Fermented traditional Chinese medicine composition for preventing and treating viral diarrhea of piglets as well as preparation method and application of fermented traditional Chinese medicine composition

Abstract

The invention relates to a fermented traditional Chinese medicine composition for preventing and treating piglet viral diarrhea as well as a preparation method and application thereof, and relates to the technical field of livestock breeding. The feed is prepared by taking traditional Chinese medicine components as basic raw materials and adding probiotics for fermentation, and the traditional Chinese medicine components comprise the following raw materials in parts by weight: 0.5-1.5 parts of cardamine violifolia dry powder, 5-15 parts of Chinese pulsatilla root, 10-20 parts of coptis chinensis, 10-20 parts of golden cypress, 10-20 parts of ash bark, 4-8 parts of turmeric, 6-10 parts of pumpkin seeds and 4-8 parts of a ginkgo leaf extract feed additive. Through cooperation of multiple components, the immunity and antioxidant functions of the body are enhanced, the structural integrity of the intestinal tract is maintained, piglet diarrhea caused by PEDV infection is effectively relieved, and growth and development of the piglet are promoted; local specialty crops are selected as medicines, and theoretical basis and practical reference are provided for deep processing of local genuine medicinal materials and PED prevention and control.

Description

Technical Field

[0001] This invention relates to the field of animal husbandry technology, and in particular to a fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets, its preparation method, and its application. Background Technology

[0002] Porcine epidemic diarrhea (PED) is an acute, highly contagious, and fatal intestinal infectious disease caused by porcine epidemic diarrhea virus (PEDV). Suckling piglets, whose immune and digestive systems are not yet fully developed, are the primary group affected. The younger the piglet, the higher the morbidity and mortality rates. PEDV belongs to the genus *A* coronaviruses in the family Coronaviridae. Its genome is a single-stranded positive-sense RNA, primarily infecting the epithelial cells of the pig small intestine. Viral replication within cells causes intestinal vacuolation and intestinal epithelial atrophy, disrupting the intestinal barrier function and thus interfering with the digestion and absorption of nutrients.

[0003] Currently, there is no specific drug treatment for PEDV infection in piglets. Furthermore, the virus continues to mutate, and existing prevention and control measures are gradually weakening in their protective effect on susceptible animals. Traditional Chinese veterinary medicine attributes PED to external pathogens invading the spleen and stomach, leading to internal dampness and toxins. Piglets are considered to have weak spleen and stomach functions, and improper feeding management or imbalances in temperature can easily cause spleen and stomach dysfunction, resulting in diarrhea. Therefore, treatment often employs principles such as clearing heat and detoxifying, strengthening the spleen and removing dampness, and astringing and stopping diarrhea. Multiple studies have confirmed that various traditional Chinese medicines, such as Coptis chinensis and Pulsatilla chinensis, have significant inhibitory effects on PEDV. Tannic acid and shikimic acid can improve the growth performance, intestinal barrier function, and antioxidant capacity of PEDV-infected newborn piglets. Andrographis paniculata decoction can resist PEDV infection by regulating the expression of IRF3 in the body and is less likely to induce drug resistance.

[0004] Currently, there are numerous research reports on the use of traditional Chinese medicine (TCM) to prevent and treat diarrhea in piglets. However, the raw materials for TCM are mostly plants, with high fiber content, low bioavailability, weak pharmacological effects, and a strong woody texture, which affects feed palatability and thus piglet feed intake. The therapeutic effect needs further improvement. To enhance the effectiveness and palatability of TCM preparations, the development of TCM microecological preparations has become a major research direction and hot topic. Probiotic fermented TCM utilizes modern fermentation engineering technology to inoculate beneficial microorganisms into a TCM matrix for growth and reproduction. Fermentation not only improves the palatability of the TCM but also increases the content and bioavailability of active ingredients, enhances efficacy, and reduces toxic side effects. Therefore, the development of fermented TCM compound feed additives has attracted much attention. Thus, the research and development of fermented TCM preparations specifically targeting the prevention and treatment of viral diarrhea in piglets has significant application prospects. Summary of the Invention

[0005] The purpose of this invention is to address the above-mentioned problems by providing a fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets, its preparation method, and its application.

