Traditional Chinese medicine oral liquid preparation for treating diarrhea syndrome of newborn livestock and preparation method of traditional Chinese medicine oral liquid preparation

Abstract

The invention relates to the technical field of veterinary drugs, and particularly discloses a traditional Chinese medicine oral liquid preparation for treating diarrhea syndrome of newborn livestock and a preparation method thereof. The traditional Chinese medicine oral liquid preparation takes stenoloma chusanum, Chinese ilex leaves and liquorice as raw materials to prepare traditional Chinese medicine liquid, and then functional bacterial powder is added and mixed to prepare the oral liquid. According to the preparation, through the synergistic effect of traditional Chinese medicines and bacterial powder, core causes such as pathogenic bacterium infection, intestinal flora imbalance, mucosal lesion and low immunity of newborn livestock and poultry diarrhea are precisely targeted, rapid diarrhea checking is achieved, the cure rate is remarkably increased, and the recurrence rate is reduced; meanwhile, intestinal barriers are repaired, the body resistance is enhanced, nutrient absorption and post-disease growth are promoted, residues are avoided, drug resistance is avoided, the feed is adaptive to physiological characteristics of newborn livestock and poultry, and the resistance reducing requirement of green breeding is met.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of veterinary drugs, and particularly relates to a traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome and a preparation method thereof. BACKGROUND

[0002] Neonatal diarrhea syndrome is a common disease with complex causes. Various neonatal animals such as calves, foals, piglets, lambs and camel lambs can all occur, and the disease is characterized by acute diarrhea, progressive dehydration and fast death. The disease most commonly occurs in 2-10-day-old neonatal animals, and can occur as early as 2-18 hours after birth, and occasionally as late as 3 weeks of age. The earlier the onset, the higher the mortality rate.

[0003] The disease has complex causes, and biological factors, immune levels of neonatal animals, dietary properties, management levels and mutual reactions between climate factors can all lead to the disease.

[0004] Biological factors: different serotypes of Escherichia coli, certain Salmonella, Clostridium perfringens A, B, C and E, rotavirus and coccidia can cause various enteritis and diarrhea in neonatal animals; and the causes of diarrhea syndrome are different for different types of neonatal animals, and infectious rhinotracheitis virus and coronavirus are easy to cause ruminant diarrhea, chlamydia and cryptosporidium are easy to cause calf diarrhea, and infectious gastroenteritis virus is easy to cause piglet diarrhea.

[0005] Immune level of neonatal animals: the immune ability of young animals is very low in the first few days after birth, and there is no antibody in the body at birth, so neonatal animals must rely on colostrum intake within a few hours after birth to obtain antibodies and other nutrients to improve disease resistance. If the neonatal animals eat colostrum too late, not enough or cannot eat colostrum, the disease resistance will decrease.

[0006] Management and hygiene: deterioration of milk products or poor hygiene, non-sterilization of milk utensils; uneven feeding of young animals leading to dietary diarrhea; unclean, non-periodic sterilization of the delivery room and neonatal animal pens; poor ventilation, overcrowding, lack of cold, wind, heat and rain protection facilities and other factors can all induce the disease.

[0007] Climate factors: the incidence of diarrhea increases significantly in rainy seasons, cold and windy seasons and hot and humid seasons.

[0008] The pathological occurrence of the disease is mainly: ① the invasion of biological factors destroys the intestinal epithelial cells and causes inflammation; ② the increase of osmotic pressure in the intestinal lumen increases the water content in the intestinal lumen; ③ the increase of intestinal secretion caused by bacterial toxins and other reasons; ④ the shortening of the residence time of intestinal contents in the intestinal tract due to the enhancement of intestinal peristalsis, which reduces the ability of the large intestine to absorb water. Under the action of the above factors, neonatal animals suffer from diarrhea, dehydration, acidosis, circulatory failure, shock and death.

