A water-soluble dimethylnidazole composition and its preparation method and application

Abstract

The present invention provides a water-soluble dimethylnidazole composition, a preparation method thereof, and an application thereof, belonging to the technical field of veterinary drug preparation. The composition comprises dimethylnidazole 10-20%, tremella oligosaccharide 2-4%, vitamin K3 2%, hydroxypropyl methylcellulose HPMC E5 0.4-1.2%, xanthan gum 0.1-0.4%, a cosolvent 15-25%, a stabilizer 5-10%, Tween 20 2%, and a water-soluble carrier as the remainder. The composition can reach 8000ppm water-soluble, 12000ppm suspension, uniform dispersion without stratification for 4 hours, and convenient drinking water administration. The combination of dimethylnidazole and tremella oligosaccharide enhances the preventive and therapeutic effect on pathogens such as Escherichia coli and Salmonella, reduces the usage of antibiotics and traditional Chinese medicine extracts, and improves drug bioavailability and efficacy. The composition has a definite therapeutic effect on poultry enteritis and salpingitis caused by such pathogens and is suitable for application in intensive scale farms.

Description

Technical Field

[0001] The invention belongs to the technical field of veterinary drug preparation, and in particular relates to a water-soluble dimethylnidazole composition and a preparation method and application thereof. Background Art

[0002] Dimetridazole is an antiprotozoal drug specifically for animals. Since the banning of metronidazole, dimetridazole has become virtually the only available drug. It inhibits the redox reaction of amoebas, causing nitrogen chain scission. It is effective against protozoa such as Trichomonas, Trichomonas, Balantidium coli, Giardia, and Eperythrozoon. It has a significant inhibitory effect on anaerobic bacteria (Escherichia coli, Streptococcus, Staphylococcus, Necrotic Bacillus, Clostridium, and Vibrio intestinalis). It is also highly effective against spirochetes, which are intermediate between protozoa and bacteria. It is an effective drug for treating swine bloody diarrhea and is commonly used clinically to treat anaerobic infections in animals. It can also be used for diseases such as pseudomembranous enterocolitis caused by Clostridium difficile.

[0003] Dimetridazole's anti-anaerobic effect is primarily due to its ability to spontaneously reduce DNA-bound complexes in anaerobic environments, thereby causing rapid cell death. Dimetridazole's unique, broad-spectrum antimicrobial mechanism of action essentially eliminates the shared or similar antimicrobial targets or mechanisms of action with other known antimicrobial agents, and it also lacks cross-resistance with other antimicrobial agents. Therefore, broad-spectrum antimicrobial agents like dimetridazole are often used in combination with broad-spectrum antimicrobial agents such as amoxicillin, cephalosporins, ofloxacin, colistin sulfate, and sulfonamides in many bacterial and mycoplasma treatment prescriptions to enhance efficacy. However, the long-term and widespread use of antimicrobial agents can lead to the development of strong pathogen resistance, a problem that cannot be ignored. Under the influence of the "green trend", plant-based medicines have attracted much attention for their characteristics of being less likely to develop drug resistance, less likely to remain in livestock products for a long time or to be enriched through the food chain, and being relatively safe for humans, animals and the environment. Therefore, the development of a new, efficient and safe dextran preparation will also be an important research direction for veterinary workers.

[0004] Dimetridazole powder is off-white or slightly yellow, appearing as fibrous crystals or crystalline powder. It is odorless or nearly odorless, has a melting point of 138-139°C, and is slightly soluble in water. Its water solubility is less than 2000 ppm, and its dissolution rate is slow, making it difficult to completely dissolve at room temperature (some insoluble particles require stirring for more than 30 minutes to completely dissolve). For poorly soluble drugs, their bioavailability often depends on the dissolution rate of the solid drug in the formulation. Coarsely dispersed powders prepared by conventional mixing methods often have low bioavailability due to their low dissolution rate. Dimetridazole itself is alkaline, and under alkaline conditions, it has poor stability and low solubility. Dissolution is difficult, it turns black in light, and it easily sublimates at high temperatures. These issues make its utilization rate low and its storage difficult. Therefore, how to improve the stability and water solubility of dimetridazole products is a question worth exploring. Summary of the Invention

[0005] Based on the above problems and in view of the shortcomings and defects of the existing technology of dimetridazole preparations, the present invention provides a water-soluble dimetridazole composition and its preparation method and application, which solves the technical problems of poor water solubility of dimetridazole preparations and precipitation after long-term storage; the combination uses dimetridazole and Tremella oligosaccharides in combination, which enhances the prevention and treatment effect on pathogenic bacteria such as Escherichia coli and Salmonella, and has significant therapeutic effects on enteritis and salpingitis in livestock and poultry caused by such pathogens.