[0006] The first aspect of this invention provides a method for preparing a fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets, employing the following technical solution:

[0007] A fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets is made by fermenting traditional Chinese medicine components with the addition of probiotics. The traditional Chinese medicine components include the following raw materials in parts by weight: 0.5-1.5 parts of dried powder of Viola yedoensis and Capsella bursa-pastoris, 5-15 parts of Pulsatilla chinensis, 10-20 parts of Coptis chinensis, 10-20 parts of Phellodendron chinense, 10-20 parts of Fraxinus chinensis, 4-8 parts of Curcuma longa, 6-10 parts of pumpkin seeds, and 4-8 parts of Ginkgo biloba extract feed additive.

[0008] By adopting the above technical solution, Viola yedoensis powder, turmeric, pumpkin seeds, and ginkgo leaves are added to the common Coptis chinensis and Pulsatilla chinensis formula. Viola yedoensis powder is prepared from fresh Viola yedoensis leaves and is rich in vitamin C, minerals, and glucosinolates. Fresh Viola yedoensis leaves from Enshi have a high selenium content. Sufficient selenium can significantly enhance the antioxidant capacity of piglets, effectively neutralize free radicals, reduce oxidative damage to intestinal mucosal cells caused by viruses, protect the integrity of the intestinal wall, and help improve the piglets' resistance to viruses, enabling the immune system to more effectively identify and eliminate invading viruses. When piglets have already developed viral diarrhea, the consumption of selenium in their bodies will increase dramatically. (Used to combat oxidative stress and inflammation), it may rapidly lead to selenium deficiency or a critical deficiency state, further weakening resistance and creating a vicious cycle. Timely and appropriate selenium supplementation through feeding during treatment can effectively combat severe oxidative stress caused by viruses, reduce intestinal cell damage, support immune system function, help the body clear viruses more quickly, promote the repair of damaged intestines, and reduce the risk of secondary infections (such as bacterial enteritis), thereby shortening the course of the disease, reducing mortality, and improving recovery. The core active ingredient in turmeric is curcumin, which has potent anti-inflammatory and antioxidant effects. Pumpkin seeds are rich in healthy fats, proteins, dietary fiber, and minerals. This product contains substances, vitamins, and antioxidants, possessing antioxidant and anti-inflammatory effects. Its rich vitamin E content is a fat-soluble antioxidant that blocks the chain reaction of lipid peroxidation in cell membranes. It has a synergistic effect with selenium, reducing intestinal oxidative damage and maintaining the integrity of the intestinal mucosal barrier. Selenium promotes lymphocyte proliferation, antibody (IgG / IgA) production, and enhances NK cell activity. Vitamin E improves T cell response, regulates Th1 / Th2 balance, and enhances mucosal immunity. Combined supplementation of both can improve piglets' ability to clear viruses, reduce secondary infections, and further reduce inflammatory responses (lowering TNF-α and IL-6) and accelerate... Ginkgo biloba extract promotes the regeneration of damaged intestinal epithelial cells, shortens the course of diarrhea, and is rich in flavonoids with antioxidant properties that protect cells from free radical damage. Its abundant terpene lactones have anti-platelet-activating factor effects, helping to improve blood circulation (especially microcirculation), reduce inflammation, and protect nerves. Feed additives containing curcumin, pumpkin seed, and ginkgo biloba extract exhibit strong neuroprotective potential through multiple mechanisms (anti-inflammatory, antioxidant, inhibiting amyloid aggregation, rich in minerals, neuronal protection, etc.) to avoid nerve damage caused by selenium supplementation. Combined with probiotic fermentation to improve palatability, this effectively alleviates diarrhea in piglets caused by PEDV infection while promoting growth and development.

[0009] Preferably, the traditional Chinese medicine components include the following raw materials in parts by weight: 1 part of dried Viola yedoensis leaf powder, 10 parts of Pulsatilla chinensis, 15 parts of Coptis chinensis, 15 parts of Phellodendron chinense, 15 parts of Fraxinus chinensis, 6 parts of Curcuma longa, 8 parts of pumpkin seeds, and 6 parts of Ginkgo biloba leaf extract feed additive.

