Preparation for diarrhea molding as well as preparation method and application thereof
By enzymatically treated senna, buckthorn and Atractylodesis, the diarrhea modeling preparations were solved, and the traditional diarrhea model was cumbersome and poor stability were achieved, and efficient and stable modeling and palatability were improved.
Patent Information
- Application Number
- CN202411924587.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-25
- Publication Date
- 2025-04-25
AI Technical Summary
The construction of traditional diarrhea models is complicated, which can easily cause esophageal damage and poor stability.
After mixing senna, buckthorn and Atractylodesis, heating and sonication, multiple enzymes were added for enzymatic decomposition, followed by filtration, distillation and drying, a preparation for diarrhea modeling was obtained and fed to the animals in mixture with the basic diet.
It reduces the irritation of medicinal materials, improves the palatability of the preparation, avoids constipation problems, achieves efficient and stable modeling, reduces the risk of animal death, and simplifies the operation process.
Smart Images
Figure CN119868437A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of animal model experiments, and specifically relates to a preparation for diarrhea modeling, a preparation method and an application thereof. Background Art
[0002] Diarrhea is a common physiological or pathological reaction in livestock and poultry, with watery feces, rapid dehydration and autointoxication as the main symptoms. It is a clinical disease with great harm. As a scientific research leader, animal experiments can make up for the shortcomings of clinical research, especially in clinical research involving ethical issues and some biased clinical research. Therefore, studying animal models with good objectivity, recognition and repeatability has a positive effect on understanding the pathogenesis, pathophysiology and drug treatment.
[0003] Traditional diarrhea models are often constructed by gavage with senna leaves. Senna leaves are the dried leaflets of the small shrub plants of the Leguminosae family, Senna sennae or Senna interlobata. It is a commonly used irritant laxative with a history of more than a thousand years of use. It mainly acts on the large intestine and has the ability to produce contractions and enhance colon motility through the sennosides it contains. After reaching the intestine, sennosides are decomposed into aglycones by bacteria and enzymes, stimulating the large intestine, increasing tension and peristalsis, and reducing water absorption to cause diarrhea. The ethanol extract of senna leaves has a significant inhibitory effect on a variety of bacteria such as Escherichia coli, Staphylococcus aureus, and Salmonella typhimurium. However, senna leaves often cause a variety of adverse reactions, such as nausea, vomiting, abdominal pain, diarrhea, gastrointestinal bleeding, jaundice, and allergies. However, long-term or high-dose use of senna leaves may weaken gastrointestinal motility, leading to spleen deficiency, difficulty in defecation, and aggravated constipation. In addition, traditional senna leaf gavage is cumbersome, and manual operation can easily cause esophageal damage. Summary of the invention
[0004] The purpose of the present invention is to overcome the problems of the diarrhea model in the prior art, such as complicated operation, easy to cause esophageal damage, poor stability, etc.
[0005] To this end, the present invention provides a method for preparing a diarrhea modeling preparation, comprising the following steps:
[0006] (1) Weigh senna leaves, cascara bark, and atractylodes macrocephala, add water, heat, and perform ultrasonic treatment to obtain a medicinal solution;
[0007] (2) adding β-glucanase, cellulase, phytase, xylanase and glucose oxidase to the drug solution for enzymatic hydrolysis;
[0008] (3) The enzymatically hydrolyzed drug solution is filtered, distilled, and dried to obtain a preparation for diarrhea modeling.
[0009] Specifically, the mass ratio of the above-mentioned senna leaves, sagrada serrata, and atractylodes macrocephala is (30-55): (20-35): (10-50).
[0010] Specifically, in the above step (2), based on the total weight of senna leaves, sagrada bark and atractylodes macrocephala, 5-10% of β-glucanase, 20-35% of cellulase, 3-5% of phytase, 20-45% of xylanase and 3-8% of glucose oxidase are added.
[0011] Specifically, the enzyme activity of the above-mentioned β-glucanase is 10000-20000 U / g, the enzyme activity of cellulase is 5000-10000 U / g, the enzyme activity of phytase is 20000-100000 U / g, the enzyme activity of xylanase is 50000-100000 U / g, the enzyme activity of acid protease is 50000-100000 U / g, and the enzyme activity of glucose oxidase is 4000 U / g.
[0012] Specifically, the enzymatic hydrolysis conditions in the above step (2) are 80-160 rpm, 25-41° C. shaking for 8 h.