[0006] In order to solve the above technical problems, the technical solution of the present invention is:

[0007] A water-soluble dimenoic acid composition comprises the following components in percentage by weight: dimenoic acid 10-20%, tremella oligosaccharide 2-4%, vitamin K3 2%, hydroxypropyl methylcellulose HPMC E5 0.4-1.2%, xanthan gum 0.1-0.4%, cosolvent 15-25%, stabilizer 5-10%, Tween 20 2%, and the balance a water-soluble carrier.

[0008] According to one embodiment of the present invention, the molecular weight of the Tremella oligosaccharide is 0.6KDa-1.2KDa.

[0009] According to one embodiment of the present invention, the co-solvent is one or more of anhydrous citric acid, tartaric acid, malic acid, succinic acid, and fumaric acid, preferably anhydrous citric acid.

[0010] According to one embodiment of the present invention, the stabilizer is one or more of sodium dihydrogen phosphate, potassium dihydrogen phosphate, sodium hexametaphosphate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, potassium tartrate, and sodium tartrate, preferably potassium dihydrogen phosphate.

[0011] According to one embodiment of the present invention, the water-soluble carrier is one or more of anhydrous glucose, lactose, mannitol, sucrose, sodium chloride, water-soluble starch, and anhydrous sodium sulfate.

[0012] Preferably, the dimenolide composition comprises the following components in weight percentage: dimenolide 20%, tremella oligosaccharide 4%, vitamin K3 2%, hydroxypropyl methylcellulose HPMC E5 0.4%, xanthan gum 0.4%, anhydrous citric acid 25%, potassium dihydrogen phosphate 10%, Tween 20 3% and the balance anhydrous glucose.

[0013] According to another aspect of the present invention, the present invention also provides a method for preparing a water-soluble dimethylnidazole composition, comprising the following steps:

[0014] (1) The above-mentioned amount of dimenoic acid, Tremella oligosaccharide, xanthan gum, and vitamin K3 are micronized by a jet mill to obtain a co-micronized material, wherein the particle size D90 of the co-micronized material is controlled to be 3-10 μm; the other solid materials are all crushed to 80 μm and set aside;

[0015] (2) Dispersing the co-micronized powder, cosolvent, stabilizer, and Tween 20 in warm water to obtain a co-micronized powder solution, wherein the temperature of the warm water is controlled at 40°C-45°C; dissolving Hydroxypropyl Methylcellulose (HPMC) E5 in warm water, and then adding the mixture to the co-micronized powder solution to obtain a material solution, mixing the mixture, and spray drying the mixture;

[0016] (3) The dry powder obtained in step (2) is mixed evenly with a water-soluble carrier to obtain the water-soluble dimethylnidazole composition.

[0017] According to one embodiment of the present invention, the air flow pulverization in step (1) is carried out at a temperature of -3°C to 8°C, an air inlet pressure of 0.9-2.5 MPa, and a working pressure of 0.8-2 MPa during pulverization.

[0018] According to one embodiment of the present invention, the material solution in step (2) has a solid content of 17-23%, and the spray drying conditions are: an inlet air temperature of 140-160°C, an outlet air temperature of 40-60°C, and a feed rate of 10-15 mL / min.

[0019] According to another aspect of the present invention, the present invention also provides use of the above-mentioned dimenoic acid composition in the treatment of enteritis / salpingitis in laying hens.

[0020] Compared with the prior art, the present invention has the following advantages:

[0021] First, the present invention creatively co-powders dimernidazole with tremella oligosaccharides, significantly increasing the water solubility of dimernidazole. Tremella oligosaccharides are then compounded with a small amount of xanthan gum to form a low-viscosity aqueous sol that disperses and encapsulates dimernidazole. Hydroxypropyl methylcellulose HPMC E5 is a linear polysaccharide composed of glucose monomers connected by β-1,4-glycosidic bonds and has a certain water solubility. Its introduction further binds dimernidazole in an interlocking network structure, thereby enhancing the water solubility of the composition, which can reach 8000 ppm. Moreover, when the concentration is too high, the composition sample can be suspended and is stable without stratification when placed for a long time. No additional antioxidant is required, and the composition can be directly added to feed or drinking water as a veterinary drug for consumption. It is easy to use and has a significant therapeutic effect.

[0022] 2. The water-soluble excipients in the existing composition are partially hygroscopic, which easily causes the product to absorb moisture and agglomerate. The addition of Tremella oligosaccharides in the present invention has an unexpected anti-caking effect, and also cooperates with Tween-20 to eliminate static electricity, further improving the dispersibility of the dimethylnidazole ultrafine powder in water.