[0009] The main symptoms of this disease are diarrhea, dehydration (sunken eyes, decreased skin elasticity, thickened blood, slow venous blood flow, and reduced blood volume), and weakness, with death occurring within one to several days after onset. Symptoms may vary slightly depending on the cause. Diarrhea in newborn animals is mostly caused by Escherichia coli, with onset typically within 5 days of age. It has an acute course, with healthy and active young animals becoming weak after 12-24 hours. Initially, the animals experience abdominal distension and watery, yellowish-white, or green feces. Salmonella infection primarily affects animals older than 2 weeks, with foul-smelling feces often containing blood and large amounts of mucus. Clostridium species cause hemorrhagic enterotoxemia, characterized by sudden depression and weakness, watery diarrhea, and the rapid addition of fibrin, necrotic tissue fragments, and blood to the feces, leading to death within hours. Neurological symptoms are occasionally observed. Rotavirus causes diarrhea in 1-2 week old calves. If not complicated by Escherichia coli infection, the watery diarrhea lasts for several hours and resolves within 18-24 hours. Coronavirus infection in calves aged a few days to a few weeks presents with symptoms similar to rotavirus. Dietary diarrhea initially presents with large amounts of slightly thick, pasty feces, which worsen into thin, watery stools.

[0010] A preliminary diagnosis can be made based on the age of onset and clinical symptoms. A definitive diagnosis requires the culture and isolation of the pathogen and examination for parasites. Fresh fecal samples from surviving infected animals should be taken for testing. However, this is challenging because diarrhea caused by different pathogens is not significantly different and is often a mixed infection. Furthermore, the pathogens can be isolated from healthy young animals, so careful diagnosis is essential.

[0011] Currently, this disease is widespread worldwide, causing enormous economic losses to newborn livestock, and its occurrence and spread are becoming increasingly serious. Sick and carrier animals are the main sources of infection. Young animals with pathological changes but no clinical symptoms, or carrier animals with negative results but no clinical symptoms or pathological changes, are more common. If secondary or concurrent diseases occur, it often leads to aggravation of clinical symptoms and an increase in mortality.

[0012] In recent years, neonatal diarrhea syndrome has become a significant disease affecting the development of animal husbandry. Its incidence in my country has been increasing year by year, making it one of the most serious diseases affecting young livestock and causing severe economic losses. Timely treatment with antibiotics after onset can significantly reduce animal mortality. Appropriate addition of antibiotics during production can also reduce the incidence to some extent. However, with the widespread use of numerous antibiotics, especially broad-spectrum antibiotics, bacterial resistance to antimicrobial drugs is becoming increasingly prominent. Meanwhile, research data also indicates that sub-inhibitory concentrations of antibiotics can affect bacterial metabolic processes, altering bacterial virulence or their adaptability to the environment.

[0013] Early treatment with antibiotics is effective in reducing mortality. However, intramuscular or subcutaneous injections typically require repeated administration of large doses, which can easily lead to drug resistance and serious drug residues, affecting food safety. To effectively control neonatal diarrhea syndrome and address the potential antibiotic residue risks associated with antibiotic prevention and treatment, it is necessary to provide a healthier, residue-free formulation for controlling neonatal diarrhea syndrome. Summary of the Invention

[0014] In view of the shortcomings and problems of antibiotic residues in the current treatment of neonatal diarrhea syndrome, the present invention provides a traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome and its preparation method.

[0015] This invention provides a traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome, comprising the following raw materials in parts by weight: 3-6 parts of fern, 3-6 parts of holly leaves, 5-10 parts of licorice, 1-3 parts of sodium chloride, 1-2 parts of potassium chloride, 1-3 parts of sodium bicarbonate, 1-8 parts of glucose, and 1-6 parts of glycine.

[0016] The above-mentioned oral liquid preparation of traditional Chinese medicine for treating neonatal diarrhea syndrome includes the following raw materials in the following weight ratios: 6 parts of fern, 6 parts of holly leaf, 10 parts of licorice, 3 parts of sodium chloride, 2 parts of potassium chloride, 3 parts of sodium bicarbonate, 8 parts of glucose, and 6 parts of glycine.

[0017] The aforementioned oral liquid preparations of traditional Chinese medicine used to treat neonatal diarrhea syndrome also include Bacillus saffron and Bacillus urealyticum powder.

