Experimental dog osteoporosis model feed as well as preparation method and application thereof

Abstract

The invention relates to experimental dog osteoporosis model feed. The feed comprises 10-30 parts of puffed corn, 5-15 parts of puffed rice, 5-10 parts of puffed wheat middling, 10-15 parts of puffed soybean meal, 3-8 parts of chicken powder, 1-5 parts of beer yeast powder, 2-5 parts of corn protein powder, 1-5 parts of nata de coco, 1-3 parts of plasma protein powder, 0.04-0.08 part of choline chloride, 0.1-0.3 part of sodium chloride, 0.02-0.04 part of Dandi acid, 3-10 parts of soybean oil, 3-10 parts of duck oil, 0.5-1 part of stone powder, 1-4 parts of chicken liver powder and 4-8 parts of a beef flavor agent. The multi-vitamin multi-mineral premix can also comprise 2-4 parts of bismuth potassium citrate, so that the problem of stomach discomfort is solved. The feed prepared by the optimized preparation method of the feed is used for feeding experimental dogs, and the dog osteoporosis animal model is successfully established in a short time by a method of reducing the content of calcium in the dog feed and combining glucocorticoids.

Description

Technical Field

[0001] The invention relates to the technical field of animal model feed, and in particular to an experimental dog osteoporosis model feed and a preparation method and application thereof. Background Art

[0002] Osteoporosis (hereinafter also referred to as "OP") is a systemic skeletal disease characterized by decreased bone mass, destruction of bone microstructure, and decreased bone strength, leading to increased bone fragility and susceptibility to fractures.

[0003] Modern medicine divides osteoporosis into three categories: primary, secondary, and idiopathic osteoporosis. Primary osteoporosis is caused by a sudden decrease in sex hormones in the body due to age and physiological degenerative changes. It is divided into type I postmenopausal osteoporosis and type II senile osteoporosis. Secondary osteoporosis is induced by diseases or drug factors, such as endocrine metabolic diseases (diabetes, hyperthyroidism), kidney disease, liver disease, etc., and drug-induced osteoporosis is caused by long-term high-dose heparin, immunosuppressants, anti-epileptic drugs, and glucocorticoids. Idiopathic osteoporosis is generally accompanied by a history of genetic diseases and is more common in women. Osteoporosis in women during lactation and pregnancy is often also classified as this category.

[0004] At present, there are many methods to induce osteoporosis models, which can be divided into surgical induction (ovariectomy, parathyroidectomy, pinealectomy, etc.), drug induction (such as retinoic acid, glucocorticoids, etc.), dietary induction (such as low calcium, phosphorus, vitamin D diet, etc.), disuse induction (such as tail suspension, fixation, etc.), combined modeling (such as surgery combined with low calcium diet, low calcium diet combined with glucocorticoids, etc.). Nutrition is one of the many factors that affect bone density. By restricting the intake of calcium, vitamin D, protein, etc. in the diet, a nutrient deficiency osteoporosis model can be successfully established. This model is of great significance for studying osteoporosis caused by nutritional deficiencies. However, due to the complex feed formula and many influencing factors, it is difficult to popularize and promote. At the same time, because the single application of nutrition method modeling takes a long time and has a low success rate, it is often used as an auxiliary method. The use of low calcium feed and hormones (by intramuscular injection) to jointly construct an osteoporosis animal model is a commonly used method at present. It has the advantages of simple operation and good repeatability, but it also has the disadvantages of long modeling time, many animal side effects, and high experimental costs. Animal models are needed to study osteoporosis. Animal models often use rats, mice, rabbits, pigs, dogs, sheep and primates as research subjects. Rats and mice are widely studied as osteoporosis animal models, but they are not suitable for surgical experiments due to their small size. Among large animals, the sheep osteoporosis model is more thoroughly studied, while there are few studies on the canine osteoporosis animal model at home and abroad. Summary of the invention

[0005] In order to overcome at least one problem existing in the prior art, the present invention provides an experimental canine osteoporosis model feed, which is applied to constructing a canine osteoporosis model by optimizing its raw material composition and various preparation steps, so as to solve the problems existing in the related art such as complex modeling method, difficult operation, and long modeling time.

[0006] To achieve the above object, the present invention adopts the following technical solution:

[0007] The first aspect of the present invention is to provide an experimental canine osteoporosis model feed, which is an experimental low-calcium dog food, and dexamethasone is sprayed on the outside of the experimental low-calcium dog food.