[0023] 3. The composition of the present invention adds Tremella oligosaccharides of preferred molecular weight, which can further enhance the preventive and therapeutic effects on pathogenic bacteria such as Escherichia coli and Salmonella, and has significant efficacy in treating enteritis and salpingitis in livestock and poultry caused by such pathogenic bacteria. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a diagram of the adhesion of the ultrafine powder device of dimethylnidazole alone.

[0025] Figure 2 This is the adhesion diagram of the equipment after dextromethorphan and Tremella oligosaccharide were co-powdered.

[0026] Figure 3 This is the particle size distribution diagram of dimethylnidazole and Tremella oligosaccharide co-powder.

[0027] Figure 4 This is a comparison picture of dirty eggs and blood-spotted eggs in a chicken farm.

[0028] Figure 5 This is a schematic diagram of the color change of clinical eggs in Example 1 over 5-6 days.

[0029] Figure 6 This is a schematic diagram of the color changes of clinical eggs in Example 1 over 7-9 days.

[0030] Figure 7 A Venn diagram of species distribution.

[0031] Figure 8 Relative abundance map of intestinal microbial sequencing.

[0032] Figure 9 Heatmap of gut microbial functional abundance. DETAILED DESCRIPTION

[0033] To make the purpose and content of the invention of this application more clear, the applicant further describes the present invention with reference to the following embodiments. However, the scope of protection of the present invention is not limited to these embodiments. Those skilled in the art should understand that equivalent substitutions made to the technical features of the present invention, or corresponding improvements, still fall within the scope of protection of the present invention.

[0034] The traditional Chinese medicine extract Tremella oligosaccharide selected in the present invention is a low-polymerized carbohydrate formed by 2 to 10 monosaccharides connected by glycosidic bonds through dehydration condensation, obtained from Tremella through four steps of extraction, degradation, purification and drying. It has a molecular weight of 0.6KDa-1.2KDa. It has a relatively short sugar molecule chain, is soluble in cold water, is easily absorbed by the body, can be used as an energy source, and is well-known for its prebiotic effect, which means that Tremella oligosaccharide can promote the growth of beneficial intestinal bacteria. It has a positive impact on digestive health and immune function. Tremella oligosaccharide itself has good processing characteristics and has the functions of moisturizing, oil absorption, anti-oxidation, thickening and suspending, and emulsification and dispersion.

[0035] The Tremella oligosaccharide used in the present invention can be extracted and separated from Tremella fruiting bodies using conventional extraction and purification methods in the field of natural medicinal chemistry. It is soluble in cold water and can also be purchased from commercial products, for example, Shandong Tongyuan Gonghe Biotechnology Co., Ltd.

[0036] The vitamin K3 in the present invention is mainly used as a hemostatic agent and an intestinal protective agent.

[0037] Unless otherwise specified, the experimental methods used in the following examples and comparative examples are conventional methods; the materials, reagents, etc. used are all commercially available unless otherwise specified.

[0038] Example 1

[0039] A water-soluble dimernidazole composition comprises the following components in percentage by weight: 20% dimernidazole, 4% tremella oligosaccharide, 2% vitamin K3, 0.4% hypromellose HPMC E5, 0.4% xanthan gum, 25% anhydrous citric acid, 10% potassium dihydrogen phosphate, 2% Tween 20, and 36.2% anhydrous glucose.

[0040] The preparation method of the water-soluble dimethylnidazole composition of this embodiment comprises the following process steps:

[0041] (1) The above-mentioned amount of dimenolide, Tremella oligosaccharide, and xanthan gum were micronized by low-temperature airflow grinding (at a temperature of 8° C., an air inlet pressure of 2.5 MPa, and a grinding pressure of 2 MPa) to obtain a co-micronized material with a particle size D90 of 3-10 μm; the other solid materials were all ground to 80 μm and set aside;

[0042] (2) Disperse the co-micronized powder, anhydrous citric acid, potassium dihydrogen phosphate, and Tween 20 in an appropriate amount of warm water at high speed, and control the water temperature at 40°C-45°C; dissolve Hydroxypropyl methylcellulose HPMC E5 in a small amount of warm water, and then add it to the above co-micronized powder solution to obtain a solid content of 17%. After mixing, spray drying is performed, wherein the inlet air temperature is 160°C, the outlet air temperature is 60°C, and the feed rate is 10 mL / min;

[0043] (3) The dry powder obtained in step (2) is evenly mixed with anhydrous glucose to obtain the water-soluble dimenoic acid composition.