[0018] This invention also provides a method for preparing a traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome, comprising the following steps: I. Preparation of Traditional Chinese Medicine Liquid: (1) Crush the fern into 50 mesh, put it into an extraction tank, add 6 times the amount of 80% ethanol and soak for 1 hour, then boil at 60℃ for 30 minutes, extract the supernatant, concentrate under reduced pressure to recover the ethanol, boil into an extract, add sterile ultrapure water to the extract, filter out impurities, and make it into an aqueous solution with a concentration of 1:1~1.2. (2) Crush the holly leaves to 50 mesh, put them into an extraction tank, add 5 times the amount of water and soak for 1 hour, then decoct at 75°C for 1.5 hours and extract the supernatant; add 5 times the amount of water a second time, decoct at 80°C for 2 hours and extract the supernatant; combine the supernatants and concentrate to 1 / 3 volume. (3) Add licorice to the extraction tank, add 5 times the amount of water and soak for 1 hour, then decoct at 100°C for 30 minutes and extract the supernatant; add 5 times the amount of water again, decoct at 80°C for 2 hours and extract the supernatant; combine the supernatants and concentrate to 1 / 3 volume. (4) Combine the three liquids, add sterile ultrapure water, filter through a microporous membrane to remove impurities, and make the concentration of the traditional Chinese medicine solution 1:1~1.2; II. Preparation of mycelium powder: (1) Inoculate Bacillus saffron and Bacillus urealyticum onto tryptic soybean agar solid medium and incubate at 28-30 °C for 24-48 hours; (2) Take the strain cultured in step (1) and inoculate it into tryptic soybean liquid culture medium. Culture it at 28-30 ℃ for 24-48 h to obtain seed liquid; (3) Inoculate the seed liquid into the fermentation medium at an inoculation rate of 2-3%, and ferment and culture it for 24-48 hours at 28-30 ℃, 80-120 rpm and pH 6.8-7.2. Collect the fermentation liquid to obtain the microbial agent; spray the liquid into powder using a powder spraying device.

[0019] III. Preparation of oral liquid formulations: The traditional Chinese medicine liquid is diluted and sterilized according to the specified ratio and then cooled naturally. 0.35% sodium chloride, 0.1% potassium chloride, 0.3% sodium bicarbonate, 1% glucose, and 0.5% glycine are dissolved in sterile water and then aseptically mixed with the sterilized traditional Chinese medicine liquid. After that, bacterial powder is added and mixed well to obtain the oral liquid preparation.

[0020] In the preparation method of the above-mentioned oral liquid preparation of traditional Chinese medicine for treating neonatal diarrhea syndrome, the dilution ratio of the traditional Chinese medicine liquid in step three is 25%-50%.

[0021] The above-mentioned method for preparing a traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome, wherein the bacterial count of Bacillus sabinatus in the oral liquid preparation is not less than 10. 6 cfu / mL.

[0022] This formula has the effects of regulating qi and relieving pain, harmonizing the stomach and stopping diarrhea, relieving stagnation of qi in the stomach and intestines, indigestion, loss of appetite, abdominal distension and pain, regulating qi and strengthening the spleen, drying dampness and resolving phlegm, soothing the liver and stopping stagnation, breaking up qi and eliminating stagnation, clearing heat and drying dampness, purging fire and detoxifying, cooling blood and stopping dysentery, regulating the gastrointestinal tract, and replenishing and balancing body fluids.

[0023] Black fern, slightly bitter, and cold in nature; it has the effects of clearing heat and dampness, detoxifying and stopping bleeding; Holly leaves have the effects of clearing heat and detoxifying, promoting tissue regeneration and wound healing, and promoting blood circulation and stopping bleeding;

[0024] Licorice is sweet and neutral in nature. It enters all twelve meridians, replenishes the middle energizer and invigorates qi, clears heat and detoxifies, and moderates the properties of other medicines. It can also reduce the mild intestinal stress response caused by the initial colonization of the intestinal tract by the bacterial powder, reduce the stimulation of the intestine by the Chinese medicine components, and improve the palatability of the preparation.

