia twig volatile oil is as follows:

[0103] (1) Wash and cut fresh cassia twigs according to traditional Chinese herbal medicine processing techniques.

[0104] (2) Complex enzyme hydrolysis: Accurately weigh the cinnamon twig, add 3.5 times the volume of water by weight of the medicinal material, adjust the pH to 4.60-4.80, and then add complex enzymes (cellulase (90,000 U / g): hemicellulase (100,000 U / g): pectinase (90,000 U / g), xylanase (120,000 U / g) mass ratio = 1:1:2:1) at a weight of 3% of the total weight of the medicinal material. After starting the enzymatic hydrolysis program, stir and hydrolyze in the enzymatic hydrolysis tank for 1.5 h. The enzymatic hydrolysis temperature is controlled at 46±1°C. After the enzymatic hydrolysis is completed, the extract A and the medicinal residue are collected by solid-liquid separation.

[0105] (3) The extract A was subjected to supercritical CO2 extraction combined with ethanol solvent extraction, with an extraction pressure of 28 MPa, an extraction temperature of 42±1°C, an extraction time of 2 h, and a CO2 flow rate of 23 L / h; the ethanol concentration was 65%, the amount of ethanol was 12 times the weight of the medicinal material, the extraction time was 1.5 h, and then low-temperature vacuum concentration was performed to obtain cinnamon twig volatile oil.

[0106] The preparation method of the water-soluble cassia twig volatile oil solution for laying fowl comprises the following steps: dissolving butylated hydroxytoluene, citric acid and sodium bisulfite in polyglycerol ester by weight to obtain solution B; adding polyoxyethylene hydrogenated castor oil and diethylene glycol monoethyl ether to solution B to obtain solution C; adding cassia twig volatile oil and water to solution C in sequence, and performing high-pressure homogenization emulsification until the obtained nanoemulsion is clear, the pressure is 60000psi, the temperature is 40℃, and the emulsification time is 45min, so as to obtain the water-soluble cassia twig volatile oil solution for laying fowl.

[0107] Example 2

[0108] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 in that the solution contains 22 parts of polyoxyethylene hydrogenated castor oil and 15 parts of diethylene glycol monoethyl ether.

[0109] Example 3

[0110] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 only in that, in the preparation process of cassia twig volatile oil, 3% of the complex enzyme is cellulase (90,000 U / g): hemicellulase (100,000 U / g): pectinase (90,000 U / g), xylanase (120,000 U / g) in a mass ratio of 2:1:3:1.

[0111] Example 4

[0112] A water-soluble cassia twig volatile oil solution for laying fowl is different from Example 1 only in that, during the preparation of the cassia twig volatile oil, when the extract A is subjected to supercritical CO2 extraction combined with ethanol solvent extraction, the extraction time is 1 hour.

[0113] Example 5

[0114] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 only in that, in the preparation method of the water-soluble cassia twig volatile oil solution for laying fowls, high-pressure homogenization and emulsification are performed until the obtained nanoemulsion becomes clear, the pressure is 55000 psi, the temperature is 40°C, and the emulsification time is 20 min.

[0115] Example 6

[0116] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 only in that, in the preparation method of the water-soluble cassia twig volatile oil solution for laying fowls, high-pressure homogenization and emulsification are performed until the obtained nanoemulsion becomes clear, the pressure is 45000 psi, the temperature is 40° C., and the emulsification time is 45 min.

[0117] Example 7

[0118] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 only in that the cassia twig medicinal material is not subjected to composite enzyme enzymolysis treatment during the preparation of the cassia twig volatile oil. The specific process is as follows:

[0119] Accurately weigh cinnamon twig medicinal material, add 3.5 times the volume of water of the medicinal material, adjust the pH to 4.60-4.80, and perform supercritical CO2 extraction and ethanol solvent extraction, with an extraction pressure of 28MPa, an extraction temperature of 42±1°C, an extraction time of 2h, and a CO2 flow rate of 23L / h; the ethanol concentration is 65%, the amount of ethanol is 12 times the weight of the medicinal material, the extraction time is 1.5h, and then low-temperature vacuum concentration is performed to obtain cinnamon twig volatile oil.

[0120] Example 8

[0121] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 only in that, in the preparation process of cassia twig volatile oil, 3% of the complex enzyme is cellulase (90,000 U / g): hemicellulase (100,000 U / g): pectinase (90,000 U / g), xylanase (120,000 U / g) in a mass ratio of 1:2:1:3.