[0044] Example 2

[0045] A water-soluble dimenolide composition comprises the following components in percentage by weight: 15% dimenolide, 3% tremella oligosaccharide, 2% vitamin K3, 0.8% hypromellose HPMC E5, 0.25% xanthan gum, 20% tartaric acid, 8% sodium dihydrogen phosphate, 2% Tween 20, and 48.95% sodium chloride.

[0046] The preparation method of the water-soluble dimethylnidazole composition of this embodiment comprises the following process steps:

[0047] (1) The formulated amount of dimethylnidazole, Tremella oligosaccharide, and xanthan gum were micronized by low-temperature airflow grinding (at a temperature of 1° C., an air inlet pressure of 1.5 MPa, and a grinding pressure of 1 MPa) to obtain a co-micronized material with a particle size D90 of 3-10 μm; the other solid materials were all ground to 80 μm and set aside;

[0048] (2) Disperse the co-micronized powder, tartaric acid, sodium dihydrogen phosphate, and Tween 20 at high speed in an appropriate amount of warm water, with the water temperature controlled at 40°C-45°C; dissolve Hydroxypropyl methylcellulose (HPMC) E5 in a small amount of warm water, and then add it to the co-micronized powder solution to a solid content of 19%. Mix well and spray dry. The air inlet temperature is 145°C, the air outlet temperature is 55°C, and the feed rate is 12 mL / min.

[0049] (3) The dry powder obtained in step (2) is mixed evenly with sodium chloride to obtain the water-soluble dimethylnidazole composition.

[0050] Example 3

[0051] A water-soluble dimenolide composition comprises the following components in percentage by weight: 10% dimenolide, 2% tremella oligosaccharide, 2% vitamin K3, 1.2% hypromellose HPMC E5, 0.1% xanthan gum, 15% malic acid, 5% sodium hexametaphosphate, 2% Tween 20, and 62.7% mannitol.

[0052] The preparation method of the water-soluble dimethylnidazole composition of this embodiment comprises the following process steps:

[0053] (1) The formulated amount of dimethylnidazole, Tremella oligosaccharide, and xanthan gum were micronized by low-temperature air flow grinding (at a temperature of -3°C, an air inlet pressure of 0.9 MPa, and a grinding pressure of 0.8 MPa) to obtain a co-micronized material with a particle size D90 of 3-10 μm; the other solid materials were all ground to 80 μm and set aside;

[0054] (2) Disperse the co-micronized powder, malic acid, sodium hexametaphosphate, and Tween 20 at high speed in an appropriate amount of warm water, with the water temperature controlled at 40°C-45°C; dissolve Hydroxypropyl methylcellulose (HPMC) E5 in a small amount of warm water, and then add it to the co-micronized powder solution to obtain a solid content of 23%. Mix well and spray dry the mixture, wherein the inlet air temperature is 140°C, the outlet air temperature is 40°C, and the feed rate is 15 mL / min;

[0055] (3) The dry powder obtained in step (2) is evenly mixed with mannitol to obtain the water-soluble dimethylnidazole composition.

[0056] Comparative Example 1

[0057] A water-soluble dimenoic acid composition comprises the following components in percentage by weight: dimenoic acid 20%, vitamin K3 2%, hydroxypropyl methylcellulose HPMC E5 0.4%, xanthan gum 0.4%, anhydrous citric acid 25%, potassium dihydrogen phosphate 10%, and anhydrous glucose 42.2%.

[0058] Since direct ultrafine grinding of raw materials will cause agglomeration, severe adhesion and poor dispersibility, the preparation method of this comparative example is different from that of Example 1 in that the raw materials are not ultrafine ground, the auxiliary materials are passed through 80 mesh, and then steps (2) and (3) are entered.

[0059] Comparative Example 2

[0060] A water-soluble dimernidazole composition comprises the following components in percentage by weight: 20% dimernidazole, 4% tremella oligosaccharide, 2% vitamin K3, 25% anhydrous citric acid, 10% potassium dihydrogen phosphate, 2% Tween 20, and 37% anhydrous glucose.

[0061] The preparation method is the same as that of Example 1.

[0062] Comparative Example 3

[0063] A water-soluble dimenolide composition comprises the following components in percentage by weight: 20% dimenolide, 4% tremella oligosaccharide, 2% vitamin K3, 0.4% hydroxypropyl methylcellulose (HPMC E5), 0.4% xanthan gum, 2% Tween 20, and 71.2% anhydrous glucose.

[0064] The preparation method is the same as that of Example 1.