[0025] The phenolic acid and glycyrrhizic acid in the herbal decoction can directly repair damaged intestinal mucosa and form a protective film. Meanwhile, after colonization, the bacterial powder metabolizes and produces short-chain fatty acids, lowering the intestinal pH and providing a suitable environment for the repair of mucosal epithelial cells. Simultaneously, it inhibits the adhesion and damage of harmful bacteria to the mucosa. Together, these two processes repair damaged mucosa and maintain mucosal homeostasis, rapidly restoring intestinal barrier function and reducing the recurrence of diarrhea. Furthermore, the esterases and glycosidases secreted by the bacterial powder can degrade macromolecular components in the herbal decoction, improving the bioavailability of active ingredients while enhancing their anti-inflammatory, antibacterial, and immunomodulatory activities, thus amplifying the therapeutic effects of the herbal medicine.

[0026] To verify the effect of the prepared traditional Chinese medicine solution on the activity of the bacterial powder, the plate count method was used for activity determination, specifically: The herbal liquid filtered and purified in step one was diluted with 0.85% (w / v) sterile physiological saline to produce 100% stock solution, 75% diluted solution, 50% diluted solution, and 25% diluted solution, and then sterilized. Take several sterile test tubes and add 9 mL of sterile Chinese medicine solution of different concentrations to each as the test group. At the same time, set up a positive control group (add 9 mL of sterile broth) and a negative control group (add 9 mL of 100% sterile Chinese medicine solution, without inoculation). After activating the Bacillus urealyticus powder prepared in step two, 1 ml of Bacillus urealyticus bacterial solution was precisely added to each test tube of the experimental group and the positive control group on a clean bench to obtain a mixed solution. Then, at 0h, 2h, 6h, 12h, and 24h, each test tube of the experimental group and the positive control group was serially diluted to 10⁻¹⁰. -6 ; Take 100ml of the mixture and spread it evenly on the surface of a nutrient agar plate using a sterile spreader; After preliminary experiments, select 10 -2 10 -3 10 -4 Further testing was conducted on the dilutions, with three replicates for each dilution; all plates were inverted and placed in an incubator at 37°C for 24 hours.

[0027] After the experiment, plates with colony counts between 30 and 300 were selected for counting and the viable cell concentration was calculated. A curve was plotted with time on the x-axis and the logarithm of viable cell count on the y-axis to observe the growth status of strains in different experimental groups.

[0028] The same method was used for Bacillus saforticus. The results are shown in Table 1 below.

[0029]

[0030] The results showed that no bacteria grew in the negative control group, and the overall number of viable bacteria in the positive control group did not change significantly, indicating that the bacterial powder prepared by this invention had good activity. However, the 100% stock solution and 75% dilution had a strong immediate inhibitory effect on Bacillus urealyticum and Bacillus safranin, and the 24-hour survival rate was low, so they were not suitable as the concentration of the oral liquid. Considering the 24-hour survival rate and the number of viable bacteria, the traditional Chinese medicine liquid with a dilution of 50% and 25% can be selected for the preparation of the oral liquid.

[0031] To further verify the viable bacterial count of the 50% and 25% diluted traditional Chinese medicine solutions during long-term storage, the oral liquid products made from the 50% and 25% diluted solutions were refrigerated at 4°C, and samples were taken on days 7, 14, 30, and 60 for viable bacterial count determination. The results showed that the total viable bacterial count of both solutions remained above 10 throughout the 60-day period. 6 CFU / mL, meeting the bacterial count requirements for oral liquids.

[0032] Based on the above, a 50% dilution of the drug solution was ultimately chosen for the preparation of the oral liquid to ensure both efficacy and viable bacterial count.