[0008] Furthermore, the feed comprises, by weight: 10-30 parts of puffed corn, 5-15 parts of puffed rice, 5-10 parts of puffed second flour, 10-15 parts of puffed soybean meal, 3-8 parts of chicken meal, 1-5 parts of brewer's yeast powder, 2-5 parts of corn protein powder, 1-5 parts of coconut fiber energy, 1-3 parts of plasma protein powder, 0.04-0.08 parts of choline chloride, 0.1-0.3 parts of sodium chloride, 0.02-0.04 parts of datian acid, 3-10 parts of soybean oil, 3-10 parts of duck oil, 0.5-1 parts of stone powder, 1-4 parts of chicken liver powder, 4-8 parts of beef flavoring agent, and 2-4 parts of multivitamin and mineral premix; wherein, the feed also comprises glucocorticoid, and 80-120 mg of dexamethasone is added to each kg of feed.

[0009] The dosage of dexamethasone is determined by the following criteria: Injection of 2.5 mg / kg / day of dexamethasone into mice can induce osteoporosis model. Normal adult dogs weigh about 10 kg. Based on the daily food intake of 200-300 g per dog, the amount of dexamethasone added to dog food is 80-120 mg / kg.

[0010] Furthermore, the feed comprises, by weight: 20-30 parts of puffed corn, 8-12 parts of puffed rice, 8-10 parts of puffed flour, 12-15 parts of puffed soybean meal, 5-8 parts of chicken meal, 1-3 parts of brewer's yeast powder, 2-4 parts of corn protein powder, 1-3 parts of coconut fiber energy, 2-3 parts of plasma protein powder, 0.05-0.07 parts of choline chloride, 0.2-0.3 parts of sodium chloride, 0.03-0.04 parts of datian acid, 5-10 parts of soybean oil, 5-10 parts of duck oil, 0.7-1 parts of stone powder, 1-3 parts of chicken liver powder, 4-6 parts of beef flavoring agent, and 3-4 parts of multivitamin and multi-mineral premix; wherein, the feed also comprises glucocorticoid, and 90-110 mg of dexamethasone is added to each kg of feed.

[0011] Furthermore, the feed includes, by weight: 25 parts of puffed corn, 10 parts of puffed rice, 9 parts of puffed flour, 14 parts of puffed soybean meal, 7 parts of chicken meal, 2 parts of brewer's yeast powder, 3 parts of corn protein powder, 1 part of coconut fiber energy, 2.8 parts of plasma protein powder, 0.06 parts of choline chloride, 0.3 parts of sodium chloride, 0.04 parts of datian acid, 7 parts of soybean oil, 7 parts of duck oil, 0.8 parts of stone powder, 2 parts of chicken liver powder, 5 parts of beef flavoring agent, and 4 parts of multivitamin mineral premix; wherein, the feed also includes glucocorticoids, and 100 mg of dexamethasone is added to each kg of feed.

[0012] Furthermore, the feed is a low-calcium dog food with an unbalanced calcium-phosphorus ratio, and its calcium content is low, while the phosphorus content meets the requirements. In order to ensure that the calcium content in the dog food is low and other nutritional indicators are within the national standard range, the chicken meal content is reduced and the plasma protein powder is increased, and the calcium hydrogen phosphate in the feed is replaced with sodium dihydrogen phosphate in equal proportions to ensure that the phosphorus content in the feed meets the national standard requirements while the calcium content is low.

[0013] Furthermore, the feed also includes: 2-4 parts of potassium bismuth citrate; preferably, 3 parts of potassium bismuth citrate. Potassium bismuth citrate is first sprayed on the outside of the low-calcium dog food, and then dexamethasone is sprayed. Long-term use of dexamethasone may cause excessive secretion of pepsin and gastric acid, leading to symptoms of stomach discomfort such as acid reflux, nausea, vomiting, and even gastric ulcers. Potassium bismuth citrate tablets, as a gastric mucosal protective agent, can produce precipitation under gastric acid conditions, effectively preventing gastric acid and the like from invading the ulcer surface; at the same time, it can also reduce the activity of pepsin and protect the gastric mucosa, thereby improving the external absorption environment of dexamethasone.

[0014] Furthermore, the multivitamin and multi-mineral premix is ​​composed of 1% complex vitamin premix, 1.5% complex trace element premix, 1% lysine, 2% taurine, 5% methionine and carrier powder, and mainly contains VE 1600IU / kg, VB1380mg / kg, VB2 230mg / kg, niacin 1200mg / kg, biotin 4mg / kg, folic acid 50mg / kg, choline 500mg / kg, VB6200mg / kg, VB 12 595mg / kg, VA 300000IU / kg, VD72000 IU / kg, iron 2.3g / kg, copper 35mg / kg, manganese 655mg / kg, zinc 1000mg / kg, selenium 1.4mg / kg.