[0065] Comparative Example 4

[0066] A water-soluble dimernidazole composition comprises the following components in percentage by weight: 20% dimernidazole, 4% tremella oligosaccharide, 2% vitamin K3, 0.4% hydroxypropyl methylcellulose (HPMC E5), 0.4% xanthan gum, 25% anhydrous citric acid, 2% Tween 20, and 46.2% anhydrous glucose.

[0067] The preparation method is the same as that of Example 1.

[0068] Comparative Example 5

[0069] Commercially available 20% nitroglycerin premix, Inner Mongolia Huatian Pharmaceutical Co., Ltd., batch number 20240110.

[0070] Comparative Example 6

[0071] The difference from Example 1 is that dimenolide is not added to the formula. Other aspects are the same, and the preparation method is also the same.

[0072] Figure 1 This is the adhesion diagram of the ultrafine powder equipment of dimethylnidazole. Figure 2 This is the adhesion diagram of the device after the co-powder of dimethylnidazole and Tremella oligosaccharide. Figure 1 and Figure 2 It is not difficult to see that before co-grinding, some dimethylnidazole is adhered to the lower plate of the grinding chamber, a large amount of it is adhered to the discharge port of the upper plate of the grinding chamber, and a large amount of it is adhered to the upper end of the upper plate of the grinding chamber. After co-grinding, almost no adhesion is seen. Figure 3 This is the particle size distribution diagram of dimethylnidazole and Tremella oligosaccharide co-powder.

[0073] Application Test-Solubility Test

[0074] According to the solubility standard for soluble powders in the 2020 edition of the Chinese Veterinary Pharmacopoeia: Take an appropriate amount of each of the eight samples in Examples 1, 2, 3, and Comparative Examples 1-5, place them in Nessler colorimetric tubes, add water to each tube to make a 100 ml solution of the appropriate concentration (calculated as dimethylnidazole, the concentration is twice the clinical drinking concentration of 100 ppm), turn it upside down 10 times at 23-27 ° C, and the test sample should be completely dissolved. Let it stand for 30 minutes without turbidity or precipitation. The results showed that all eight samples were water-soluble.

[0075] At room temperature (10-30°C), weigh an appropriate amount of 7 samples from Examples 1, 2, 3 and Comparative Examples 1, 2, 3 and 4, respectively, add water to make 100 ml of solution, and stir for 30 minutes. During the stirring process, check the dissolution of the sample and observe the maximum solubility. The results are shown in Table 1 below.

[0076] Table 1: Dissolution of each test sample

[0077]

[0078] If it dissolves quickly within 5 minutes, mark it with “√”, otherwise mark it with “×”

[0079] Test Conclusion: All the examples were water-soluble at 8000 ppm, differing only in the time it took to dissolve in water. However, none of the comparative examples were water-soluble under these conditions. This suggests that the ultrafine grinding, water-soluble reticulated gel, cosolvent, and dispersant enhance the water solubility and dissolution rate of dimetridazole. Comparative Example 5 is a common product available on the market, which can only achieve 500ppm water solubility within 5 minutes. After dissolving in water, obvious dimernidazole crystal particles can be seen. In about 30 minutes, a concentration of 1000ppm can be slowly dissolved; Comparative Example 1 does not add tremella oligosaccharides and Tween 20. After dissolving in water, small hard granular substances are found, with poor dispersibility, and the water solubility is slightly better than Comparative Example 5; Comparative Example 3 does not add cosolvents and stabilizers, which reduces the water solubility of dimernidazole; Comparative Example 2 does not add xanthan gum and hydroxypropyl methylcellulose HPMC E5, indicating that the water-soluble network glue formed by tremella oligosaccharides, xanthan gum and hydroxypropyl methylcellulose does improve the solubility of dimernidazole. Comparative Example 4 does not add stabilizers, and unstable precipitation occurs after the sample is dissolved in water.

[0080] Application Test 2: Suspension and Homogeneity Test

[0081] The dimenolide composition examples 1-3 prepared by the present invention and the commercially available sample 5 were prepared into a 12000 ppm suspension. After 4 hours, the stratification of the sample was observed, and samples were slowly taken from the top, middle, and bottom of the solution. The sample was diluted 50 times and the dimenolide content was determined according to the dimenolide content method of the 2020 edition of the Veterinary Pharmacopoeia of the People's Republic of China. The results are shown in Table 2.