[0122] Example 9

[0123] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 only in that, in the preparation process of cassia twig volatile oil, cassia twig medicinal materials are extracted by water distillation instead of supercritical CO2 extraction and ethanol solvent method combined extraction, and the specific process is as follows:

[0124] (1) Wash and cut fresh cassia twigs according to traditional Chinese herbal medicine processing techniques.

[0125] (2) Compound enzyme hydrolysis: The medicinal materials were weighed in sequence according to the formula, stirred and mixed evenly, 3.5 volumes of water were added to the medicinal materials, the pH was adjusted to 4.60-4.80, and then a compound enzyme (cellulase (90,000 U / g): hemicellulase (100,000 U / g): pectinase (90,000 U / g), xylanase (120,000 U / g) mass ratio = 1:1:2:1) was added to the medicinal materials at a mass ratio of 3% of the total mass of the medicinal materials. After the enzymatic hydrolysis program was started, the enzymatic hydrolysis was carried out in the enzymatic hydrolysis tank while stirring for 1.5 h. The enzymatic hydrolysis temperature was controlled at 46±1°C. After the enzymatic hydrolysis was completed, the extract A and the medicinal residue B were collected by solid-liquid separation.

[0126] (3) The extract A is subjected to water distillation at a temperature of 95±5°C for 4 h, and then concentrated in vacuo to obtain cinnamon twig volatile oil.

[0127] Example 10

[0128] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 only in that, during the preparation of the cassia twig volatile oil, when the extracting solution A is subjected to supercritical CO2 extraction combined with ethanol solvent extraction, the extraction time is 0.5h.

[0129] Embodiment 11

[0130] A water-soluble cassia twig volatile oil solution for laying fowls, which is different from Example 1 only in that polyoxyethylene hydrogenated castor oil is replaced by polyoxyethylene stearate.

[0131] Example 12

[0132] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 in that the antioxidant butylated hydroxytoluene is replaced by tert-butylhydroquinone.

[0133] Embodiment 13

[0134] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 only in that, in the preparation method of the water-soluble cassia twig volatile oil solution for laying fowls, high-pressure homogenization emulsification is performed until the obtained nanoemulsion becomes clear, and the emulsification time is 60 minutes.

[0135] Embodiment 14

[0136] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 only in that, in the preparation method of the water-soluble cassia twig volatile oil solution for laying fowls, high-pressure homogenization and emulsification are performed until the obtained nanoemulsion is clarified, and the homogenization pressure is 40000psi.

[0137] Embodiment 15

[0138] A water-soluble cassia twig volatile oil solution for laying fowls, which differs from Example 1 only in that it does not contain sodium bisulfite.

[0139] Comparative Example 1

[0140] A water-soluble cassia twig volatile oil solution for laying fowls is different from Example 1 in that the mass parts of polyoxyethylene hydrogenated castor oil are 37 and diethylene glycol monoethyl ether is not contained.

[0141] Effect Example

[0142] 1. Effect of different preparation conditions on the enhancement of water-soluble cinnamon twig volatile oil solution in laying poultry

[0143] For verifying the advantage of the effective ingredient of water-soluble cassia twig volatile oil solution of egg-laying poultry with different preparation conditions of the present invention, therefore design corresponding investigation test, carry out comprehensive evaluation. Based on embodiment 1, first fresh cassia twig is washed, cut and other pre-treatments, extraction method comprises not carrying out composite enzyme enzymolysis (embodiment 7), composite enzyme enzymolysis; Extraction method comprises water distillation extraction (embodiment 9) and supercritical CO2 extraction, emulsification condition comprises the selection of emulsifier / co-emulsifier and high pressure homogenization emulsification condition selection, adopt orthogonal test method to assess optimal preparation condition, relate to cellulase (90,000 U / g) in composite enzymolysis extraction: hemicellulase (100,000 U / g): pectinase (90,000 U / g), xylanase (120,000 U / g) proportioning=1:1:2:1, 2:1:3:1 (embodiment 3), 1:2:1:3 (embodiment 8), and the addition ratio is 3% of the medicinal material quality. The supercritical CO2 extraction time involves 0.5h (Example 10) and 2h, the emulsifier selection involves polyoxyethylene stearate (Example 11) and polyoxyethylene hydrogenated castor oil; the ratio of emulsifier: co-emulsifier involves: 25:12, 22:15 (Example 2), 37:0 (Comparative Example 1); the pressure parameters involved in the emulsification high pressure homogenization emulsification conditions are: 40000psi (Example 14), 45000psi, 55000psi, 60000psi; the time parameters are 20min, 45min, 50min, 60min (Example 13). Different preparation conditions are carried out in sequence to prepare water-soluble cassia twig volatile oil solutions for egg poultry (i.e., Examples 1 to 15 and Comparative Example 1).