[0015] The second aspect of the present invention is to provide a method for preparing an experimental canine osteoporosis model feed as described in any one of the first aspects, comprising the following steps:

[0016] S1. Put the puffed corn, puffed flour, puffed soybean meal and puffed rice in predetermined weight portions from the feeding port in sequence and put them into the silos of different raw materials for standby use; grind the raw materials and mix the grinded raw materials evenly to obtain a first mixed material;

[0017] S2, mixing the first mixture with chicken meal, coconut fiber, brewer's yeast powder, corn protein powder, plasma protein powder, stone powder, sodium chloride, Dadi acid, choline chloride, and multivitamin premix in predetermined weight portions to obtain a second mixture;

[0018] S3, subjecting the second mixed material to tempering treatment, and adding a predetermined weight portion of duck fat and soybean oil during the tempering process; and subjecting the tempered raw material to puffing and granulation to obtain a third mixed material;

[0019] S4. Vacuum spraying the third mixed material with predetermined weight portions of beef flavor, chicken liver powder, and dexamethasone in sequence to obtain the feed; wherein, when bismuth potassium citrate is contained, vacuum spraying the third mixed material with predetermined weight portions of beef flavor, chicken liver powder, bismuth potassium citrate, and dexamethasone in sequence to obtain the feed.

[0020] Furthermore, in step S1, each raw material is crushed to a fineness of 0.25 mm for mixing and granulation.

[0021] Furthermore, in step S2, the secondary mixing needs to ensure that the nutrient content in the feed is consistent.

[0022] Furthermore, in step S3, the conditioning treatment is: conditioning at 125° C. and 0.5 MPa steam to pre-gelatinize starch, denature protein, and soften raw materials, so as to improve the puffing granulation effect and quality.

[0023] Furthermore, in step S3, the puffing granulation is: extruding into triangular granules with a 7 mm die, drying at 110° C. for 15 min and drying at 85° C. for 10 min after molding.

[0024] Furthermore, the dog food after the spraying step is cooled and then packed with nitrogen.

[0025] The third aspect of the present invention is to provide an application of the experimental canine osteoporosis model feed described in any one of the first aspects, or the experimental canine osteoporosis model feed prepared by any one of the preparation methods described in the second aspect; wherein the feed is used to construct an experimental canine osteoporosis model.

[0026] The fourth aspect of the present invention is to provide a method for constructing an experimental canine osteoporosis model, which comprises the steps of: using any experimental canine osteoporosis model feed described in the first aspect or an experimental canine osteoporosis model feed prepared by any preparation method described in the second aspect, feeding the experimental dog twice a day (once in the morning and once in the afternoon), the total feeding amount is 2.5% of the experimental dog's body weight, and the modeling period is 8 to 12 weeks.

[0027] Furthermore, in the above construction method, the rats are fed with feed sprayed with dexamethasone or feed sprayed with potassium bismuth citrate + dexamethasone during the modeling period.

[0028] Furthermore, in the above construction method, the feed sprayed with dexamethasone is fed in the first half of the modeling cycle, and the feed sprayed with potassium bismuth citrate + dexamethasone is fed in the second half of the modeling cycle.

[0029] Further, the experimental dogs are healthy female beagles aged 5-7 years. From the perspective of physiological characteristics, female animals may be more prone to osteoporosis after ovariectomy, and thus are more suitable for constructing an osteoporosis model. The ratio of cortical bone to cancellous bone, the Haversian system, and the morphology of cancellous bone reconstruction in beagles are similar to those in humans, which makes them an ideal model for studying osteoporosis. The selection of healthy dogs that meet the experimental requirements is to ensure that they have no symptoms of osteoporosis before the experiment, so the present invention uses healthy female beagles as experimental animals for establishing an osteoporosis model.

[0030] Compared with the prior art, the present invention adopts the above technical solution to have the following beneficial effects:

[0031] The low calcium dog food prepared by the present invention meets GB / T 14924.1 and GB / T 14924.2, and except that the calcium content is lower than the national standard requirement, other nutritional indicators meet GB14924.3. By changing the nutritional composition of the feed and optimizing the preparation process, an osteoporosis model caused by the combination of nutritional deficiency and glucocorticoid dexamethasone is achieved.

[0032] The present invention uses puffing raw materials to pre-ripen cereal raw materials such as corn, second-grade flour, soybean meal and rice. Through pretreatment, the molecular structure of the raw materials will change, making it easier to form a porous and crispy structure during the puffing process. The puppy food of the present invention is packaged in nitrogen, which can effectively remove oxygen in the package and slow down the oxidation process of the dog food, thereby extending the shelf life of the dog food. Nitrogen packaging can also reduce and delay the loss of flavor and nutrients, and maintain the freshness and nutritional value of the dog food. In addition, nitrogen packaging can reduce physical damage to the dog food during the packaging process, such as extrusion, friction, etc., thereby maintaining the integrity and shape of the dog food.