[0082] Table 2 Results of content determination at different positions of suspension

[0083] Sampling location Concentration 1 (μg / mL) Concentration 2 (μg / mL) Mean concentration (μg / mL) superior 234.12 235.38 234.75 Example 1 middle 237.27 238.46 237.87 Down 241.73 241.54 241.64 superior 235.25 235.41 235.33 Example 2 middle 238.67 239.12 238.90 Down 243.32 242.83 243.08 superior 229.16 230.42 229.79 Example 3 middle 235.24 234.96 235.10 Down 240.18 239.94 240.06

[0084] As can be seen from the results in the table, after standing for 4 hours, samples were taken from the top, middle and bottom of the suspension. The concentration gradually increased from top to bottom, and the difference in the overall content was not obvious. This shows that the samples prepared by the present invention can basically achieve stable suspension within 4 hours at a concentration of 12000ppm. Comparative Example 5 completely sank to the bottom within 4 hours, and the upper solution was clear, so no samples were taken for measurement.

[0085] Application Test 3 Stability Test

[0086] The products of Examples 1-3 and Comparative Examples 1-4 were subjected to high-temperature stability tests and accelerated tests to examine the stability of the samples of the present invention, confirm whether discoloration, caking, deliquescence, etc. occurred, whether the content was stable, and to test whether their water solubility was qualified. The water solubility test method was the same as that of Application Test 1.

[0087] 1. High temperature stability test

[0088] The samples were placed in an open container at 60°C for 10 days, then returned to room temperature for observation. The results showed that the appearance of Examples 1-3 and Comparative Examples 2-4 did not change significantly, with no discoloration or caking. The water solubility and content did not change significantly compared to the initial values, indicating that Examples 1-3 and Comparative Examples 2-4 had good stability at high temperatures. Comparative Example 1 exhibited high-temperature caking, indicating that the sample was prone to high-temperature caking without the protection of Tremella oligosaccharides.

[0089] 2. Accelerated testing

[0090] The samples were packaged for listing and placed in an accelerating box at a temperature of 40°C and a relative humidity of 75% for 6 months, and were sampled and observed every 30 days. The results showed that no significant changes occurred in the appearance of the samples of Examples 1-3 and Comparative Examples 2-4, such as no discoloration, caking, or deliquescence, and no significant changes were observed in the water-soluble and content values ​​compared with the initial values, indicating that the sample stability of Examples 1-3 and Comparative Examples 2-4 was good. The hygroscopic caking phenomenon occurred in Comparative Example 1, indicating that the sample easily absorbed moisture and agglomerated under the moisture-proof protection of Tremella fuciformis oligosaccharides.

[0091] Application Test IV Clinical Efficacy Trial

[0092] Observation on the therapeutic effect of enteritis / salpingitis in laying hens

[0093] Location: A laying hen breeding company in Gansu

[0094] Test subjects: approximately 48,014 laying hens aged 320 days. Recently, the number of dirty eggs, broken eggs, and white-shelled eggs in the flock increased, and the feces were poorly formed. Autopsy showed inflammation of the intestines and bleeding spots in the oviducts. Complications of enteritis and salpingitis caused by infections such as Escherichia coli and Salmonella were diagnosed.

[0095] Trial groups:

[0096] 1) Blank group: no drug was used, 8,000 laying hens;

[0097] 2) Drug control group 1: using the sample of Comparative Example 5, 8005 laying hens;

[0098] Drug control group 2: using the sample of comparative example 6, 8,000 laying hens;

[0099] 3) Experiments 1-3: Using the samples of Examples 1-3, the number of laying hens was 8002, 8004, and 8003, respectively;

[0100] Experimental method: The experimental groups 1 to 3 and the drug control group were all administrated through drinking water. The drinking water concentration was calculated as 100 ppm based on the total daily drinking water volume. Each group was administrated for 5 consecutive days. The group was observed for 10 consecutive days from the start of administration, and the drinking water, feed intake, feces, egg production rate, qualified egg rate, mortality rate and other indicators of the laying hens were recorded.

[0101] (1) Water intake statistics

[0102] The statistics of water drinking during the treatment period showed no significant difference among the groups (P>0.05), which proved that the drugs in each group would not affect the palatability of the chickens.

[0103] (2) Feed intake statistics

[0104] There was no significant difference in feed intake between the groups during the treatment period (P>0.05), which proved that the drugs in each group had no effect on feed intake.