[0144] 1.1 Test items

[0145] 1.1.1 Properties

[0146] Observe the properties of the oral solution (color, clarity, presence of precipitation, turbidity, floccules, etc.) and record them (this product should be a light yellow to yellowish brown clear liquid).

[0147] 1.1.2 Content determination

[0148] Determined by high performance liquid chromatography (Appendix 0512).

[0149] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel is used as filler, acetonitrile-water (35:65) is used as mobile phase, column temperature is 35°C, detection wavelength is 290nm, and the theoretical plate number calculated based on the cinnamaldehyde peak should be no less than 3000.

[0150] Preparation of reference solution: Take an appropriate amount of cinnamaldehyde reference substance, weigh it accurately, add an appropriate amount of acetonitrile to dissolve it, and dilute it with mobile phase to make a solution containing 10 μg of cinnamaldehyde per 1 mL.

[0151] Preparation of test solution: Accurately measure 1 mL of the product, place it in a 100 mL volumetric flask, add acetonitrile to dilute to the scale, shake well, and obtain.

[0152] Comparing the contents of cinnamaldehyde in the samples of Examples 1 to 15, the content calculation formula is as follows (during the calculation process, since the reference substance has a higher content (close to 100%), it is identified as 1):

[0153]

[0154] 1.1.3 In vitro antibacterial evaluation

[0155] The broth dilution method was used to determine the effects of the water-soluble cinnamon twig volatile oil solution prepared in Examples 1 to 15 on the quality control strains of Escherichia coli ATCC 25922 and the quality control strains of Salmonella CICC. 10437, Staphylococcus aureus quality control strain CICC10384, and Escherichia coli clinical strain BBF9, the minimum inhibitory concentration MIC and minimum bactericidal concentration MBC, each sample was first diluted 8 times with pure water, 100 μL of MH broth medium was added to the 2nd to 11th columns of the 96-well plate, 200 μL of MH broth medium was added to the 12th column as a control, 100 μL of 125 mg / mL water-soluble cassia twig volatile oil solution for laying fowl was added to the 1st column, and the 2-fold dilution method was used to make the final concentrations of the water-soluble cassia twig volatile oil solution for laying fowl in the 1st to 11th columns 62.5, 31.25, 15.63, 7.81, 3.90, 1.95, 0.98, 0.49, 0.24, 0.12 and 0 mg / mL, respectively, and the final concentration of the bacterial solution was 5×10 5 The 96-well plate was placed in a biochemical incubator for 24 hours. The lowest drug dilution concentration with no obvious bacterial growth was the MIC (mg / mL) of the water-soluble cinnamon twig volatile oil solution for the laying poultry. The lowest drug dilution concentration with no bacterial growth after 24 hours of drop plate culture was the MBC (mg / mL) of the water-soluble cinnamon twig volatile oil solution for the laying poultry.

[0156] 1.2 Experimental Results

[0157] 1.2.1 Trait results

[0158] The results of the properties are shown in Table 1 and Figure 1 shown.

[0159] Comparing the properties of the water-soluble cinnamon twig volatile oil solutions for laying fowls of Example 1 and Example 11, the results show that the properties of the water-soluble cinnamon twig volatile oil solution for laying fowls prepared by using polyoxyethylene hydrogenated castor oil as an emulsifier are better than those of polyoxyethylene stearate (Example 11), and the prepared water-soluble cinnamon twig volatile oil solution for laying fowls is more stable and uniform.

[0160] Comparing the properties of the water-soluble cinnamon twig volatile oil solution for laying fowl in Example 1 and Comparative Example 1, the results show that adding diethylene glycol monoethyl ether as an emulsifier in Example 1 can obtain a better emulsification effect, and the prepared water-soluble cinnamon twig oil has more stable and uniform properties.

