Application of hesperetin in preparing medicament or feed additive for treating or preventing porcine cystovirus infection

Abstract

The present application discloses the use of hesperetin in the preparation of a drug or feed additive for treating or preventing porcine enveloped virus infection. The drug is hesperetin or an oral liquid, granule, tablet, or powder prepared by adding a solvent or excipient with hesperetin as the active ingredient, and the dosage is 2-10 mg / kg of pig body weight; or hesperetin is directly added to the feed, and the addition amount is 0.01%-0.03% of the total weight of the pig feed. The present application uses hesperetin or a composition including hesperetin as a drug or feed additive, which has high safety, no toxic and side effects, and has a significant effect of inhibiting the replication of enveloped viruses such as porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, and African swine fever virus, and reducing the virulence of porcine enveloped virus. It overcomes the disadvantages such as immune failure that may occur after vaccination and virus mutation caused by the use of antiviral drugs, and effectively reduces the incidence and mortality of piglets infected with porcine enveloped virus.

Description

Technical Field

[0001] This application relates to the technical field of prevention and control of porcine enveloped viruses, and specifically relates to the application of hesperetin in the preparation of drugs or feed additives for treating or preventing porcine enveloped virus infections. Background Art

[0002] Porcine enveloped viruses refer to a class of porcine viruses with an envelope structure, which can cause various diseases in pigs. Currently, common porcine enveloped viruses include porcine epidemic diarrhea virus (PEDV), porcine reproductive and respiratory syndrome virus (PRRSV), African swine fever virus (ASFV), etc. PEDV is the main cause of porcine epidemic diarrhea (PED). After entering the body through the digestive tract, PEDV can enter cells for replication and proliferation by binding to receptors, and antagonize the host's innate immune response, causing immune suppression in the body. PEDV mainly causes vomiting and watery diarrhea in neonatal piglets within 10 days of age, leading to high morbidity and mortality rates in neonatal piglets. PRRSV infection can cause porcine reproductive and respiratory syndrome (PRRS). This virus mainly infects porcine alveolar macrophages, resulting in disorders of the immune system function, mainly manifested as low levels and short durations of neutralizing antibody responses; suppression of cellular immune function, low and delayed expression levels of interferon-γ; and infected pigs continuously carrying the virus, making it difficult to eliminate the virus from the pig herd. ASFV is the pathogen that causes African swine fever (ASF). ASF is a highly pathogenic, highly lethal, and highly contagious animal disease, with a mortality rate of up to 100% in its most acute form, and it is one of the major animal infectious diseases that must be reported as required by the World Organization for Animal Health (OIE).

[0003] PEDV, PRRSV, and ASFV have caused the death of a large number of piglets, resulting in huge economic losses and seriously hindering the development of the pig breeding industry. Currently, the prevention and control measures for PEDV, PRRSV, and ASFV virus infection diseases in China mainly rely on vaccines. However, due to the abuse of large doses of antiviral and antibacterial drugs, the mutation speed of virus (bacteria) strains far exceeds the speed of vaccine development, and the immune suppression characteristics of the virus lead to poor vaccine immune effects. In addition, since vaccine immunization only has the effect of preventing the occurrence of viral diseases and cannot provide protection for piglets that have been infected and developed the disease. Therefore, relying solely on vaccines easily leads to immune failure, and large-scale PEDV, PRRSV, and ASFV virus infection diseases still break out frequently.

[0004] In view of the problems of existing vaccines in preventing and controlling porcine cystovirus, the development of functional nutrients with anti-porcine cystovirus will have broad application prospects. Summary of the Invention

[0005] In view of this, the purpose of this application is to provide the application of hesperetin in the preparation of drugs or feed additives for treating or preventing porcine cystovirus infection. This hesperetin is a natural additive, which has the effects of anti-porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus and African swine fever virus, with high safety and convenient application. This hesperetin can be used to prepare drugs or feed additives for treating or preventing porcine cystovirus infection.

[0006] To achieve or at least partially achieve the above purpose, this application provides the following technical solutions:

[0007] The application of hesperetin in the preparation of drugs or feed additives for treating or preventing porcine cystovirus infection.

[0008] The technical solutions provided by this application, compared with the existing technologies, have at least the following beneficial effects:

[0009] 1. The hesperetin or the preparation containing hesperetin provided by this application can be used as a feed additive for a long time to achieve the purpose of preventing diseases. Long-term use will not cause the generation of drug-resistant strains, nor will there be other toxic and side effects, overcoming the disadvantages such as immune failure that may occur during vaccination and virus mutation caused by the use of antiviral drugs; after feeding this hesperetin or the preparation containing hesperetin to pigs, it can enhance the body's antioxidant defense function, improve the natural immune function and antiviral activity of pigs, and effectively prevent and control the infection of porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus and African swine fever virus.