[0010] Preferably, the selenium content of the dried Viola yedoensis powder is 900–1100 mg / kg, and more preferably 1000 mg / kg.

[0011] Preferably, the probiotic is a lyophilized lactic acid bacteria powder.

[0012] A second aspect of the present invention provides a method for preparing the above-mentioned fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets, using the following technical solution:

[0013] A method for preparing a fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets includes the following steps:

[0014] S1. Wash, dry, grind, and sieve the following ingredients: violet leaves, fresh water chestnut leaves, Pulsatilla chinensis, Coptis chinensis, Phellodendron chinense, Fraxinus chinensis, Curcuma longa, and pumpkin seeds.

[0015] S2. Weigh the raw materials obtained in S1 according to the ratio, add ginkgo leaf extract feed additive according to the ratio, stir and mix well, add probiotic freeze-dried powder, and ferment in a fermentation tank to obtain fermented Chinese medicine composition.

[0016] Preferably, in the above preparation method, the sieving operation in step S1 uses an 80-mesh sieve.

[0017] Preferably, in the above preparation method, the amount of probiotic freeze-dried powder added in S2 is 1% of the total mass of the traditional Chinese medicine components.

[0018] Preferably, in the above preparation method, the number of live bacteria in the probiotic freeze-dried powder in S2 is 1.0*10^6. 10 CFU / g.

[0019] Preferably, in the above preparation method, the fermentation temperature in step S2 is 36°C and the fermentation time is 6 days.

[0020] A third aspect of the present invention provides the use of the above-mentioned fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets in the preparation of a medicament for preventing and treating viral diarrhea in piglets.

[0021] In summary, this application includes at least one of the following beneficial technical effects:

[0022] 1. By adding Viola yedoensis powder, turmeric, pumpkin seeds and ginkgo leaves to the common Coptis chinensis and Pulsatilla chinensis formula, the multi-component synergistic effect enhances the body's immune and antioxidant functions, maintains the integrity of the intestinal structure, effectively relieves diarrhea in piglets caused by PEDV infection, and promotes their growth and development.

[0023] 2. By selecting local specialty crops for medicinal use, a theoretical basis and practical reference are provided for the deep processing of local medicinal materials and the control of PED.

[0024] 3. The preparation method is simple, the cost is low, and it is easy to promote. Detailed Implementation

[0025] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. Modifications or equivalent substitutions made by those skilled in the art based on their understanding of the technical solutions of this invention, without departing from the spirit and scope of the invention, should be covered within the protection scope of this invention.

[0026] The information on raw materials, reagents, instruments and equipment used in the following examples is as follows: all reagents and chemicals were used as is without further processing. Other specific conditions not specified were performed according to conventional conditions or the manufacturer's recommendations. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0027] Main materials:

[0028] The PEDV strain (number EZ202502) was isolated from a clinical sample from a pig farm in Enshi City, Hubei Province.

[0029] The selenium-enriched corydalis leaf and water chestnut were provided by the Enshi Academy of Agricultural Sciences.

[0030] Pulsatilla chinensis, Coptis chinensis, Phellodendron chinense, Fraxinus chinensis, Curcuma longa, and pumpkin seeds were all purchased from Enshi Yuanchang Pharmaceutical Chain Co., Ltd.

[0031] Ginkgo biloba extract feed additive was purchased from Hebei Yulongxiang Biotechnology Co., Ltd.

[0032] The lyophilized lactic acid bacteria powder was purchased from Henan Huachu Biotechnology Co., Ltd.

[0033] The premixed feed and puffed soybeans were purchased from Badong County Huifeng Animal Husbandry Co., Ltd.

[0034] Main instruments and equipment:

[0035] The embedding machine, freezing table, and tissue spreader were purchased from Wuhan Junjie Electronics Co., Ltd.

[0036] The pathology slide machine was purchased from Shanghai Leica Instruments Co., Ltd.

[0037] The optical microscope and imaging system were purchased from Nikon Corporation of Japan.