[0161] The properties of the water-soluble cassia twig volatile oil solutions of Example 1, Example 5 and Example 13 were compared. The results showed that the time during the high-pressure homogenization emulsification process had a certain influence on the properties of the finished product. If the emulsification time was too short, the emulsification effect could not be achieved, which would affect the clarity of the finished product. If the emulsification time was too long, demulsification would occur, which would be detrimental to the dissolution of the antibacterial substances. The preferred emulsification time was 30 min to 45 min.

[0162] The properties of the water-soluble cassia twig volatile oil solutions for laying fowl in Example 1, Example 6 and Example 14 were compared. The results showed that the pressure in the high-pressure homogenization emulsification process had a certain influence on the properties of the finished product. The preferred pressure was 45000psi to 60000psi. When the pressure was lower than 45000psi, the emulsification effect weakened, resulting in demulsification of the finished product solution, which was not conducive to the dissolution of the antibacterial substances.

[0163] Table 1 Properties of Examples and Comparative Examples

[0164]

[0165] 1.2.2 Content results

[0166] The HPLC test results of the cinnamaldehyde content in the water-soluble cinnamon twig volatile oil solution of the laying birds in Examples 1 to 14 are shown in Tables 2 to 3. The HPLC spectrum of the cinnamaldehyde standard is shown in Figure 2 The HPLC spectrum of cinnamaldehyde content in the water-soluble cinnamon twig volatile oil solution of egg-laying poultry in Examples 1 to 15 and Comparative Example 1 is shown in Figure 2 to Figure 5 , taking Example 1 as a reference, the calculation formula for the improvement rate is as follows:

[0167] Improvement rate = (cinnamaldehyde content of sample - cinnamaldehyde content of Example 1) / cinnamaldehyde content of Example 1 × 100%.

[0168] Comparing the contents of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solutions of laying fowl in Example 1 and Example 7, the results showed that before supercritical CO2 extraction, the cinnamon twig medicinal material was subjected to composite enzyme enzymolysis treatment, which was better than not subjected to composite enzyme enzymolysis, and the extraction could significantly increase the content of cinnamaldehyde in the cinnamon twig volatile oil (p<0.05).

[0169] Comparing the contents of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solutions of laying fowl in Example 1, Example 3 and Example 8, the results showed that increasing the proportion of pectinase (90,000 U / g) was more conducive to the dissolution of antibacterial substances. When the preferred composite enzyme ratio was cellulase, hemicellulase, pectinase and xylanase, and the mass ratio of the four was (1-2):1:(2-3):1, the extraction could significantly increase the content of cinnamaldehyde in cinnamon twig volatile oil (p<0.05).

[0170] Comparing the contents of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solutions of laying fowl in Example 1 and Example 9, the results showed that supercritical CO2 extraction of enzymolysis product A could significantly increase the content of cinnamaldehyde in cinnamon twig volatile oil (p<0.05) compared with distillation extraction, and was more conducive to the dissolution of antibacterial substances.

[0171] The content of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solution of laying fowl in Example 1 and Example 10 was compared. The results showed that the supercritical CO2 extraction time had a certain influence on the extraction efficiency of the effective components of the cinnamon twig volatile oil, and extraction for 2 h was better than 0.5 h. Too short extraction time was not conducive to the dissolution of antibacterial substances, and extending the extraction time to 2 h could significantly improve the content of cinnamaldehyde in the cinnamon twig volatile oil (p<0.05).

[0172] Table 2 HPLC test results of the content of cinnamaldehyde in the water-soluble cinnamon twig volatile oil solution of egg-laying poultry

[0173]

[0174] Table 3 Cinnamaldehyde content determination results

[0175]

[0176]

[0177] Note: a in Table 3 represents that the cinnamaldehyde content in the sample is significantly different from that in Example 1, p<0.05.

[0178] 1.2.3 In vitro antibacterial evaluation results

[0179] The results of in vitro antibacterial evaluation are shown in Table 4. The water-soluble cinnamon twig volatile oil solutions for laying fowl in Examples 1 to 15 and Comparative Example 1 have better antibacterial activity against Staphylococcus aureus, Salmonella and Escherichia coli, which is consistent with the effective ingredient content of cinnamaldehyde, indicating that the water-soluble cinnamon twig volatile oil solutions for laying fowl in Examples 1 to 15 of the present invention have better antibacterial effect in in vitro antibacterial effect.