[0010] 2. The in vitro experimental results of this application prove that hesperetin has a significant effect of inhibiting the proliferation of viruses such as PEDV, PRRSV and ASFV.

[0011] 3. The in vivo experimental results of this application show that hesperetin has the functions of enhancing natural immune function, antiviral activity, improving the intestinal structure and absorption function of pigs, can effectively prevent and control the infection of porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, African swine fever virus, and effectively improve the survival rate and production performance of pigs. The protection rate against the infection of porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, African swine fever virus can reach more than 90% by orally administering hesperetin at 10 mg / kg of pig body weight or adding hesperetin to the feed of piglets. Detailed Embodiments

[0012] To make the objectives, technical solutions and advantages of the present application more clear and understandable, the present application will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0013] Those skilled in the art of this technology can understand that unless specifically stated, the words "the", "this", "the foregoing" used in the text of the present application may also include plural forms. It should be further understood that the term "including" used in the specification of the present application means the presence of the described features, steps, operations, but does not exclude the presence or addition of one or more other features, integers, steps.

[0014] Those skilled in the art of this technology can understand that for those embodiments where specific experimental steps or conditions are not indicated, the operations or conditions of the conventional experimental steps described in the literature in this field can be followed; for those raw materials or instrument and equipment where the manufacturers are not indicated, they are all conventional products that can be obtained through commercial purchase.

[0015] Those skilled in the art of this technology can understand that unless otherwise stated in the present application, when an embodiment gives a numerical range, any value between the two endpoints of each numerical range and any value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present application, based on the understanding of those skilled in the art of this technology of the prior art and the description of the present application, any method, equipment and material similar or equivalent to the methods, equipment and materials described in the embodiments of the present application can also be used to implement the present application.

[0016] Hesperetin (HES) is a well-known and common flavonoid glycoside traditional Chinese medicine (TCM), and its molecular formula is C 16 H 14 O6. Its appearance is usually light yellow needle-shaped crystals or powder, and some also say it is a beige to light brown crystalline solid. HES has a relatively high content in medicinal materials such as Fructus Aurantii Immaturus, Fructus Aurantii, and Pericarpium Citri Reticulatae, and mostly exists in Rutaceae plants in the form of glycosides.

[0017] Through in vivo and in vitro experiments, the embodiments of the present application found that hesperetin and the composition containing hesperetin can significantly inhibit the proliferation of PEDV, PRRSV and ASFV, as well as inhibit their infection of cells. This hesperetin and the composition containing hesperetin (hereinafter referred to as "hesperetin composition") can be used to prepare drugs or feed additives against porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, and African swine fever virus.

[0018] Based on this, the embodiments of the present application provide the application of hesperetin in the preparation of drugs or feed additives for the treatment or prevention of porcine enveloped virus infection.

[0019] In some embodiments, the porcine virus includes at least one of porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, and African swine fever virus.

[0020] In some embodiments, the drug or feed additive is hesperetin, which can be directly used as a drug or feed additive for treating or preventing porcine diseases such as PED, PRRS, and ASF.

[0021] In some embodiments, the drug or feed additive is a hesperetin composition, and the hesperetin composition includes hesperetin and excipients, and the excipients include at least one of absolute ethanol, water, Tween-80, lactose, defatted rice bran, corn cob powder, soluble starch, wheat bran, silica, montmorillonite, vermiculite, medical stone, zeolite powder, and white carbon black.

[0022] In some embodiments, the aforementioned hesperetin composition is a solution formed by mixing hesperetin with three solvents, namely absolute ethanol, Tween-80, and water, wherein the concentration of hesperetin is 20 - 100 mg / mL.

[0023] In some embodiments, the aforementioned hesperetin composition is a solid mixture formed by mixing hesperetin with lactose and soluble starch, wherein the mass fraction of hesperetin in the solid mixture is 45% - 55%.

[0024] In some embodiments, the aforementioned hesperetin composition is a powdery mixture formed by mixing hesperetin with soluble starch or wheat bran, wherein the mass fraction of hesperetin in the powdery mixture is 45% - 55%.

[0025] In some embodiments, when the aforementioned drug or feed additive is a hesperetin composition, its preparation types include at least one of oral liquid, granule, tablet, and powder, and the preparation of the preparation can be carried out by conventional pharmaceutical preparation methods.

[0026] In some embodiments, the usage methods of the drug or feed additive include at least one of the following methods:

[0027] (1) Adding hesperetin or hesperetin composition to feed or drinking water for use according to a dose of 2 - 10 mg / kg of pig body weight in terms of hesperetin content;

[0028] (2) Directly orally administering hesperetin or hesperetin composition according to a dose of 2 - 10 mg / kg of pig body weight in terms of hesperetin content;

[0029] (3) Adding hesperetin or hesperetin composition to feed for use according to a dose of 0.01% - 0.03% of the total weight of pig feed in terms of hesperetin content.