[0038] I. Implementation Examples

[0039] Example 1

[0040] A fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets is made by fermenting traditional Chinese medicine components with the addition of probiotics. The traditional Chinese medicine components include the following raw materials in parts by weight: 1 part of dried powder of violet leaves and crushed rice, 10 parts of Pulsatilla chinensis, 15 parts of Coptis chinensis, 15 parts of Phellodendron chinense, 15 parts of Fraxinus chinensis, 6 parts of Curcuma longa, 8 parts of pumpkin seeds, and 6 parts of Ginkgo biloba extract feed additive.

[0041] A method for preparing a fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets includes the following steps:

[0042] S1. Wash, dry, grind, and sieve the following ingredients: fresh leaves of violet, crushed rice, pulverized stalks, coptis, phellodendron, fraxinus bark, turmeric, and pumpkin seeds.

[0043] Specifically, 20 kg of fresh leaves of *Vigna purpurea* cultivated by the Enshi Academy of Agricultural Sciences were taken. The selenium content was between 900 and 1100 mg / kg, with an average selenium content of 1000 mg / kg. The fresh leaves were washed, dried, pulverized, and passed through an 80-mesh sieve to obtain dried *Vigna purpurea* powder for later use. *Pulsatilla chinensis*, *Coptis chinensis*, *Phellodendron chinense*, *Fraxinus chinensis*, turmeric, and pumpkin seeds were washed, dried, pulverized, and passed through an 80-mesh sieve for later use.

[0044] S2. Weigh the raw materials obtained in S1 according to the ratio, add ginkgo leaf extract feed additive according to the ratio, stir and mix well, add probiotic freeze-dried powder, and ferment in a fermentation tank to obtain fermented Chinese medicine composition.

[0045] Specifically, 100g of dried Viola yedoensis leaf powder, 1000g of Pulsatilla chinensis, 1500g of Coptis chinensis, 1500g of Phellodendron chinense, 1500g of Fraxinus chinensis, 600g of Curcuma longa, and 800g of pumpkin seeds were weighed out from S1. 600g of Ginkgo biloba extract feed additive was added and mixed thoroughly. Lactic acid bacteria freeze-dried powder (with a live bacteria count of 1.0 × 10¹) was added at a ratio of 100:1 based on the total mass of the Chinese herbal powder. 0 The CFU / g of the herbal medicine was placed in a solid-state fermentation tank and fermented at 36°C for 6 days to obtain a fermented yellow herbal medicine composition for later use.

[0046] In this embodiment, the probiotic freeze-dried powder is lactic acid bacteria freeze-dried powder. In other feasible embodiments, Lactobacillus casei or Lactobacillus acidophilus can also be selected as substitutes.

[0047] Example 2

[0048] A fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets differs from Example 1 in that the traditional Chinese medicine components include the following raw materials in parts by weight: 0.5 parts of dried Viola yedoensis leaf powder, 5 parts of Pulsatilla chinensis, 10 parts of Coptis chinensis, 10 parts of Phellodendron chinense, 10 parts of Fraxinus chinensis, 4 parts of Curcuma longa, 6 parts of pumpkin seeds, and 4 parts of Ginkgo biloba leaf extract feed additive.

[0049] The preparation method of a fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets is basically the same as that in Example 1, except that the dosage of each component is adjusted, which will not be repeated here.

[0050] Example 3

[0051] A fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets differs from Example 1 in that the traditional Chinese medicine components include the following raw materials in parts by weight: 1.5 parts of dried Viola yedoensis leaf powder, 15 parts of Pulsatilla chinensis, 20 parts of Coptis chinensis, 20 parts of Phellodendron chinense, 20 parts of Fraxinus chinensis, 8 parts of Curcuma longa, 10 parts of pumpkin seeds, and 8 parts of Ginkgo biloba leaf extract feed additive.

[0052] The preparation method of a fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets is basically the same as that in Example 1, except that the dosage of each component is adjusted, which will not be repeated here.

[0053] II. Comparative Example

[0054] Comparative Example 1

[0055] Preparation method of dried Viola yedoensis leaf powder: Take 20 kg of fresh Viola yedoensis leaf powder cultivated by Enshi Academy of Agricultural Sciences. The selenium content is 1000 mg / kg. After washing and drying, the fresh leaves are crushed and passed through an 80-mesh sieve to obtain dried Viola yedoensis leaf powder for later use.