[0180] Table 4 Comparison of in vitro antibacterial activity of water-soluble cinnamon twig volatile oil solution for laying fowl in Examples 1 to 15 and Comparative Example 1

[0181]

[0182] Note: MIC is expressed in mg / mL.

[0183] 2. Effects of different antioxidants on the stability of active ingredients in cinnamon twig volatile oil

[0184] In order to verify the influence of different antioxidants on the stability of the effective components of cinnamon twig volatile oil, a single factor investigation test was designed to evaluate the effects of butylated hydroxytoluene, butylated hydroxytoluene + sodium bisulfite antioxidant combination, and tert-butylhydroquinone + sodium bisulfite antioxidant combination on the stability of the effective components of water-soluble cinnamon twig volatile oil solution (cinnamon twig volatile oil, with cinnamaldehyde as an indicator) for laying poultry. Taking Example 1 as an example, the antioxidant and its composition are dissolved in polyglycerol ester, and 0.05 parts of citric acid are added to obtain solution B; 25 parts of polyoxyethylene hydrogenated castor oil and 12 parts of diethylene glycol monoethyl ether are added to solution B to obtain solution C; cinnamon twig volatile oil and water are added to solution C in sequence, and high-pressure homogenization emulsification is performed until the obtained nanoemulsion is clarified, the pressure is 60000psi, the temperature is 40°C, and the emulsification time is 45min, which is the water-soluble cinnamon twig volatile oil solution. The water-soluble cassia twig volatile oil solutions for laying fowl of Example 1, Example 12 and Example 15 were prepared in sequence, and the specific groupings are shown in Table 5.

[0185] Table 5 Grouping of different antioxidants

[0186]

[0187]

[0188] 2.1 Stability test conditions

[0189] The water-soluble cinnamon twig volatile oil solution for laying poultry was placed in a stability test chamber. According to the requirements of the technical guidelines for stability research in the Compilation of Technical Guidelines for Veterinary Drug Research, the cinnamon twig volatile oil solution was subjected to accelerated and long-term tests. The product properties and cinnamaldehyde content were examined using the same detection method as "1.1" to determine the factors affecting its stability.

[0190] The water-soluble cinnamon volatile oil solution of laying fowl was placed in a comprehensive drug stability test chamber and tested for 6 months at (40±2)℃ and relative humidity (75±5)%. Samples were taken at 0, 1, 3, and 6 months to examine the properties and cinnamaldehyde content of the samples, calculate the indicated content (%) of the main components, and analyze the change trend.

[0191] The water-soluble cinnamon twig volatile oil solution was placed in a comprehensive drug stability test chamber for 6 months at (25±2)℃ and relative humidity (60±5)%. Samples were taken at 0, 1, 3, and 6 months to investigate the main component content of the samples, calculate the main component labeled content (%), and analyze the change trend.

[0192] 2.2 Experimental Results

[0193] The HPLC spectra of cinnamaldehyde content in the egg-laying poultry of Example 1, Example 12 and Example 15 after storage for different time periods with the water-soluble cinnamon twig volatile oil solution are shown in FIG. Figure 6 to Figure 9 .

[0194] The results are shown in Table 6. Comparing Example 1 with Example 15, the results show that the butylated hydroxytoluene + sodium bisulfite antioxidant composition is more helpful in improving the accelerated / long-term stability of the active ingredients of cinnamon twig volatile oil than using butylated hydroxytoluene antioxidant alone. Comparing Example 1 with Example 12, the results show that the butylated hydroxytoluene + sodium bisulfite antioxidant composition is better than the tert-butylhydroquinone + sodium bisulfite antioxidant composition in accelerating / long-term stability of the active ingredients of cinnamon twig volatile oil.

[0195] The accelerated / long-term stability of the active ingredients of the cinnamon twig volatile oil of Example 1 is better, and the selected antioxidant composition can significantly increase the content of cinnamaldehyde in the cinnamon twig volatile oil (p<0.05), indicating that the antioxidant ratio and composition within the scope of the embodiment of the present invention are more advantageous in improving the stability of the active ingredients. The accelerated / long-term stability change trend of the active ingredients of cinnamon twig volatile oil is consistent, indicating that the stability of cinnamon twig volatile oil is not sensitive to temperature.