[0030] The technical solutions and achieved technical effects of the present application will be described in detail through more specific embodiments below.

[0031] Example 1

[0032] This example provides a preparation method of hesperetin oral liquid against porcine capsid virus. The specific steps include:

[0033] (1) Weigh 2 g of hesperetin, 20 mL of absolute ethanol, 30 mL of Tween-80, and an appropriate amount of injection water;

[0034] (2) First dissolve hesperetin in absolute ethanol, then add Tween-80, and finally slowly add injection water to make up to 100 mL; mix evenly to obtain a 20 mg / mL hesperetin solution, that is, the hesperetin oral liquid C1.

[0035] Example 2

[0036] This example provides a preparation method of hesperetin oral liquid against porcine capsid virus. The specific steps include:

[0037] (1) Weigh 10 g of hesperetin, 20 mL of absolute ethanol, 30 mL of Tween-80, and an appropriate amount of injection water;

[0038] (2) First dissolve hesperetin in absolute ethanol, then add Tween-80, and finally slowly add injection water to make up to 100 mL; mix evenly to obtain a 100 mg / mL hesperetin solution, that is, the hesperetin oral liquid C2.

[0039] Example 3

[0040] This example provides a preparation method of hesperetin granule against porcine capsid virus. The specific steps include:

[0041] (1) Weigh 50 g of hesperetin, 25 g of lactose, and 25 g of soluble starch for later use;

[0042] (2) Mix hesperetin, lactose, and soluble starch to granulate and dry to obtain a granule mixture with a hesperetin mass fraction of 50%, that is, the hesperetin granule C3.

[0043] Example 4

[0044] This example provides a preparation method of hesperetin powder against porcine capsid virus. The specific steps include:

[0045] (1) Weigh 50 g of hesperetin and 50 g of wheat bran for later use;

[0046] (2) Mix hesperetin and wheat bran evenly and pass through a 60-mesh sieve to obtain a powdery mixture with a hesperetin mass fraction of 50%, that is, the hesperetin powder C4.

[0047] Example 5

[0048] This embodiment provides a method for preparing hesperetin tablets against porcine enveloped virus, and the specific steps include:

[0049] (1) Weigh 50 g of hesperetin, 25 g of lactose, and 25 g of soluble starch for later use.

[0050] (2) After uniformly mixing hesperetin, lactose, and soluble starch, press and dry them with a tableting machine to obtain a flaky mixture with a mass fraction of hesperetin of 50%, that is, the hesperetin tablet C5.

[0051] The formula and parameters of the hesperetin preparation prepared in the above embodiment are shown in Table 1 below:

[0052] Table 1

[0053] Example Number Preparation Number Preparation Type Components Concentration of Chebulinic Acid Chelate Example 1 C1 Oral Liquid Hesperetin, Absolute Ethanol, Tween-80 20mg / mL Example 2 C2 Oral Liquid Hesperetin, Absolute Ethanol, Tween-80 100mg / mL Example 3 C3 Granules Hesperetin, Lactose, Soluble Starch 50%(mass fraction) Example 4 C4 Powder Hesperetin, Bran 50%(mass fraction) Example 5 C5 Tablets Hesperetin, Lactose, Soluble Starch 50%(mass fraction)

[0054] Inhibitory Effect of Hesperetin on the In vitro Proliferation of PEDV, PRRSV and ASFV

[0055] In the embodiment of this application, the inhibitory effect of the above hesperetin on the in vitro proliferation of three porcine viruses, PEDV, PRRSV, and ASFV, was verified, and the verification method is as follows:

[0056] 1. Materials and methods:

[0057] 1.1 Viruses: PEDV YN strain, PRRSV, and ASFV were isolated, preserved, and donated by the National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University.

[0058] 1.2 Cells: African green monkey kidney cells (Vero) were used for the proliferation of the PEDV YN strain; monkey embryonic kidney epithelial cells (Marc-145) were used for the proliferation of PRRSV; porcine alveolar macrophages (primary cells) were used for the proliferation of ASFV. Vero and Marc-145 cells are cell lines preserved in the laboratory, and porcine alveolar macrophages were isolated from the lungs of healthy piglets.

[0059] 1.3 Test drugs: Hesperetin (provided by Hubei Haohua Biotechnology Co., Ltd.).

[0060] 1.4 Reagents: DMEM medium, EDTA, trypsin, fetal bovine serum, and double antibodies (penicillin-streptomycin) were all purchased from GIBCO.