[0056] Comparative Example 2

[0057] A fermented Coptis chinensis formula is prepared according to the following method: 1000 g of Pulsatilla chinensis, 1500 g of Coptis chinensis, 1500 g of Phellodendron chinense, and 1500 g of Fraxinus chinensis are weighed, pulverized, and passed through an 80-mesh sieve, then mixed evenly; lyophilized lactic acid bacteria powder (with a viable count of 1.0 × 10¹) is added at a ratio of 100:1 of the total mass of the Chinese herbal powder. 0 The CFU / g of the extract was placed in a solid-state fermentation tank and fermented at 36°C for 5 days to obtain the fermented Coptis chinensis formula for later use.

[0058] III. Performance Test Experiments and Results

[0059] 1. Experimental animals and grouping

[0060] Twenty-four paternal half-sib piglets (Duroc × Landrace × Large White three-way cross) aged 7 days, in good health, and with a weight of approximately (2.65±0.11kg) were selected. The animals were sourced from the Enshi branch of Yichang Zhengda Livestock Co., Ltd., and were randomly divided into 4 groups of 6 piglets each, with 3 replicates and 2 piglets per replicate.

[0061] The experimental animals were divided into four groups: the PEDV group served as the challenge control; the selenium-enriched group (Se-PEDV) was supplemented with selenium-enriched Corydalis yanhusuo prepared in Comparative Example 1 in the basal diet; the fermented Chinese medicine group (Coptis-PEDV) was supplemented with fermented Coptis chinensis prepared in Comparative Example 2 in the basal diet; and the selenium-enriched fermented Chinese medicine group (Se-Coptis-PEDV) was supplemented with the fermented Chinese medicine composition prepared in Example 1 in the basal diet.

[0062] All groups were fed artificial milk replacer under the same feeding and management conditions. The formula was prepared according to the method of Li Huanhuan et al. (see Reference 1 for details). The additives were added at the following amounts per 10 kg of artificial milk replacer and starter feed: 3.56 g of selenium-enriched violet leaf fragments prepared in Comparative Example 1 were added to the selenium-enriched group; 200 g of fermented coptis chinensis formula prepared in Comparative Example 2 were added to the fermented Chinese medicine formula prepared in Comparative Example 2; and 200 g of fermented Chinese medicine composition prepared in Example 1 were added to the selenium-enriched fermented Chinese medicine group. The composition of the basic starter feed during the experimental stage is shown in Table 1.

[0063] Table 1. Composition and nutrient levels of basic starter feed

[0064] Raw material composition / ‰ content Nutritional level content Expanded corn 315.25 Crude protein (%) 18.74 puffed rice 160 Crude fat (%) 5.54 puffed wheat germ 100 Crude fiber (%) 1.38 High-protein whey powder 62.5 Coarse ash content (%) 8.14 Low-protein whey powder 62.5 calcium(%) 0.59 puffed soybeans 40 Total phosphorus (%) 0.61 Imported fishmeal 40 Non-phytic acid phosphorus (%) 0.46 Fermented soybean meal 40 Salt % (%) 0.54 White sugar 30 Pig DE (kcal / kg) 3,625.20 Milk fat energy 30 Pig NE (kcal / kg) 2,702.60 porcine intestinal membrane protein powder 25 Lysine (%) 1.54 Soy protein hydrolysate 20 SID protein + cysteine ​​ / SID lysine (%) 0.6 Soy protein concentrate 15 - - Soybean oil 18 - - Calcium hydrogen phosphate 9 - - MS0511T(20kg) 5 - - VS0511T (20kg) 5 - - benzoic acid 4 - - L-Lysine hydrochloride 3 - - threonine 2 - - Zinc oxide 2 - - fumaric acid 2 - - fine stone powder 2 - - Myristic acid 1.8 - - DL-methionine 1.3 - - Sodium chloride 1.2 - - 50% choline chloride 0.7 - - Valine 0.5 - - Tryptophan 0.4 - - 9% Aminoferric 0.4 - - Fragrance 0.4 - - antioxidants 0.3 - - sweeteners 0.25 - - High-temperature phytase 0.2 - - essential oils 0.3 - - total 1000 - -

[0065] 2. Sample Collection and Processing

[0066] On day 15 of the experiment, three piglets were randomly selected from each group, and 20 mL of blood was collected from the anterior vena cava. After the blood samples were left to stand for 30 minutes, the serum was separated by centrifugation, aliquoted, and 3 mL was stored at -80℃ for later use. The piglets were anesthetized by intravenous injection of propofol, euthanized by potassium chloride, and then necropsy was performed.