[0033] Compared with existing technologies, the beneficial effects of this invention are as follows: By strictly controlling the processing methods of *Aconitum carmichaelii*, *Ilex chinensis* leaves, and licorice, and by performing microfiltration, dilution, and pH adjustment on the medicinal liquid, this invention effectively reduces the impact of the traditional Chinese medicine liquid on the activity of the bacterial powder, allowing the bacterial powder in the prepared oral liquid to work synergistically with the traditional Chinese medicine liquid. Through the scientific combination of traditional Chinese medicine liquid and bacterial powder, this invention synergistically regulates the balance of intestinal flora and enhances immunity. It not only cures current diarrhea but also repairs intestinal function, improves digestive and absorption efficiency, and reduces diarrhea recurrence. Simultaneously, it promotes weight gain in newborn livestock and poultry after illness, avoiding growth retardation and poor development caused by diarrhea. It achieves multi-target synergistic treatment of antibacterial and bactericidal effects, mucosal repair, flora regulation, immune enhancement, and digestive improvement, precisely addressing the core causes of newborn livestock and poultry diarrhea syndrome. Compared with single traditional Chinese medicine preparations or single bacterial powders, this invention has significant advantages such as rapid diarrhea relief, high cure rate, low recurrence rate, good safety, and growth promotion. Detailed Implementation

[0034] To enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail below with reference to specific embodiments.

[0035] Example 1: This example provides a traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome, comprising the following raw materials in the indicated weight ratios: 6 parts of *Pteris vittata*, 6 parts of *Ilex chinensis* leaves, 10 parts of licorice root, 20 parts of *Bacillus sphaeroides*, 16 parts of *Bacillus urealyticum*, 3 parts of sodium chloride, 2 parts of potassium chloride, 3 parts of sodium bicarbonate, 8 parts of glucose, and 6 parts of glycine; the preparation method includes the following steps: I. Preparation of Traditional Chinese Medicine Liquid: (1) Crush the fern into 50 mesh, put it into an extraction tank, add 6 times the amount of 80% ethanol and soak for 1 hour, then boil at 60℃ for 30 minutes, extract the supernatant, concentrate under reduced pressure to recover the ethanol, boil into an extract, add sterile ultrapure water to the extract, filter out impurities, and make it into an aqueous solution with a concentration of 1:1~1.2. (2) Crush the holly leaves to 50 mesh, put them into an extraction tank, add 5 times the amount of water and soak for 1 hour, then decoct at 75°C for 1.5 hours and extract the supernatant; add 5 times the amount of water a second time, decoct at 80°C for 2 hours and extract the supernatant; combine the supernatants and concentrate to 1 / 3 volume. (3) Add licorice to the extraction tank, add 5 times the amount of water and soak for 1 hour, then decoct at 100°C for 30 minutes and extract the supernatant; add 5 times the amount of water again, decoct at 80°C for 2 hours and extract the supernatant; combine the supernatants and concentrate to 1 / 3 volume. (4) Combine the three liquids, add sterile ultrapure water, filter to remove impurities, and make the concentration of the traditional Chinese medicine solution 1:1~1.2; II. Preparation of mycelium powder: (1) Inoculate Bacillus saffron and Bacillus urealyticum onto tryptic soybean agar solid medium and incubate at 28-30℃ for 24-48 hours; (2) Take the strain cultured in step (1) and inoculate it into tryptic soybean liquid culture medium. Culture it at 28-30℃ for 24-48h to obtain seed liquid; (3) Inoculate the seed liquid into the fermentation medium at an inoculation rate of 2-3%, and ferment and culture at 28-30℃, 80-120rpm and pH 6.8-7.2 for 24-48h. Collect the fermentation liquid to obtain the microbial agent; spray the liquid into powder using a powder spraying device.

[0036] III. Preparation of oral liquid formulations: The herbal liquid is diluted 50% and sterilized. Then, 0.35% sodium chloride, 0.1% potassium chloride, 0.3% sodium bicarbonate, 1% glucose, and 0.5% glycine are added and mixed well. Finally, bacterial powder is added and mixed well to obtain the oral liquid preparation.

[0037] Comparative Example 1: This comparative example provides a fern extract. The fern is crushed to 50 mesh, placed in an extraction tank, soaked in 6 times the amount of 80% ethanol for 1 hour, then decocted at 60°C for 30 minutes, the supernatant is extracted, the ethanol is recovered by vacuum concentration, and the extract is boiled into an extract. Sterile ultrapure water is added to the extract, and impurities are filtered out to make it an aqueous solution with a concentration of 1:1~1.2.