[0033] The present invention sprays dexamethasone or potassium bismuth citrate + dexamethasone externally after the material is dried, and the modeling is combined by feeding low-calcium dog food plus dexamethasone, which is simple and convenient; the oral dexamethasone method is adopted, which avoids the side effects such as wound infection and skin changes caused by intramuscular injection of dexamethasone, and its safety is relatively better. At the same time, the external spraying of dexamethasone can also avoid the influence of drug failure or adverse consequences caused by high temperature and high pressure. In addition, in order to avoid the gastrointestinal mucosal barrier function that may be caused by long-term use of dexamethasone, the present invention also adds potassium bismuth citrate in the feed, which can help repair the gastric mucosa as a gastric mucosal protective agent, improve the absorption environment, thereby indirectly promoting the absorption of dexamethasone, so as to build a dog osteoporosis model more safely and quickly. DETAILED DESCRIPTION

[0034] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without creative work belong to the scope of protection of the present invention. The experimental methods without specific conditions in the following embodiments are usually measured according to national standards. The experimental materials without the source in the following embodiments are all commercially available raw materials. The equipment used in each step in the following embodiments is conventional equipment. If there is no corresponding national standard, it is carried out according to the general international standards, conventional conditions, or according to the conditions recommended by the manufacturer. Unless otherwise specified, all parts are weight parts and all percentages are mass percentages. Unless otherwise defined or specified, all professional and scientific terms used in the present invention have the same meaning as those familiar to those skilled in the art. In addition, any method and material similar or equal to the recorded content can be applied to the method of the present invention.

[0035] In certain embodiments of the present invention, the low-calcium dog food of the present invention comprises the following raw materials in parts by weight: 10-30 parts of puffed corn, 5-15 parts of puffed rice, 5-10 parts of puffed second flour, 10-15 parts of puffed soybean meal, 3-8 parts of chicken meal, 1-5 parts of brewer's yeast powder, 2-5 parts of corn protein powder, 1-5 parts of coconut fiber energy, 1-3 parts of plasma protein powder, 0.04-0.08 parts of choline chloride, 0.1-0.3 parts of sodium chloride, 0.02-0.04 parts of great acid, 3-10 parts of soybean oil, 3-10 parts of duck oil, 0.5-1 parts of stone powder, 1-4 parts of chicken liver powder, 4-8 parts of beef flavor, and 2-4 parts of multivitamin and multi-mineral premix; wherein the feed also includes glucocorticoids, and 80-120 mg of dexamethasone is added to each kg of feed; the above-mentioned low-calcium dog food may also include: 2-4 parts of potassium bismuth citrate. The low-calcium dog diets prepared within the above-mentioned component ratio range can successfully establish a canine osteoporosis model.

[0036] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments can be combined with each other. The present invention is further described below in conjunction with specific embodiments, but is not intended to be a limitation of the present invention.

[0037] Example 1 - Preparation of low calcium dog food

[0038] This embodiment is a composition and preparation of some preferred low-calcium dog food, and the feed specifically includes:

[0039] (1) Low calcium dog food 1: 25 parts of puffed corn, 10 parts of puffed rice, 9 parts of puffed flour, 14 parts of puffed soybean meal, 7 parts of chicken meal, 2 parts of brewer's yeast powder, 3 parts of corn gluten powder, 1 part of coconut fiber, 2.8 parts of plasma protein powder, 0.06 parts of choline chloride, 0.3 parts of sodium chloride, 0.04 parts of dacin, 7 parts of soybean oil, 7 parts of duck fat, 0.8 parts of stone powder, 2 parts of chicken liver powder, 5 parts of beef flavor, 4 parts of multivitamin and multimineral premix.

[0040] (2) Low calcium dog food 2: 25 parts of puffed corn, 10 parts of puffed rice, 9 parts of puffed flour, 14 parts of puffed soybean meal, 7 parts of chicken meal, 2 parts of brewer's yeast powder, 3 parts of corn protein powder, 1 part of coconut fiber, 2.8 parts of plasma protein powder, 0.06 parts of choline chloride, 0.3 parts of sodium chloride, 0.04 parts of dachinic acid, 7 parts of soybean oil, 7 parts of duck fat, 0.8 parts of stone powder, 2 parts of chicken liver powder, 5 parts of beef flavoring agent, and 4 parts of multivitamin and mineral premix; wherein the low calcium dog food also includes glucocorticoids, and 100 mg of dexamethasone is added to each kg of low calcium dog food.