[0067] A 2 cm segment of duodenal tissue was harvested. One portion was fixed with 4% paraformaldehyde solution for subsequent morphological analysis. The other portion was quickly placed in a cryovial, flash-frozen in liquid nitrogen, and then transferred to an ultra-low temperature freezer at -80°C for storage for molecular biological assays. At the same time, jejunal contents were collected, flash-frozen in liquid nitrogen, and also stored at -80°C for later use.

[0068] 3. Indicator Measurement and Result Analysis

[0069] 3.1 Diarrhea rate in piglets

[0070] During the initial 3-day acclimatization period of the experiment, the diarrhea rate of piglets in all groups was 0% by day 3. On day 4, piglets were orally administered PEDV, and their diarrhea, feed intake, mental state, and fecal color were recorded. The diarrhea rate of each group of piglets was calculated using the following formula:

[0071] Diarrhea rate (%) = (Number of piglets with diarrhea / Total number of piglets in each group) * 100%

[0072] The results are as follows: Starting from day 5, clinical symptoms such as diarrhea, loss of appetite, and weight loss appeared successively in all groups. On day 7, before the administration of the fermented Chinese medicine composition, the diarrhea rate in the PEDV group, Se-PEDV group, Coptis-PEDV group, and Se-Coptis-PEDV group was 100%, proving that the PEDV model was successfully established.

[0073] Starting from day 7, the Se-PEDV group, Coptis-PEDV group, and Se-Coptis-PEDV group were administered suspensions prepared with the corresponding additives and artificial milk via gavage. By day 15, before slaughter, the diarrhea rates of each group were as follows: PEDV group 100%, Se-PEDV group 83.33%, Coptis-PEDV group 33.33%, and Se-Coptis-PEDV group 16.67%.

[0074] This demonstrates that the fermented traditional Chinese medicine composition provided by this method can significantly reduce the diarrhea rate in PEDV-infected piglets.

[0075] 3.2 Daily weight gain of piglets

[0076] On the first day of the experiment, the weight of each group of piglets was recorded as the initial weight (kg). On the 14th day of the experiment, water and feed were stopped at 17:00. On the morning of the 15th day of the experiment, the fasting weight of each group of piglets was measured and recorded as the final weight (kg). The daily weight gain of each group of piglets was calculated using the following formula:

[0077] Daily weight gain = ((final weight - initial weight) / number of days of trial)

[0078] The daily weight gain results for each group are shown in Table 2:

[0079] Table 2 Comparison of Daily Weight Gain of Piglets

[0080] project PEDV Se-PEDV Coptis-PEDV Se-Coptis-PEDV Initial weight (kg) 2.55±0.02 2.54±0.03 2.53±0.02 2.55±0.01 Final weight (kg) 3.82±0.12 3.97±0.13 4.49±0.11 4.59±0.14 Daily weight gain (g) 84.89±3.39 95.11±6.98 129.00±7.03 137.33±8.80

[0081] As shown in Table 2, the daily weight gain of the Se-PEDV group was significantly higher than that of the PEDV group (P < 0.05); while the daily weight gain of the Coptis-PEDV group and the Se-Coptis-PEDV group was significantly higher than that of the PEDV group and the Se-PEDV group (P < 0.05), but the difference between the two groups was not significant (P > 0.05).

[0082] This demonstrates that the fermented traditional Chinese medicine composition provided by this method can effectively increase the daily weight gain of PEDV-infected piglets.

[0083] 3.3 Serum biochemical indicators

[0084] Serum samples were collected and used to determine blood immune indicators in different groups of piglets: immunoglobulin G (IgG), interleukin-2 (IL-2) and serum antioxidant indicators: superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), and malondialdehyde (MDA). Kits were purchased from Norminkoda, and the detection instrument was a full-band ELISA reader. The assay methods followed the instructions on the kits and standard detection methods.