[0038] Comparative Example 2: This comparative example provides a holly leaf extract, which is prepared by crushing holly leaves to 50 mesh, placing them in an extraction tank, adding 5 times the amount of water and soaking for 1 hour, then decocting at 75°C for 1.5 hours and extracting the supernatant; adding 5 times the amount of water a second time, decocting at 80°C for 2 hours, and extracting the supernatant; combining the supernatants and concentrating to 1 / 3 of the volume.

[0039] Comparative Example 3: This comparative example provides a traditional Chinese medicine liquid, which is a 1:1 mixture of extracts from *Pteris vittata* and *Ilex chinensis* leaves from Comparative Example 1.

[0040] Comparative Example 4: This comparative example provides a traditional Chinese medicine fermentation liquid, specifically by crushing and mixing fern, holly leaves and licorice root separately, adjusting the water content to about 60%, fermenting at 37°C for one week to complete the fermentation of traditional Chinese medicine, adding a compound bacterial solution of Bacillus spp. and Bacillus urealyticum and allowing it to stand and soak.

[0041] Comparative Example 5: This comparative example provides a traditional Chinese medicine oral liquid preparation, specifically by diluting the traditional Chinese medicine liquid prepared in Example 1 at a ratio of 50% and adding Bacillus urealyticum powder. The preparation process is the same as in Example 1, and a traditional Chinese medicine oral liquid preparation is made.

[0042] Comparative Example 6: This comparative example provides a traditional Chinese medicine oral liquid preparation, specifically by diluting the traditional Chinese medicine liquid prepared in Example 1 at a ratio of 50% and adding Bacillus sabinatus, and the preparation process is the same as in Example 1, to prepare a traditional Chinese medicine oral liquid preparation.

[0043] Experimental Example: To verify the effect of the oral liquid of traditional Chinese medicine prepared in this invention on pathogens, this experimental example uses the oral liquid preparation and traditional Chinese medicine liquid prepared in Example 1, as well as the medicinal liquid prepared in Comparative Examples 1-6, to conduct antibacterial tests on Escherichia coli, Salmonella, and Staphylococcus aureus pathogens, respectively. Escherichia coli was isolated from the feces of diarrheal piglets by the Hami City Animal Disease Prevention and Control Center; Salmonella was Salmonella typhimurium isolated from the feces of diarrheal calves in our laboratory; viable bacteria were counted using the turbidimetric method, and the resulting solution was prepared to a concentration of 1×10⁻⁶. 9 Pathogen suspension at cfu / mL; Staphylococcus aureus is preserved in our laboratory.

[0044] Use a pipette to aspirate 50 μL of 1 × 10⁻⁶ solution. 9The pathogen suspension at CFU / mL was evenly spread on the surface of agar medium. 6mm diameter circular blank antimicrobial susceptibility test discs were immersed in the solution for 1 hour and then air-dried on a clean bench for later use. The diameter of the inhibition zone for *Escherichia coli*, *Staphylococcus aureus*, and *Salmonella typhimurium* was determined using the disc method, with three replicates per group (judgment criteria: inhibition zone diameter <6mm indicates no antimicrobial effect; 6-9mm indicates mild antimicrobial effect; 10-20mm indicates moderate antimicrobial effect; and above 20mm indicates strong antimicrobial effect). The inhibitory effects of different groups of solutions on the pathogens are shown in Table 2 below.

[0045]