[0105] (3) Stool condition

[0106] Before medication (Day 1), the overall feces formation of the farm was poor, with a proportion of loose feces of about 50%. At the same time, obvious feed feces were visible, about 20%. Observation on the third day of medication (Day 4) showed that each experimental group had significant improvement compared with the blank group, with loose feces and feed feces decreasing by nearly 15% respectively, especially in Experimental Groups 1-3, where loose feces were controlled at 25%, which was better than the other experimental groups. On the fifth day of medication (Day 6), the proportion of abnormal feces in Experimental Groups 1-3 was controlled at around 20%, which was better than the 35% of Comparative Example 5, and the feces improved significantly. Continued follow-up to the fifth day after medication (Day 11), in Experimental Groups 1-3, loose feces were maintained at 5%, and feed feces were controlled at 5%. The loose feces of the comparative group were maintained at 10%, and feed feces were controlled at 10%. This shows that Examples 1-3 ensure effective regulation of the intestinal tract of the chickens and have good prevention and control effects. See Table 3 below for details:

[0107] Table 3 Fecal status of each experimental group

[0108]

[0109] (4) Mortality rate

[0110] During the trial, the number of deaths in each group was counted daily, and the results are shown in Table 4. Each group experienced varying degrees of death every day. The total death rate over 10 days showed that the number of deaths in the experimental groups was reduced compared to the blank group. In particular, the mortality rates of Examples 1-3 were reduced by 75%, 63%, and 63%, respectively. Comparative Example 6 was reduced by 12%, and Comparative Example 5 was reduced by 25%. The Example group showed a highly significant reduction and was superior to the other experimental groups.

[0111] Table 4 Death rate of each experimental group

[0112] Group Mortality rate (‰) Blank group 1 Comparative Example 6 0.88 Comparative Example 5 0.75 Example 1 0.25 Example 2 0.37 Example 3 0.37

[0113] (5) Egg production

[0114] During the test period, the good egg rate of each group was statistically calculated, and the results are shown in Table 5. The egg production rate results after 10 days showed that the test group and the comparative group were improved compared with the blank group, especially the groups of Examples 1-3, where the good egg rate increased from 75% to about 90%, the dirty egg rate decreased from about 7.5% to about 2%, and the blood egg rate decreased from 6% to about 1.5%. There was a significant improvement in egg production. Among them, the comparison of dirty eggs and blood-spotted eggs in the blank group and the group of Example 1 is shown in Table 5. Figure 4 .

[0115] In addition, during the test, it was found that the color of the eggs in the blank group and the comparative example 6 group was lighter, with some white shell eggs present. The eggshell color of the comparative example 5 group improved, but was worse than that of the example group. Figure 5 , 7 to 9 days eggshell changes see Figure 6 .

[0116] Table 5 Egg production of each experimental group

[0117]

[0118]

[0119] (6) Impact on intestinal microbial diversity

[0120] Since the blank group and comparative example 6 were similar in the above test conditions, 50 g of feces were selectively collected from the chicken farms of comparative example 6 (KB2), comparative example 5 (KB1), embodiment 1 (STK1), and implementation 2 (STK2) for 16S-rDNA sequencing. The species distribution Venn diagram is shown in Figure 7 , relative abundance diagram is shown in Figure 8 , see 9 for the function heat map.

[0121] Figure 7 It can be seen that the STK group of Example completely covers the KB control group of Comparative Example, and has 272 more species, which is more species diverse.

[0122] Figure 8 In the relative abundance graph, the abundance of beneficial bacteria in the STK group of Example 1 increased, which is more conducive to maintaining intestinal health. The control group of Comparative Example 6 has more Proteobacteria, mainly pathogenic bacteria. This shows that the addition of a small amount of Tremella oligosaccharides has little effect on the intestine. The proportion of Proteobacteria in Comparative Example 5 decreased by about 20% compared with the Comparative Example 6 group, indicating that nitroglycerin still has an effect on pathogenic bacteria. The proportion of Proteobacteria in the STK group decreased significantly, falling below 10%, and that in Example 1 it dropped to about 5%. This shows that the combination of a small amount of Tremella oligosaccharides and nitroglycerin increases the effect of the product on Proteobacteria and enhances the abundance of intestinal microorganisms in laying hens.

[0123] Proteobacteria is the largest phylum of bacteria, including many pathogens, such as Escherichia coli, Salmonella, Vibrio cholerae, Helicobacter pylori and other well-known species.

[0124] Firmicutes (Firmicutes) primarily break down complex nutrients such as cellulose, starch, and protein, while producing beneficial metabolites. Many members are beneficial bacteria, such as Lactobacillus, Bacillus, Eubacterium, and Roseburia.

[0125] Bacteroidetes (Bacteroidetes) primarily produce enzymes to break down food, aiding in the digestion of nutrients such as carbohydrates, proteins, and fats. They can accelerate angiogenesis in the intestinal mucosa, enhance host immunity, and maintain a balanced intestinal microbiome.