[0041] (3) Low calcium dog food 3: 25 parts of puffed corn, 10 parts of puffed rice, 9 parts of puffed flour, 14 parts of puffed soybean meal, 7 parts of chicken meal, 2 parts of brewer's yeast powder, 3 parts of corn protein powder, 1 part of coconut fiber, 2.8 parts of plasma protein powder, 0.06 parts of choline chloride, 0.3 parts of sodium chloride, 0.04 parts of datian acid, 7 parts of soybean oil, 7 parts of duck fat, 0.8 parts of stone powder, 2 parts of chicken liver powder, 5 parts of beef flavor, 4 parts of multivitamin mineral premix, and 3 parts of potassium bismuth citrate; wherein the low calcium dog food also includes glucocorticoids, and 100 mg of dexamethasone is added to each kg of low calcium dog food.

[0042] In the above low calcium dog food, the multivitamin multi-mineral premix is ​​composed of 1% complex vitamin premix, 1.5% complex trace element premix, 1% lysine, 2% taurine, 5% methionine and carrier powder, mainly containing VE 1600IU / kg, VB1380mg / kg, VB2 230mg / kg, niacin 1200mg / kg, biotin 4mg / kg, folic acid 50mg / kg, choline 500mg / kg, VB6200mg / kg, VB 12595mg / kg, VA 300000IU / kg, VD72000 IU / kg, iron 2.3g / kg, copper 35mg / kg, manganese 655mg / kg, zinc 1000mg / kg, selenium 1.4mg / kg.

[0043] A preferred preparation process of the above experimental dog food includes the following steps:

[0044] Preparation of feed: The production process of experimental dog food mainly consists of raw material receiving and cleaning, coarse crushing, batching, mixing, expansion, secondary crushing, secondary mixing, conditioning, puffing, drying, spraying, cooling, transportation, grading, cleaning, finished product packaging and other processes, adopting the process of crushing first and then mixing.

[0045] Before receiving the feed raw materials, they are first sampled and inspected to ensure that their nutritional content, color, and taste meet the acceptance standards and are free of mold, etc. They are lifted to the raw material bin by a bucket elevator through the feeding port and pass through a primary cleaning screen and a permanent magnetic cylinder before entering the raw material bin to remove lint, dust, gravel, iron filings and other debris. It is understandable that the total amount of feed prepared can be adjusted according to the actual production capacity of the equipment, but the weight fraction of each component remains unchanged.

[0046] The automatically weighed corn, second-grade flour, soybean meal and rice are crushed separately. The particle size of the crushed material is 0.25mm. Each crushed raw material is sent to the extruder for pre-ripening. The extruder has a pair of screws and screw sleeves. After the mixed material enters the extrusion chamber, it is squeezed, rubbed, sheared, etc. between the screw sleeves, and its internal pressure continues to rise, up to 4MPA, and the temperature continues to rise, up to 130℃. Within 3-7 minutes, the temperature and pressure rise sharply, the organizational structure of the raw material changes, the crude fiber is destroyed, and harmful bacteria such as Salmonella are killed. The high-temperature and high-pressure raw materials come out of the discharge port, and their pressure is suddenly released, and the water release part flashes. After cooling, the material has a loose and porous structure. The whole pre-ripening process takes 10 minutes. The ripened raw materials are puffed corn, puffed second-grade flour, puffed soybean meal, and puffed rice.

[0047] The puffed corn, puffed second-grade flour, puffed soybean meal, and puffed rice are crushed again: they are put into the feeding port in sequence and enter the silos of different raw materials for standby; each raw material is crushed to a fineness of 0.25mm for mixing and granulation; the crushed raw materials are mixed evenly to obtain the first mixture. The remaining raw material components, chicken powder, coconut fiber energy, brewer's yeast powder, corn protein powder, plasma protein powder, stone powder, sodium chloride, Dadi acid, choline chloride, and multivitamin premix are added for secondary mixing to ensure that the nutrient content of each grain of dog food is consistent, and the second mixture is obtained by mixing for 2.5 minutes. The second mixture is tempered at 125℃ and 0.5Mpa steam to pre-gelatinize starch, denature protein, and soften the raw materials, which is conducive to improving the puffing granulation effect and quality; duck oil and soybean oil are added during the tempering process; the tempered raw materials are puffed and granulated, extruded into triangular granules with a 7mm mold, and dried at 110℃ for 15min and 85℃ for 10min after molding to obtain the third mixture. The third mixed material is vacuum sprayed with beef flavor, chicken liver powder, bismuth potassium citrate and dexamethasone in sequence to obtain low calcium dog food, which is then packaged with nitrogen after cooling.