[0046] As shown in Table 2, both *Pteris vittata* and *Ilex chinensis* leaves exhibited certain inhibitory effects against *Escherichia coli*, *Salmonella*, and *Staphylococcus aureus*. However, compared to the prepared herbal liquid, the antibacterial effect of the single-component extracts was relatively weak, indicating that the combination of *Pteris vittata*, *Ilex chinensis* leaves, and licorice enhanced the antibacterial effect. The addition of bacterial powder significantly improved the antibacterial effect, suggesting that *Bacillus urealyticum* and *Bacillus safranin* promoted the antibacterial effect of the herbal liquid. Furthermore, the significantly better antibacterial effect of the oral liquid preparation prepared in this invention against *Escherichia coli* and *Salmonella* suggests that the synergistic effect of the two bacteria could enhance the antibacterial effect of the herbal liquid. This is likely because the bacterial powder can tolerate gastric acid and bile environments, allowing for rapid absorption in the intestines. The colonization process inhibits the proliferation of harmful bacteria such as Escherichia coli and Salmonella, stimulates intestinal mucosal immune cells, and enhances intestinal mucosal antibody levels and phagocytic capacity of immune cells. Licorice can reduce intestinal stress reactions that may be triggered in the early stages of colonization. The immunomodulatory effects of the bacterial powder and traditional Chinese medicine components are synergistic, improving the body's intestinal immunity. At the same time, the bacterial powder can also secrete active enzymes to improve the digestive and absorptive capacity of livestock and poultry while producing antibacterial peptides and other antibacterial components. It also forms a dual synergistic effect of biological and chemical antibacterial action with the antibacterial components of traditional Chinese medicine, enhancing the antibacterial effect and thoroughly addressing the two core causes of pathogenic bacterial infection and dysbiosis, reducing the risk of drug resistance that may arise from single-drug use.

[0047] Clinical application effects Case 1: On March 8, 2022, 15 one-month-old calves in a cattle farm with 300 head of cattle in Neihuang County developed diarrhea. The affected cattle had reduced appetite, were emaciated, had sunken eyes, watery feces, pale conjunctiva, thin white tongue coating, and deep pulse. They were diagnosed with neonatal diarrhea syndrome.

[0048] The preparation of this invention was administered orally at a dose of 50 ml per head, twice consecutively. The frequency of diarrhea was significantly reduced, and after two more doses, all cattle were completely cured. Consolidation medication was then given for 3 days, once a day. A follow-up visit 2 months later showed no recurrence, and the affected cattle showed a greater weight gain than other calves in the same batch.

[0049] Case 2: On February 20, 2023, some 45-day-old calves in a dairy farm with 1,000 cows in Jiyuan developed diarrhea. The affected calves were thin and had brittle coats, unsteady gait, loose and coarse feces mixed with some undigested food, large areas of loose feces adhered around the anus and the back of the thighs, pale mouth, soft tongue, weak pulse, and normal respiration and body temperature; the diagnosis was neonatal diarrhea syndrome.

[0050] The cow was given 60ml of the preparation of this invention orally three times. The diarrhea stopped and the cow's condition improved. The cow was given the medication twice more and the feces became formed. A follow-up visit two days later showed that the cow had fully recovered. A follow-up visit 30 days later showed that there was no recurrence.

[0051] Case 3: In a 500-pig farm in Zhumadian, some 5kg piglets fell ill. Affected pigs presented with white, watery feces with a sour odor, dirty anuses, chills, tendency to huddle together, lethargy, dull coat, reduced appetite, and some vomiting and cyanosis. The diagnosis was neonatal diarrhea syndrome. The treatment involved administering 10ml / pig of the preparation of this invention orally three times. The pigs' feces became thicker, and all symptoms improved. After two more treatments, they recovered completely. A follow-up visit two months later showed no recurrence of diarrhea symptoms.

[0052] Case 4: In a pig farm of 1000 pigs in Kaifeng, some piglets weighing 10kg fell ill. Affected pigs exhibited reduced or complete loss of appetite, some vomiting, followed by frequent diarrhea with thin or sticky, grayish, foul-smelling feces. They also showed lethargy, mild fever, scanty dark urine, and halitosis. The diagnosis was neonatal diarrhea syndrome. The pigs were treated with the preparation of this invention, 10ml per pig, administered orally for three consecutive times, resulting in complete recovery.

[0053] Case 5: In May 2022, at a sheep farm with 2000 sheep in Lankao County, some lambs developed diarrhea. The affected sheep had severe diarrhea, with watery, foul-smelling feces. The anus and tail base were heavily contaminated with feces containing sticky substances; the diagnosis was neonatal diarrhea syndrome. The sheep were treated with the preparation of this invention, 15ml per sheep, administered orally for three consecutive times, and recovered completely. A follow-up visit 15 days later showed that their appetite remained normal.