[0196] Table 6 Stability test results

[0197]

[0198]

[0199] Note: a in Table 6 represents that the cinnamaldehyde content in the sample is significantly different from that in Example 1, p<0.05.

[0200] 3. Clinical evaluation of the treatment effect of enteritis / salpingitis in laying hens

[0201] Location: A laying hen breeding company in Nanyang City, Henan Province

[0202] Test subjects: About 119,826 laying hens at 400 days of age. Recently, the number of dirty eggs, broken eggs, and soft-shell eggs in the flock has increased, the feces are poorly formed, and autopsy shows inflammation of the intestines and bleeding spots in the fallopian tubes. The diagnosis is that there is a combination of enteritis and fallopian tube inflammation.

[0203] Experimental groups: 1) 19965 birds in the blank group (without any drug); 19978 birds and 19973 birds in the experimental groups 1 and 2 respectively (using the water-soluble cassia twig volatile oil solution for laying birds in the formula of Example 1 and Example 2); 19981 birds in the experimental group 3 (using the compound feed additive in the patent document CN115624108A: 1g carvacrol, 2g thymol and 2g eugenol); 19963 birds in the experimental group 4 (using the essential oil phytochemicals in the patent document CN115444898A: 7 parts of dandelion essential oil, 18 parts of cinnamon essential oil, 9 parts of oregano oil, 1.5 parts of forsythia essential oil, 30 parts of compound Chinese medicine extract, 45 parts of 1,2-propylene glycol, 15.4 parts of soybean lecithin, and 0.1 parts of vitamin E); 19966 birds in the experimental group 5 (antibiotic treatment group, neomycin sulfate + lincomycin).

[0204] Experimental method: The drugs of Experimental Groups 1 to 2 were administered through drinking water. According to the total daily drinking water volume, the drugs of Experimental Groups 1 to 2 were administered at a ratio of 1:6000; the feeding of Experimental Group 3 was supplemented with 0.5 mg / kg (calculated based on the essential oil content in the feed additive); the Experimental Group 4 was administered with drinking water at a ratio of 1:2000; the Experimental Group 5 was administered according to the recommended dose of antibiotics. Each group was administered continuously for 5 days, and was observed continuously for 7 days since the administration. The drinking water, feeding, feces, egg production rate, quality egg rate, eggshell quality (egg weight, eggshell thickness, eggshell strength, specific detection was carried out according to the eggshell quality evaluation method in the document "Effects of glycine and iron and their complexes on production performance and eggshell quality of laying hens in the late laying period"), mortality rate and other indicators were recorded at four time points: 1 day before administration (recorded as Day 1), 3 days after administration (recorded as Day 4), 5 days after administration (recorded as Day 6), and 2 days after administration (recorded as Day 8). The effect advantage of cassia twig volatile oil in the embodiment of the present invention in the clinical treatment of salpingitis and enteritis in laying hens was tested.

[0205] result:

[0206] (1) Statistics of water intake

[0207] The statistics of water drinking during the treatment period are shown in Table 7. There was no significant difference in water drinking among the groups (P>0.05), which proved that the drugs in each group would not affect the palatability of the chickens.

[0208] Table 7 Feed intake statistics

[0209]

[0210]

[0211] (2) Feed intake statistics

[0212] The statistics of food intake during the treatment period are shown in Table 8. There was no significant difference in food intake among the groups (P>0.05), which proves that the drugs in each group had no effect on food intake.

[0213] Table 8 Feed intake statistics

[0214]

[0215] (3) Stool status

[0216] The feces statistics are shown in Table 9. Before medication (Day 1), the overall feces of the farm were poorly formed, with a ratio of 35% to 45% of loose feces, and obvious feed feces (accounting for 15% to 20%). On the third day of medication (Day 4), the experimental groups showed significant improvement compared with the blank group, with loose feces and feed feces decreasing by nearly 10%, especially in Experimental Groups 1 and 2, where loose feces were controlled at 20%, which was better than other experimental groups. On the fifth day of medication (Day 6), the proportion of abnormal feces in Experimental Groups 1 and 2 was controlled at about 15%, which was better than other experimental groups, and the feces were significantly improved. Continued follow-up to the second day after medication (Day 8), Experimental Groups 3 to 5 all showed rebound phenomenon, and the ratio of loose feces and feed feces increased, indicating that the control effect after stopping the medication was not ideal. In contrast, in Experimental Groups 1 and 2, loose feces were maintained at 10%, and feed feces were controlled at 5%, indicating that these two treatment plans ensured effective regulation of the intestinal tract of the chickens, and there was no recurrence, with good prevention and control effects.