[0048] Example 2- Construction of osteoporosis model

[0049] This example is a construction of some preferred experimental canine osteoporosis models, which uses the feed prepared in Example 1 and Example 2, and specifically includes the following steps:

[0050] (1) Experimental materials and main instruments: Dexamethasone was produced by Hubei Tianyao Pharmaceutical Co., Ltd., ketamine mixture was produced by Hengrui Medicine, dual-energy X-ray absorptiometry was produced by a French company, Hitachi 7060 fully automatic biochemical analyzer, biomechanical testing machine, and ultra-low temperature refrigerator (-80°C).

[0051] (2) Experimental animals: 30 healthy female beagle dogs aged 5-7 years weighing about 10 kg were purchased and selected. After purchase, they were adaptively fed for 1 week and randomly divided into 6 groups (5 dogs in each group). The cycle was 12 weeks, and the dogs were fed once in the morning and afternoon, and the total amount of feeding was 2.5% of the dog's body weight.

[0052] (3) Experimental group settings:

[0053] Control group 1: fed with ordinary dog ​​maintenance feed (Jiangsu Collaborative Bioengineering (Yangzhou) Co., Ltd., catalog number: XTC03WC-001);

[0054] Control group 2: fed with ordinary dog ​​maintenance feed (XTC03WC-001) sprayed with dexamethasone;

[0055] Experimental group 1: fed with low-calcium feed 1 in Example 1 (low-calcium feed without dexamethasone);

[0056] Experimental group 2: fed with low-calcium feed 2 in Example 1 (low-calcium feed sprayed with dexamethasone);

[0057] Experimental group 3: fed with low-calcium feed 3 in Example 1 (low-calcium feed sprayed with bismuth potassium citrate and dexamethasone);

[0058] Experimental Group 4: The mice were fed with low-calcium feed 2 in Example 1 (low-calcium feed sprayed with dexamethasone) for the first 6 weeks and low-calcium feed 3 in Example 1 (low-calcium feed sprayed with bismuth potassium citrate and dexamethasone) for the next 6 weeks.

[0059] Starting from the 4th week, the total number of vomiting times of the dogs was counted every 2 weeks; bone density measurement and biochemical examination were performed before the experiment, 4th week, and 8th week of the experiment, and bone density measurement, biochemical examination, bone biomechanics and bone tissue morphology examination were performed at 12th week of the experiment. Specific test data are shown in Table 1-5.

[0060] Table 1 - Statistics of total vomiting times in dogs

[0061] Group Experimental 4 weeks Experimental 6 weeks Experimental 8 weeks Experimental 10 weeks Experimental 12 weeks Control group 1 0 0 0 0 0 Control group 2 0 1 2 5 10 Experimental Group 1 0 0 0 0 0 Experimental Group 2 0 1 3 6 12 Experimental Group 3 0 0 1 2 4 Experimental Group 4 0 1 2 5 8

[0062] From the total number of vomiting times of dogs counted in Table 1, it can be seen that the consumption of dexamethasone can cause different degrees of stomach discomfort, thereby inducing vomiting, but no dog death was caused in the above experiments, which shows that the feed prepared by the present invention has a certain degree of safety; from the results of experimental groups 3 and 4, it can be seen that after adding potassium bismuth citrate, the vomiting phenomenon can be effectively improved, which is conducive to the absorption of dog food and dexamethasone.

[0063] Table 2 - Comparison of BMD in dogs (g / cm 2 )

[0064]

[0065]

[0066] From the BMD statistics of dogs in Table 2, it can be seen that there was no statistical difference in bone density among the six groups before the experiment; bone density began to decrease after 8 weeks of adding dexamethasone alone or using low-calcium dog food alone, and the bone density decreased significantly after 12 weeks; however, the bone density of experimental groups 2 to 4 using low-calcium dog food combined with dexamethasone began to decrease from 4 weeks, and the bone density decreased significantly after 8 and 12 weeks. Among them, the bone density of experimental groups 3 and 4 added with potassium bismuth citrate at 8 weeks was equivalent to or slightly higher than that of other groups at 12 weeks, and their modeling effect was better.

[0067] Table 3 - Comparison of blood calcium, blood phosphorus and alkaline phosphatase (AKP) in dogs

[0068]

[0069] From the statistical data of dog blood calcium, blood phosphorus and alkaline phosphatase in Table 3, it can be seen that biochemical examination found that there was no significant difference in blood calcium among the six groups before and after the experiment; the blood phosphorus of the control group 2 with dexamethasone alone, the experimental group 2 with low calcium dog food combined with dexamethasone, and the experimental group 3 and experimental group 4 with low calcium dog food combined with dexamethasone added with potassium citrate bismuth increased from 4 weeks, and further increased at 8 weeks and 12 weeks, and there was no significant change in experimental group 1 and control group 1; compared with control group 1, the alkaline phosphatase of the experimental groups increased significantly at 4, 8 and 12 weeks of the experiment when dexamethasone was added alone or low calcium dog food was used alone, but the increase was more significant in the combination of low calcium dog food combined with dexamethasone (with or without potassium citrate bismuth), among which the effect of feed containing potassium citrate bismuth was more significant.