[0013] Specifically, the above step (3) further comprises, after drying, crushing the product and mixing it with carboxymethyl cellulose to obtain a preparation for diarrhea modeling.
[0014] The present invention also provides a preparation for diarrhea modeling, which is prepared by the above method.
[0015] The present invention also provides a method for constructing a diarrhea animal model, comprising the following steps:
[0016] (1) preparing the above-mentioned diarrhea modeling preparation;
[0017] (2) The diarrhea modeling preparation is mixed with the basic diet and then fed to animals to obtain a diarrhea animal model.
[0018] Specifically, based on the mass of the basic diet, the amount of the preparation added for diarrhea modeling is 200-500g / t.
[0019] The present invention also provides a diarrhea animal model, which is constructed using the above construction method.
[0020] Compared with the prior art, the present invention has the following advantages and beneficial effects:
[0021] The preparation method of the diarrhea modeling preparation provided by the present invention is to reduce the irritation of the medicinal materials, improve the palatability of the preparation, and avoid the modeling effect affected by constipation of laying hens caused by high-dose senna, so as to prepare a mild, non-toxic, stable and effective laying hen diarrhea agent for the establishment of laying hen diarrhea model, realize efficient and stable modeling, and not easily cause laying hen death. The diarrhea modeling preparation provided by the present invention can be mixed with feed and added, without gavage, reducing the cumbersome operation of traditional senna gavage and esophageal damage caused by manual operation.
[0022] The present invention will be further described in detail below with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1 It is a statistical diagram of diarrhea in laying hens in an embodiment of the present invention.
[0024] Figure 2 It is a statistical diagram of laying hen mortality in an embodiment of the present invention.
[0025] Figure 3 1 is a comparison chart of diarrhea conditions in laying hens in an embodiment of the present invention, wherein: A is a control group, B is a senna leaf group, C is a high salt group, and D is a preparation group. DETAILED DESCRIPTION
[0026] The technical scheme in the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Although the representative embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that various modifications and changes can be made to the present invention without departing from the scope of the present invention. Therefore, the scope of the present invention should not be limited to the embodiments, but should be limited by the attached claims and their equivalents.
[0027] The present invention provides a method for preparing a preparation for diarrhea modeling, comprising the following steps:
[0028] (1) Weigh senna leaves, cascara bark, and atractylodes macrocephala, add water, and heat in a water bath preferably at 60° C. for 4 h. Take out and let stand to room temperature, then use an ultrasonic cell disruptor at 60 W power, 30 s / time, 1 min interval, and repeat ultrasonication for 5 min to obtain a medicinal solution.
[0029] The mass ratio of senna leaf, cascara bark and atractylodes macrocephala is (30-55):(20-35):(10-50).
[0030] (2) Add β-glucanase, cellulase, phytase, xylanase and glucose oxidase to the drug solution and place it in a shaker at 80-160 rpm and 25-41°C for enzymatic hydrolysis for 8 h.
[0031] Based on the total weight of senna leaves, cascara bark and atractylodes macrocephala, the added amount of beta-glucanase is 5-10%, the added amount of cellulase is 20-35%, the added amount of phytase is 3-5%, the added amount of xylanase is 20-45% and the added amount of glucose oxidase is 3-8%.
[0032] Preferably, the enzymatic activity of β-glucanase is 10000-20000 U / g, the enzymatic activity of cellulase is 5000-10000 U / g, the enzymatic activity of phytase is 20000-100000 U / g, the enzymatic activity of xylanase is 50000-100000 U / g, the enzymatic activity of acid protease is 50000-100000 U / g, and the enzymatic activity of glucose oxidase is 4000 U / g.
[0033] (3) filtering and distilling the enzymatically hydrolyzed drug solution until crude crystals are precipitated, and freeze-drying the solution to obtain a crude finished product.
[0034] In order to improve the palatability of the preparation, increase the appetite of livestock and poultry, increase the adhesion of feed particles, and reduce the spillage and waste of raw materials, the crude product is crushed and mixed with carboxymethyl cellulose to obtain a preparation for diarrhea modeling.
[0035] The present invention also provides a method for constructing a diarrhea animal model, comprising the following steps:
[0036] (1) preparing the above-mentioned diarrhea modeling preparation;
[0037] (2) Based on the mass of the basic diet, the diarrhea modeling preparation is added to the basic diet at an addition amount of 200-500 g / t, and then the mixture is fed to the animals to obtain a diarrhea animal model.