[0217] Table 9 Statistical results of abnormal feces ratio

[0218]

[0219]

[0220] (4) Statistics of dead Taobao

[0221] During the experiment, the number of deaths in each group was counted every day. The results are shown in Table 10. Each group had different degrees of deaths every day. The weekly death statistics are shown in Table 10. Fig.10 The results showed that the number of deaths per week in the experimental groups was lower than that in the blank groups, especially in Experimental Group 1 and Experimental Group 2. The number of deaths during the medication period was 7 and 9 less respectively than that in the blank groups, and the mortality rate was reduced by 36.84% and 52.63%, which was a very significant decrease (P < 0.01), and was better than other experimental groups.

[0222] Table 10 Statistical results of dead Tao

[0223] Group Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Day 8 Blank Group 2 2 1 3 5 2 1 3 Test Group 1 2 1 1 0 2 3 1 2 Test Group 2 3 2 1 0 2 1 1 0 Experiment 3 groups 3 2 1 2 1 2 2 2 Test 4 groups 2 2 1 1 3 4 3 2 Test 5 groups 2 2 2 3 4 1 0 2

[0224] (5) Egg laying performance statistics

[0225] Egg laying / egg laying rate statistics: The egg laying data statistics during the trial are shown in Table 11. During the medication period, the egg laying performance of the 400-day-old sub-healthy laying hens can be improved. From the egg laying rate of about 81% before medication, the egg laying rate of the trial groups 1 and 2 increased to about 83% through intestinal regulation during the medication process, which is better than other trial groups. The summary data during the medication period are as follows Fig.11 As shown, the weekly egg production rates of Experimental Group 1 and Experimental Group 2 reached 83.01% and 83.27% respectively, which were 480 and 527 more eggs than the blank group. The egg production performance of Experimental Groups 3 to 5 also improved to a certain extent, but was lower than that of Experimental Groups 1 and 2 as a whole.

[0226] Table 11 Daily egg production / egg production rate monitoring results

[0227]

[0228] Statistics of genuine eggs: During the trial period, the number of genuine eggs, the number of defective eggs, and the rate of genuine eggs were counted daily. The results are shown in Table 12. The number of defective eggs in the blank group during the monitoring period was maintained at 220-240 eggs per day, while the number of defective eggs in the test groups 1 and 2 was maintained at 200-220 eggs per day after 3 days of medication, showing a certain decrease. In comparison, no significant improvement was found in the other test groups. The statistical results of the trial week are shown in Table 12. Fig.12 As shown, the total number of defective eggs in Experimental Group 1 and Experimental Group 2 decreased by 165 and 143 compared with the blank group, that is, the rate of genuine eggs increased by 0.17% and 0.15%, respectively. In contrast, there was no significant improvement in Experimental Groups 3 to 5 compared with the blank group.

[0229] Table 12 Daily statistics of the number (rate) of authentic eggs

[0230]

[0231] Egg quality testing: During the test period, 20 eggs were randomly selected from the genuine eggs on the 1st, 4th, 6th and 8th days, and the egg weight, eggshell strength and eggshell thickness were monitored in turn. The measurement results are shown in Table 13 and Fig.13 : The average weekly egg weight of the experimental group 1 and the experimental group 2 increased by 0.63g / egg and 0.90g / egg, the average weekly eggshell strength increased by 6.57N / egg and 9.50N / egg, and the average weekly eggshell thickness increased by 2.23mm and 0.79mm. In contrast, there was no significant improvement in the experimental groups 3 to 5, and there was no significant difference with the blank group. The improvement of egg quality is of great significance to reduce breakage in production and increase the rate of genuine eggs.

[0232] Table 13 Daily statistics of the number (rate) of authentic eggs

[0233]

[0234]

[0235] The embodiments of the present invention are described in detail above in conjunction with the accompanying drawings, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. A water-soluble cinnamon twig volatile oil solution, comprising the following components: cinnamon twig volatile oil, preservative, emulsifier, co-emulsifier and antioxidant.