[0070] Table 4 – Comparison of canine biomechanics

[0071]

[0072] From the canine biomechanics statistics in Table 4, it can be seen that the results of bone biomechanics examination at the 12th week of the experiment showed that compared with the control group 1, the lumbar vertebrae compression limit load, ultimate strength and compression modulus of the dogs that added dexamethasone alone or used low-calcium dog food alone were significantly lower than those of the control group 1; compared with the control group 1, the femoral three-point bending limit load of the dogs that added dexamethasone alone or used low-calcium dog food alone was significantly lower than that of the control group 1; the experimental group was lower than the control group 1, but the combination of low-calcium dog food and dexamethasone (with or without bismuth potassium citrate) had a more significant reduction, among which the effect of the feed containing bismuth potassium citrate was more significant.

[0073] Table 5 – Comparison of trabecular bone structure in dogs

[0074] Group Tb.Th(mm) Tb.N(1 / mm) Tb.Sp(mm) Tb..Ar(%) Control group 1 1.035±0.038 3.13±0.18 2.57±0.19 41.08±3.21 Control group 2 0.918±0.023 2.93±0.12 2.63±0.27 32.31±7.23 Experimental Group 1 0.976±0.081 2.99±0.17 2.74±0.23 34...89±4.21 Experimental Group 2 0.829±0.042 2.62±0.16 2.98±0.22 28.92±4.93 Experimental Group 3 0.798±0.035 2.48±0.15 3.19±0.23 26.49±4.58 Experimental Group 4 0.806±0.039 2.53±0.14 3.07±0.25 27.05±4.75

[0075] From the statistics of canine trabecular structure in Table 4, it can be seen that at week 12, compared with control group 1, the addition of dexamethasone alone or the use of low-calcium dog food alone did not significantly change the trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular separation (Tb.Sp), and the trabecular area percentage (Tb.Ar) was significantly reduced. The trabecular area percentage (Tb.Ar), trabecular thickness (Tb.Th), and trabecular number (Tb.N) of experimental groups 2 to 4 of the low-calcium dog food combined with dexamethasone combination (with or without potassium bismuth citrate) were significantly reduced compared with the control group, and the trabecular separation (Tb.Sp) was significantly increased compared with the control group, and the effect of the feed containing potassium bismuth citrate was more significant.

[0076] The above examples confirm from multiple aspects such as bone density, mechanical experiments and histological examinations that the application of a low-calcium diet combined with dexamethasone in experimental dogs can better construct an osteoporosis model, causing significant degeneration of bone structure, reduction of bone mass, and decrease of bone strength, resulting in osteoporosis similar to that in humans. Moreover, after adding potassium bismuth citrate, the safety is better and the modeling effect is better.

[0077] As can be seen from the above embodiments, the present invention sprays the glucocorticoid dexamethasone externally in low-calcium dog food and feeds it to experimental dogs, thereby realizing the construction of an osteoporosis model. In addition, potassium bismuth citrate is used to improve the stomach discomfort of experimental dogs, improve the absorption environment of feed and hormones, and construct an osteoporosis model more safely and quickly. The construction method involved in the present invention is also innovative, unique and simple, and provides a new and more effective experimental means for osteoporosis research. The successful establishment of the canine osteoporosis model constructed by the invention will provide a basis for surgical experimental research, which is conducive to promoting the development of new surgical methods.

[0078] The specific embodiments of the present invention are described in detail above, but they are only examples, and the present invention is not limited to the specific embodiments described above. For those skilled in the art, any equivalent modifications and substitutions made to the present invention are also within the scope of the present invention. Therefore, the equalization changes and modifications made without departing from the spirit and scope of the present invention should be included in the scope of the present invention.

Claims

1. An experimental dog osteoporosis model feed, characterized in that: The feed comprises, by weight, 10-30 parts of puffed corn, 5-15 parts of puffed rice, 5-10 parts of puffed second flour, 10-15 parts of puffed soybean meal, 3-8 parts of chicken meal, 1-5 parts of brewer's yeast powder, 2-5 parts of corn protein powder, 1-5 parts of coconut fiber energy, 1-3 parts of plasma protein powder, 0.04-0.08 parts of choline chloride, 0.1-0.3 parts of sodium chloride, 0.02-0.04 parts of datian acid, 3-10 parts of soybean oil, 3-10 parts of duck oil, 0.5-1 parts of stone powder, 1-4 parts of chicken liver powder, 4-8 parts of beef flavoring agent, and 2-4 parts of multivitamin and multi-mineral premix; wherein, the feed also comprises glucocorticoid, and 80-120 mg of dexamethasone is added to each kg of feed.