[0054] Case 6: On November 18, 2021, at a sheep farm in Zhoukou with 1,000 sheep, some 20-day-old lambs developed diarrhea. Affected lambs were lethargic, had sunken eyes, high skin temperature, and their hindquarters were contaminated with loose, brown, foul-smelling feces. They had no appetite but drank excessively. The diagnosis was neonatal diarrhea syndrome. The sheep were treated with the preparation of this invention, 10ml per lamb, administered orally three times. The diarrhea stopped, and their condition improved. One more dose was given, and their feces became formed. A follow-up visit two days later showed that the lambs had fully recovered.

[0055] The above description is only a preferred embodiment of the present invention and does not limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome, characterized in that: The raw materials include the following parts by weight: 3-6 parts of bracken fern, 3-6 parts of holly leaves, 5-10 parts of licorice, 1-3 parts of sodium chloride, 1-2 parts of potassium chloride, 1-3 parts of sodium bicarbonate, 1-8 parts of glucose, and 1-6 parts of glycine.

2. The traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome according to claim 1, characterized in that: The ingredients include the following parts by weight: 6 parts of bracken fern, 6 parts of holly leaves, 10 parts of licorice, 3 parts of sodium chloride, 2 parts of potassium chloride, 3 parts of sodium bicarbonate, 8 parts of glucose, and 6 parts of glycine.

3. The traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome according to claim 2, characterized in that: It also includes Bacillus salsa and Bacillus urealyticum powder.

4. The method for preparing a traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome according to any one of claims 1-3, characterized in that: Includes the following steps: Step 1: Preparation of the Traditional Chinese Medicine Solution, including the following steps: (1) Crush the fern into 50 mesh, put it into an extraction tank, add 6 times the amount of 80% ethanol and soak for 1 hour, then boil at 60°C for 30 minutes, extract the supernatant, concentrate under reduced pressure to recover the ethanol, boil into an extract, add sterile ultrapure water to the extract, filter out impurities, and obtain an aqueous solution with a concentration of 1:1~1.

2. (2) Crush the holly leaves to 50 mesh, put them into an extraction tank, add 5 times the amount of water and soak for 1 hour, then decoct at 75°C for 1.5 hours and extract the supernatant; add 5 times the amount of water a second time, decoct at 80°C for 2 hours and extract the supernatant; combine the supernatants and concentrate to 1 / 3 volume. (3) Add licorice to the extraction tank, add 5 times the amount of water and soak for 1 hour, then decoct at 100°C for 30 minutes and extract the supernatant; add 5 times the amount of water again, decoct at 80°C for 2 hours and extract the supernatant; combine the supernatants and concentrate to 1 / 3 volume. (4) Combine the three liquids, add sterile ultrapure water, filter to remove impurities, and make the concentration of the traditional Chinese medicine solution 1:1~1.2; Step 2, preparation of mycelium powder, includes the following steps: (1) Inoculate Bacillus saffron and Bacillus urealyticum onto tryptic soybean agar solid medium and incubate at 28-30 °C for 24-48 hours; (2) Take the strain cultured in step (1) and inoculate it into tryptic soybean liquid culture medium. Culture it at 28-30 ℃ for 24-48 h to obtain seed liquid; (3) Inoculate the seed liquid into the fermentation medium at an inoculation rate of 2-3%, and ferment and culture at 28-30 ℃, 80-120 rpm and pH 6.8-7.2 for 24-48 hours. Collect the fermentation liquid to obtain the microbial agent; spray the liquid into powder using a powder spraying device to obtain the product. Step 3: Preparation of oral liquid preparation: After diluting the Chinese medicine liquid according to the ratio, sterilize it, add 0.35% sodium chloride, 0.1% potassium chloride, 0.3% sodium bicarbonate, 1% glucose, and 0.5% glycine, and mix well; then add the bacterial powder and mix well to obtain the oral liquid preparation.

5. The method for preparing the traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome according to claim 3, characterized in that, In step three, the dilution ratio of the medicinal solution is 25%-50%.

6. The method for preparing the traditional Chinese medicine oral liquid preparation for treating neonatal diarrhea syndrome according to claim 3, characterized in that, The amount of Bacillus salsa in the oral liquid preparation is not less than 10. 6 cfu / mL.

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