[0085] The test results are shown in Table 3:

[0086] Table 3. Results of serum biochemical marker detection

[0087] project PEDV Se-PEDV Coptis-PEDV Se-Coptis-PEDV Immunoglobulin G (IgG) (μg / mL) 230.40±5.79 258.43±9.78 319.58±13.6 376.51±11.96 Interleukin-2 (IL-2) (pg / mL) 334.12±10.35 333.30±29.15 412.13±11.50 471.84±21.39 Superoxide dismutase (SOD) (U / mL) 24.48±0.42 26.07±0.40 39.13±0.52 40.08±3.22 Total antioxidant capacity (T-AOC) (μmol / mL) 0.95±0.12 1.26±0.13 1.82±0.13 1.83±0.15 Glutathione peroxidase (GPX) (U / mL) 258.88±22.40 264.05±18.58 353.64±9.28 365.43±22.19 Malondialdehyde (MDA) (nmol / mL) 1.22±0.11 1.39±0.24 1.06±0.08 0.93±0.01

[0088] Table 3 shows that, in terms of improving serum immune and antioxidant indicators, the fermented Chinese herbal medicine groups (Coptis-PEDV and Se-Coptis-PEDV) were more effective than the single selenium-enriched group (Se-PEDV). Specifically: compared with the PEDV group, the Se-PEDV group showed significant increases only in IgG and T-AOC; the Coptis-PEDV group showed significant increases in IgG, T-AOC, IL-2, SOD, and GPX; and the Se-Coptis-PEDV group showed significant improvements in all indicators. Compared with the Se-PEDV group, the Coptis-PEDV group showed more significant increases in IgG, T-AOC, IL-2, SOD, and GPX (P<0.05). Compared with the Coptis-PEDV group, the improvement effect of the Se-Coptis-PEDV group was mainly concentrated in the further increase of IgG and IL-2 (P<0.05), while no significant differences were found in T-AOC, SOD, GPX, and MDA.

[0089] This demonstrates that the fermented traditional Chinese medicine composition provided by this method can effectively increase the activities of T-AOC, SOD and GSH-Px in the serum of PEDV-infected piglets, reduce the MDA content, and thus alleviate oxidative stress and tissue damage caused by PEDV infection.

[0090] 3.4 Morphology and Structure of the Small Intestinal Mucosa

[0091] Duodenal samples were embedded in paraffin and stained with safranin and fast green. They were then observed using a Nikon ECLIPSE CI upright optical microscope and FI3 imaging system to examine the duodenal mucosal structure, measure the height of small intestinal villi and the depth of crypts, and calculate the average values ​​of each indicator.

[0092] The test results are shown in Table 4:

[0093] Table 4. Effects of different groups on duodenal injury in PEDV-infected piglets.

[0094] project PEDV Se-PEDV Coptis-PEDV Se-Coptis-PEDV Intestinal villus length (μm) 274.40±35.63 383.22±44.76 340.97±30.14 486.57±56.50 crypt depth (μm) 328.27±50.33 324.12±38.77 344.50±79.04 246.22±59.85

[0095] Table 4 shows that different treatments had different effects on villus length and crypt depth. The intergroup comparison of villus length showed that the Coptis-PEDV group and the Se-Coptis-PEDV group were different from the PEDV group and the Se-PEDV group (P<0.05), and the villus length of the Se-Coptis-PEDV group was significantly better than that of the Coptis-PEDV group (P<0.05). As for the crypt depth, the Se-Coptis-PEDV group was significantly lower than the other groups (P<0.05).

[0096] The small intestine is the main site for the digestion and absorption of nutrients in animals. When piglets are infected with PEDV, they often show pathological changes such as damage and loss of intestinal villi, resulting in a significant decrease in the ratio of villi height to crypt depth. The results of this experiment further confirm that the fermented traditional Chinese medicine composition provided by this method can significantly increase villi height, reduce crypt depth, and maintain the integrity of intestinal structure in PEDV-infected piglets.