2. The experimental dog osteoporosis model feed according to claim 1, characterized in that: The feed comprises, by weight, 25 parts of puffed corn, 10 parts of puffed rice, 9 parts of puffed flour, 14 parts of puffed soybean meal, 7 parts of chicken meal, 2 parts of brewer's yeast powder, 3 parts of corn protein powder, 1 part of coconut fiber energy, 2.8 parts of plasma protein powder, 0.06 parts of choline chloride, 0.3 parts of sodium chloride, 0.04 parts of datian acid, 7 parts of soybean oil, 7 parts of duck oil, 0.8 parts of stone powder, 2 parts of chicken liver powder, 5 parts of beef flavoring agent, and 4 parts of multivitamin and mineral premix; wherein, the feed also comprises glucocorticoid, and 100 mg of dexamethasone is added to each kg of feed.

3. The experimental dog osteoporosis model feed according to claim 1, characterized in that: The feed is a low-calcium dog food with an unbalanced calcium-phosphorus ratio, with a low calcium content and a phosphorus content that meets the requirements.

4. The experimental dog osteoporosis model feed according to claim 1, characterized in that: The feed also includes: 2-4 portions of bismuth potassium citrate.

5. The experimental dog osteoporosis model feed according to claim 1, characterized in that: The multivitamin and multimineral premix is ​​composed of 1% compound vitamin premix, 1.5% compound trace element premix, 1% lysine, 2% taurine, 5% methionine and carrier powder, and mainly contains VE 1600IU / kg, VB1 380mg / kg, VB2 230mg / kg, niacin 1200mg / kg, biotin 4mg / kg, folic acid 50mg / kg, choline 500mg / kg, VB6 200mg / kg, VB 12 595mg / kg, VA 300000IU / kg, VD72000IU / kg, iron 2.3g / kg, copper 35mg / kg, manganese 655mg / kg, zinc 1000mg / kg, selenium 1.4mg / kg.

6. A method for preparing an experimental canine osteoporosis model feed as claimed in any one of claims 1 to 5, characterized in that: The following steps are involved: S1. Put the puffed corn, puffed flour, puffed soybean meal and puffed rice in predetermined weight portions from the feeding port in sequence and put them into the silos of different raw materials for standby use; grind the raw materials and mix the grinded raw materials evenly to obtain a first mixed material; S2, mixing the first mixture with chicken meal, coconut fiber, brewer's yeast powder, corn protein powder, plasma protein powder, stone powder, sodium chloride, Dadi acid, choline chloride, and multivitamin premix in predetermined weight portions to obtain a second mixture; S3, subjecting the second mixed material to tempering treatment, and adding a predetermined weight portion of duck fat and soybean oil during the tempering process; and subjecting the tempered raw material to puffing and granulation to obtain a third mixed material; S4. Vacuum spraying the third mixed material with predetermined weight portions of beef flavor, chicken liver powder, and dexamethasone in sequence to obtain the feed; wherein, when bismuth potassium citrate is contained, vacuum spraying the third mixed material with predetermined weight portions of beef flavor, chicken liver powder, bismuth potassium citrate, and dexamethasone in sequence to obtain the feed.

7. The method for preparing the experimental canine osteoporosis model feed according to claim 6, characterized in that: In step S1, each raw material is crushed to a fineness of 0.25 mm for mixing and granulation; In step S2, the secondary mixing needs to ensure that the nutrient content in the feed is consistent; In step S3, the conditioning treatment is: conditioning at 125° C. and 0.5 MPa steam to pre-gelatinize starch, denature protein, and soften raw materials, so as to improve the puffing granulation effect and quality; In the step S3, the puffing granulation is: extruding into triangular granules with a 7 mm die, drying at 110° C. for 15 min and drying at 85° C. for 10 min after forming.

8. A use of the experimental canine osteoporosis model feed according to any one of claims 1 to 5, or the experimental canine osteoporosis model feed prepared by the preparation method according to any one of claims 6 to 7, characterized in that: The feed was used to construct an experimental canine osteoporosis model.

9. A method for constructing an experimental canine osteoporosis model, characterized in that: The construction method comprises the steps of: using the experimental dog osteoporosis model feed as described in any one of claims 1 to 5 or the experimental dog osteoporosis model feed prepared by the preparation method as described in any one of claims 6 to 7, feeding the experimental dog twice a day, the total feeding amount is 2.5% of the body weight of the experimental dog, and the modeling period is 8 to 12 weeks.

10. The construction method according to claim 9, characterized in that: The experimental dogs were healthy female beagles aged 5-7 years.

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