2. The water-soluble cinnamon twig volatile oil solution according to claim 1, characterized in that Calculated by weight, the water-soluble cinnamon twig volatile oil solution includes 8 to 20 parts of cinnamon twig volatile oil, 0.01 to 0.5 parts of preservatives, 20 to 37 parts of emulsifiers, 20 to 45 parts of co-emulsifiers and 0.0001 to 0.7 parts of antioxidants.

3. The water-soluble cinnamon twig volatile oil solution according to claim 1, characterized in that The emulsifier includes at least one of polyoxyethylene hydrogenated castor oil and polyoxyethylene stearate; and / or, the co-emulsifier includes at least one of polyglycerol ester, glycerol, ethylene glycol, and diethylene glycol monoethyl ether; and / or, the antioxidant includes at least one of dibutyl hydroxyl radical, sodium bisulfite, and tert-butylhydroquinone; and / or, the preservative includes at least one of citric acid, sodium benzoate, and potassium sorbate.

4. The water-soluble cinnamon twig volatile oil solution according to any one of claims 1 to 3, characterized in that: The cassia twig volatile oil is prepared by the following method: Performing compound enzyme enzymolysis on the cinnamon twig aqueous solution, and collecting the enzymolysis solution; The enzymatic hydrolysate is extracted and concentrated by supercritical CO2 extraction and solvent method or distillation method to obtain cinnamon twig volatile oil; preferably, the addition amount of the complex enzyme is 2% to 4% of the mass of cinnamon twig; Preferably, the complex enzyme comprises cellulase, hemicellulase, pectinase and xylanase; Preferably, the enzyme activities of the cellulase, hemicellulase, pectinase and xylanase are 80,000 to 150,000 U / g; Preferably, the mass ratio of the cellulase, hemicellulase, pectinase and xylanase is 0.5-2:1:0.5-3:1-2; Preferably, the conditions for the complex enzyme hydrolysis treatment are pH 4.5-5.5, moisture 30%-40%, temperature 45° C.-50° C., and time 0.5 h-2 h.

5. The water-soluble cinnamon twig volatile oil solution according to claim 4, characterized in that: The conditions of the supercritical CO2 extraction are extraction pressure of 25-35 MPa, extraction temperature of 40-45°C, extraction time of 0.5-3h, and CO2 flow rate of 20-25 L / h; Preferably, the solvent in the solvent method is ethanol; Preferably, the conditions of the solvent method are that the ethanol concentration is 50% to 70%, the amount of ethanol is 10 to 20 times the weight of the medicinal material, and the extraction time is 1 to 3 hours; Preferably, the distillation method is carried out under the conditions of a distillation temperature of 88 to 100° C. and a distillation time of 3 to 5 hours.

6. The method for preparing the water-soluble cinnamon twig volatile oil solution according to any one of claims 1 to 5, comprising the following steps: Mixing an antioxidant, a preservative and an emulsifier to obtain a mixed solution A; Mixing the mixed solution A with the emulsifier to obtain a mixed solution B; The mixed solution B is mixed with cassia twig volatile oil and water, and then emulsified to obtain a water-soluble cassia twig volatile oil solution.

7. The preparation method according to claim 6, characterized in that: The emulsification conditions are as follows: pressure of 15000psi to 80000psi, temperature of 35°C to 45°C, and emulsification time of 10 to 70 minutes.

8. A medicine comprising the water-soluble cinnamon twig volatile oil solution according to any one of claims 1 to 5.

9. Use of the water-soluble cinnamon twig volatile oil solution according to any one of claims 1 to 5 and / or the drug according to claim 8 in at least one of (1) to (9): (1) Preparing products for preventing and / or treating chicken enteritis; (2) preparing products for preventing and / or treating chicken salpingitis; (3) Preparation of products for preventing and / or treating diseases caused by E. coli infection; (4) Preparation of products for preventing and / or treating diseases caused by Salmonella infection; (5) Preparation of products for preventing and / or treating diseases caused by Clostridial infections; (6) Preparing products for preventing and / or treating laying hens with clinical symptoms of sandy-shelled eggs, thin-shelled eggs, and dirty eggs; (7) Prepare products that improve egg production rate, egg quality and egg quality of laying hens; (8) Antibacterial; (9) preparing antibacterial products; Preferably, the bacteria in (8) and (9) include at least one of Staphylococcus aureus, Salmonella, Escherichia coli, and Clostridium.

10. A product comprising the water-soluble cinnamon twig volatile oil solution according to any one of claims 1 to 5 or the medicine according to claim 8.

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