[0038] The diarrhea modeling preparation of the present invention, its preparation method and the effects of its application are studied below through specific examples.
[0039] Embodiment 1:
[0040] This embodiment provides a preparation for diarrhea modeling, which is prepared using the following steps.
[0041] (1) Weigh 500 g of dried senna leaves, 300 g of dried bark of Salvia salviae, and 200 g of Atractylodes macrocephala, pick out the rotten and insect-infested parts, add 1000 mL of distilled water, heat in a 60°C water bath for 4 h, take out and let stand to room temperature, use an ultrasonic cell disruptor at 60 W power, 30 s / time, 1 min interval, and repeat ultrasonication for 5 min to obtain a medicinal solution.
[0042] (2) Based on the total mass (1000 g) of senna leaves, sagrada bark, and atractylodes macrocephala, add 8% β-glucanase, 30% cellulase, 4% phytase, 38% xylanase, and 5% glucose oxidase to the medicinal solution, and place it in a shaker at 120 rpm and 37°C for enzymatic hydrolysis for 8 h.
[0043] The enzymatic activity of β-glucanase is 15000U / g, the enzymatic activity of cellulase is 8000U / g, the enzymatic activity of phytase is 60000U / g, the enzymatic activity of xylanase is 70000U / g, the enzymatic activity of acid protease is 70000U / g, and the enzymatic activity of glucose oxidase is 4000U / g.
[0044] (3) The enzymatically hydrolyzed drug solution is placed in a double-layer filter cloth for coarse filtration, the filtrate is poured into a Buchner funnel for suction filtration, the filtrate is distilled under reduced pressure at 70° C. until coarse crystals are precipitated, and freeze-dried to obtain a crude product.
[0045] The crude product was crushed into 40 meshes and mixed with carboxymethyl cellulose at a mass ratio of 1:1 to obtain a preparation for diarrhea modeling.
[0046] Embodiment 2:
[0047] This example studies the effect of the preparation for diarrhea modeling, as follows.
[0048] 1. Experimental Grouping
[0049] 720 healthy and uniformly weighted Hy-Line grey laying hens were selected and randomly divided into 10 treatment groups, with 6 replicates in each group (4 per cage, 3 cages per replicate), and 12 per replicate. The pre-test period was 2 weeks and the main test period was 2 weeks. During the pre-feeding period, the egg production rate between groups was adjusted until the egg production rate between groups was consistent and there was no statistical difference. The age of the laying hens in the formal test was 268 to 282 days old. The test grouping is shown in Table 1.
[0050] Table 1 Test groups
[0051]
[0052] Senna leaves were purchased from Shaanxi Haiyis Biotechnology Co., Ltd., and were mixed with physiological saline. On the day when the formal experiment of laying hens began, the mixture was injected into the crop of laying hens using a rubber tube.
[0053] 2. Experimental diet and feeding management
[0054] The breeding experiment was conducted at the Xinhua Yangtuanfeng Experimental Base from May 14, 2024 to July 8, 2024. The chicken house is a two-layer stacked cage (4 birds / cage, 3 cages / replicate), using a combination of artificial lighting and natural lighting, maintaining 15 hours of light per day (05:00-20:00), light intensity 10-15LX, mechanical ventilation + natural ventilation, room temperature 15-20℃, humidity 40%-60%. Feed twice a day (8:00, 13:00), and the feeding amount is based on basically no leftover feed every day; and the temperature and humidity in the house at these three time points are recorded. Eggs are picked up at 14:00 every day, and they are counted, marked, and stored after picking up. Clean up feces regularly and vaccinate according to routine immunization procedures. During the pre-test, the egg production rate of laying hens was observed, and each group was adjusted so that there was no statistical difference in egg production rate between the groups. Free access to food and drinking water was available throughout the trial.
[0055] The feed was complete crushed feed, and no antibiotics were added to the feed. The composition and nutritional ingredients of the basic diet are shown in Table 2.