[0126] Fusobacteria are a small group of Gram-negative bacteria that produce beneficial metabolites such as pyruvate and butyrate. Fusobacteria may be involved in maintaining gut microbiota homeostasis and metabolic function.

[0127] Figure 9 In the function heat map, color represents the abundance of the function: redder colors indicate higher abundance, and greener colors indicate lower abundance. The results showed that the STK group had darker colors than the KB group, with STK1 being the darkest and KB1 being the greenest. This suggests that the STK group had increased abundance of beneficial bacteria.

[0128] In summary, the effects of Examples 1-3 are significant in clinical application for enteritis and salpingitis caused by infections such as Escherichia coli and Salmonella, increase the abundance of beneficial intestinal microbial flora, improve loose stools and feed stools caused by diarrhea; reduce the mortality rate; increase the egg production rate, and reduce dirty eggs and blood-stained eggs.

[0129] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention, which are also considered to be within the scope of protection of the present invention.

Claims

1. A water-soluble dimethylnidazole composition, characterized in that The invention comprises the following components in percentage by weight: 10-20% of dimenolide, 2-4% of tremella oligosaccharide, 2% of vitamin K3, 0.4-1.2% of hydroxypropyl methylcellulose (HPMC) E5, 0.1-0.4% of xanthan gum, 15-25% of cosolvent, 5-10% of stabilizer, 2% of Tween 20, and the balance of a water-soluble carrier. The Tremella oligosaccharide is a low-polymer sugar obtained by extracting, degrading, purifying and drying Tremella, which is formed by dehydration condensation of 2 to 10 monosaccharides connected by glycosidic bonds and has a molecular weight of 0.6KDa-1.2KDa.

2. A water-soluble dimethylnidazole composition according to claim 1, characterized in that The cosolvent is one or more of anhydrous citric acid, tartaric acid, malic acid, succinic acid, and fumaric acid.

3. A water-soluble dimethylnidazole composition according to claim 2, characterized in that, The cosolvent is anhydrous citric acid.

4. A water-soluble dimethylnidazole composition according to claim 1, characterized in that The stabilizer is one or more of sodium dihydrogen phosphate, potassium dihydrogen phosphate, sodium hexametaphosphate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, potassium tartrate, and sodium tartrate.

5. A water-soluble dimethylnidazole composition according to claim 4, characterized in that, The stabilizer is potassium dihydrogen phosphate.

6. A water-soluble dimenoic acid composition according to claim 1, characterized in that The water-soluble carrier is one or more of anhydrous glucose, lactose, mannitol, sucrose, sodium chloride, water-soluble starch, and anhydrous sodium sulfate.

7. A water-soluble dimethylnidazole composition according to claim 6, characterized in that The dimenolide composition comprises the following components in percentage by weight: dimenolide 20%, tremella oligosaccharide 4%, vitamin K3 2%, hydroxypropyl methylcellulose HPMC E5 0.4%, xanthan gum 0.4%, anhydrous citric acid 25%, potassium dihydrogen phosphate 10%, Tween 20 2%, and the balance anhydrous glucose.

8. A method for preparing the water-soluble dimethylnidazole composition according to any one of claims 1 to 7, characterized in that: The following steps are involved: (1) The above-mentioned amount of dimenoic acid, Tremella oligosaccharide, xanthan gum, and vitamin K3 are micronized by a jet mill to obtain a co-micronized material, wherein the particle size D90 of the co-micronized material is controlled at 3-10 μm; the other solid materials are all crushed to 80 μm and set aside; (2) Disperse the co-micronized powder, cosolvent, stabilizer, and Tween 20 in warm water, the temperature of which is controlled at 40°C-45°C; dissolve Hydroxypropyl methylcellulose (HPMC) E5 in warm water, and then add it to the co-micronized powder solution to obtain a material solution, mix well, and then spray dry; (3) The dry powder obtained in step (2) is mixed evenly with a water-soluble carrier to obtain the water-soluble dimethylnidazole composition.

9. The preparation method according to claim 8, characterized in that The air flow pulverization in step (1) is carried out at a temperature of -3°C to 8°C, an air inlet pressure of 0.9-2.5 MPa, and a working pressure of 0.8-2 MPa during pulverization.

10. The preparation method according to claim 8, characterized in that The solid content of the material solution in step (2) is 17-23%, and the spray drying conditions are: inlet air temperature is 140-160°C, outlet air temperature is 40-60°C, and feed rate is 10-15 mL / min.

11. Use of the dimenoic acid composition according to any one of claims 1 to 7 in preparing a medicament for treating enteritis / salpingitis in laying hens.

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