[0097] The fermented traditional Chinese medicine compositions prepared in Examples 2 and 3 were tested repeatedly, and the results of each test were basically consistent with the results above, so they will not be repeated here.

[0098] In summary, this method, by adding dried powder of Viola yedoensis, Citrus aurantium, turmeric, pumpkin seeds, and Ginkgo biloba leaves to the common Coptis chinensis and Pulsatilla chinensis formula, effectively alleviates diarrhea in piglets caused by PEDV infection and promotes their growth and development through the synergistic effect of multiple components to enhance the body's immune and antioxidant functions and maintain the integrity of the intestinal structure. Furthermore, by using local specialty crops as medicine, it provides a theoretical basis and practical reference for the deep processing of local medicinal materials and the prevention and control of PEDV. The preparation method is simple, low-cost, and easy to promote, providing a new approach to the prevention and treatment of diarrhea in piglets caused by PEDV infection.

[0099] The references mentioned in this application are as follows:

[0100] [1] Li Huanhuan, Yu Chenmin, Tian Xiaorong, et al. Protective effect of catechu extract on colon infection of young piglets with porcine epidemic diarrhea virus [J]. Chinese Journal of Animal Husbandry and Veterinary Medicine, 2025, 52(03):1360-1369. DOI:10.16431 / j.cnki.1671-7236.2025.03.037.

Claims

1. A fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets, characterized in that: It is made from traditional Chinese medicine components as the base raw materials and fermented with probiotics. The traditional Chinese medicine components include the following raw materials in parts by weight: 0.5-1.5 parts of dried Viola yedoensis leaf powder, 5-15 parts of Pulsatilla chinensis, 10-20 parts of Coptis chinensis, 10-20 parts of Phellodendron chinense, 10-20 parts of Fraxinus chinensis, 4-8 parts of Curcuma longa, 6-10 parts of pumpkin seeds, and 4-8 parts of Ginkgo biloba leaf extract feed additive.

2. The fermented traditional Chinese medicine composition according to claim 1, characterized in that: The traditional Chinese medicine components include the following raw materials in parts by weight: 1 part of dried Viola yedoensis leaf powder, 10 parts of Pulsatilla chinensis, 15 parts of Coptis chinensis, 15 parts of Phellodendron chinense, 15 parts of Fraxinus chinensis, 6 parts of Curcuma longa, 8 parts of pumpkin seeds, and 6 parts of Ginkgo biloba leaf extract feed additive.

3. The fermented traditional Chinese medicine composition according to claim 1, characterized in that: The selenium content of the dried Viola yedoensis and Citrus aurantium powder is 900–1100 mg / kg, preferably 1000 mg / kg.

4. The fermented traditional Chinese medicine composition according to claim 1, characterized in that: The probiotics are freeze-dried lactic acid bacteria powder.

5. The method for preparing the fermented traditional Chinese medicine composition for preventing and treating viral diarrhea in piglets according to any one of claims 1-4, characterized in that: Includes the following steps: S1. Wash, dry, grind, and sieve the following ingredients: violet leaves, fresh water chestnut leaves, Pulsatilla chinensis, Coptis chinensis, Phellodendron chinense, Fraxinus chinensis, Curcuma longa, and pumpkin seeds. S2. Weigh the raw materials obtained in S1 according to the ratio, add ginkgo leaf extract feed additive according to the ratio, stir and mix well, add probiotic freeze-dried powder, and ferment in a fermentation tank to obtain fermented Chinese medicine composition.

6. The preparation method according to claim 5, characterized in that: In S1, an 80-mesh sieve is selected for the sieving operation.

7. The preparation method according to claim 5, characterized in that: The amount of probiotic freeze-dried powder added in S2 is 1% of the total mass of the traditional Chinese medicine components.

8. The preparation method according to claim 7, characterized in that: The live bacteria count in the probiotic freeze-dried powder in S2 is 1.0*10^6 bacteria. 10 CFU / g.

9. The preparation method according to claim 7, characterized in that: The fermentation temperature in S2 is 36℃, and the fermentation time is 6 days.

10. The use of the fermented traditional Chinese medicine composition according to any one of claims 1-4 in the preparation of a drug for preventing and treating viral diarrhea in piglets.