[0056] Table 2 Feed composition and nutritional level (air-dried basis)
[0057]
[0058]
[0059] 1) Provide per kg diet: VA 3000 IU, VD 32400 IU, VE 16 IU, VK 32 mg, thiamine (VB1) 1.6 mg, riboflavin (VB2) 6.5 mg, niacin 28 mg, choline chloride 450 mg, calcium pantothenate 9 mg, pyridoxine (VB6) 3.25 mg, biotin 0.07 mg, folic acid 0.8 mg, cyanocobalamin (VB 12 ) 0.015 mg, iron Fe (ferrous sulfate) 267.5 mg, copper Cu (copper sulfate) 13 mg, manganese Mn (manganese sulfate) 95 mg, zinc Zn (zinc sulfate) 90 mg, iodine I 1.35 mg, selenium Se (sodium selenite) 0.29 mg.
[0060] 2)Nutrient levels are calculated values.
[0061] 3. Evaluation indicators
[0062] After the formal experiment of laying hens began, four 5cm×5cm squares were randomly selected from the feces conveying belt of each treatment every two days, and the number of loose feces in each square was counted. Loose feces were scored as 1 point, watery feces were scored as 2 points, and granular feces were scored as -2 points. The diarrhea rate was calculated based on the treatment group. The diarrhea situation of laying hens was as follows: Figure 3 shown.
[0063] The evaluation results of laying hen diarrhea rate are shown in Table 3 and Figure 1shown.
[0064] Table 3 Diarrhea rate index of laying hens during 1-14 days of modeling
[0065]
[0066] The statistical results of laying hen mortality are shown in Table 4 and Figure 2 shown.
[0067] Table 4 Mortality statistics of laying hens from 1 to 14 days after modeling
[0068]
[0069] It can be seen from the experimental data that the modeling using the preparation provided by the present invention has a more stable diarrhea modeling result compared with the gavage feeding of ordinary senna solution, and at the same time, the mortality rate of laying hens is lower than that of modeling with high-salt diets.
[0070] The above examples are merely illustrative of the present invention and do not constitute a limitation on the protection scope of the present invention. All designs that are the same or similar to the present invention fall within the protection scope of the present invention.
Claims
1. A method for preparing a preparation for diarrhea modeling, characterized in that: The following steps are involved: (1) Weigh senna leaves, cascara bark, and atractylodes macrocephala, add water, heat, and perform ultrasonic treatment to obtain a medicinal solution; (2) adding β-glucanase, cellulase, phytase, xylanase and glucose oxidase to the drug solution for enzymatic hydrolysis; (3) The enzymatically hydrolyzed drug solution is filtered, distilled, and dried to obtain a preparation for diarrhea modeling.
2. The method for preparing the diarrhea modeling preparation according to claim 1, characterized in that: The mass ratio of the senna leaf, sagrada serrata and atractylodes macrocephala is (30-55): (20-35): (10-50).
3. The method for preparing the diarrhea modeling preparation according to claim 1, characterized in that: In the step (2), based on the total weight of senna leaves, sagrada bark and atractylodes macrocephala, 5-10% of beta-glucanase, 20-35% of cellulase, 3-5% of phytase, 20-45% of xylanase and 3-8% of glucose oxidase are added.
4. The method for preparing the diarrhea modeling preparation according to claim 3, characterized in that: The enzyme activity of the beta-glucanase is 10000-20000 U / g, the enzyme activity of the cellulase is 5000-10000 U / g, the enzyme activity of the phytase is 20000-100000 U / g, the enzyme activity of the xylanase is 50000-100000 U / g, the enzyme activity of the acid protease is 50000-100000 U / g, and the enzyme activity of the glucose oxidase is 4000 U / g.
5. The method for preparing the diarrhea modeling preparation according to claim 1, characterized in that: The enzymatic hydrolysis conditions in step (2) are 80-160 rpm, 25-41° C. shaking for 8 h.
6. The method for preparing the diarrhea modeling preparation according to claim 1, characterized in that: The step (3) further comprises, after drying, crushing the product and mixing it with carboxymethyl cellulose to obtain a preparation for diarrhea modeling.
7. The preparation for diarrhea modeling prepared by the preparation method according to any one of claims 1 to 6.
8. A method for constructing a diarrhea animal model, characterized in that: The following steps are involved: (1) preparing the diarrhea modeling preparation as claimed in claim 7; (2) The diarrhea modeling preparation is mixed with the basic diet and then fed to animals to obtain a diarrhea animal model.
9. The method for constructing an animal model of diarrhea according to claim 8, characterized in that: Based on the mass of the basic diet, the amount of the preparation added for diarrhea modeling is 200-500g / t.
10. The diarrhea animal model constructed according to the construction method according to any one of claims 8 to 9.