[0061] 1.5 In vitro proliferation inhibition:

[0062] (1) In vitro proliferation inhibition test of PEDV: All in vitro tests were set up with three experimental groups and one blank group. Different doses of hesperetin were added to the culture medium in the experimental groups. The low-dose hesperetin group (added at 20 μg / mL of the culture medium), the medium-dose hesperetin group (added at 40 μg / mL of the culture medium), the high-dose hesperetin group (added at 80 μg / mL of the culture medium), and the blank group (no hesperetin added to the culture medium). Vero cells were infected with the PEDV DR13 strain (MOI = 0.001), and different doses of hesperetin were added simultaneously during infection. After 36 h of infection, samples were collected, and the expression levels of the M, N, and S genes in the PEDV DR13 strain were detected by qPCR to verify the effect of hesperetin on the in vitro proliferation inhibition of the PEDV DR13 strain.

[0063] (2) In vitro proliferation inhibition test of PRRSV: All in vitro tests were set up with three experimental groups and one blank group. Different doses of hesperetin were added to the culture medium in the experimental groups. The low-dose hesperetin group (added at 20 μg / mL of the culture medium), the medium-dose hesperetin group (added at 40 μg / mL of the culture medium), the high-dose hesperetin group (added at 80 μg / mL of the culture medium), and the blank group (no hesperetin added to the culture medium). Marc-145 cells were infected with the PRRSV strain (MOI = 0.001), and different concentrations of hesperetin were added simultaneously during infection. After 36 h of infection, samples were collected, and the expression levels of the ORF5 and ORF7 genes in the PRRSV strain were detected by qPCR to verify the effect of hesperetin on the in vitro proliferation inhibition of the PRRSV strain.

[0064] (2) In vitro proliferation inhibition test of ASFV: All in vitro tests were set up with three experimental groups and one blank group. Different doses of hesperetin were added to the culture medium in the experimental groups. The low-dose hesperetin group (added at 20 μg / mL of the culture medium), the medium-dose hesperetin group (added at 40 μg / mL of the culture medium), the high-dose hesperetin group (added at 80 μg / mL of the culture medium), and the blank group (no hesperetin added to the culture medium). Porcine alveolar macrophages were infected with the ASFV strain (MOI = 0.001), and different concentrations of hesperetin were added simultaneously during infection. After 36 h of infection, samples were collected, and the expression levels of the P72 and CD2v genes in the ASFV strain were detected by qPCR to verify the effect of hesperetin on the in vitro proliferation inhibition of the PRRSV strain.

[0065] 2. Experimental results:

[0066] The intervention effect of hesperetin on the proliferation of the PEDV strain in Vero cells is shown in Table 2 below, where a, b, and c are signs of significant differences.

[0067] Table 2

[0068] Detected Gene Blank Group 20μg / mL Group 40μg / mL Group 80μg / mL Group M <![CDATA[1.000±0.192 a > <![CDATA[0.517±0.069 b > <![CDATA[0.479±0.055 b > <![CDATA[0.425±0.067 c > N <![CDATA[1.000±0.181 a > <![CDATA[0.521±0.078 b > <![CDATA[0.489±0.079 b > <![CDATA[0.419±0.079 c > S <![CDATA[1.000±0.197 a > <![CDATA[0.531±0.077 b > <![CDATA[0.498±0.089 b > <![CDATA[0.399±0.058 c >

[0069] The results in Table 2 show that adding hesperetin can significantly reduce the expression levels of the N, M, and S genes of PEDV, indicating that hesperetin can significantly reduce the proliferation ability of porcine PEDV.

[0070] The intervention effect of hesperetin on the proliferation of PRRSV strains in IPAM cells is shown in Table 3 below, where a, b, and c are signs of significant differences.

[0071] Table 3

[0072] Detected Gene Blank Group 20μg / mL Group 40μg / mL Group 80μg / mL Group ORF5 <![CDATA[1.000±0.101 a > <![CDATA[0.498±0.053 b > <![CDATA[0.431±0.015 b > <![CDATA[0.313±0.052 c > ORF7 <![CDATA[1.000±0.099 a > <![CDATA[0.502±0.011 b > <![CDATA[0.419±0.053 b > <![CDATA[0.293±0.011 c >

[0073] The results in Table 3 show that adding hesperetin can significantly reduce the expression levels of the ORF5 and ORF7 genes of PRRSV, and the proliferation ability of PRRSV can be significantly reduced after adding hesperetin.

[0074] The intervention effect of hesperetin on the proliferation of ASFV strains in porcine alveolar macrophages is shown in Table 4 below, where a, b, and c are signs of significant differences.

[0075] Table 4

[0076]

[0077]

[0078] The results in Table 4 show that adding hesperetin can significantly reduce the expression levels of the P72 and CD2v genes of ASFV, and the proliferation ability of ASFV can be significantly reduced after adding hesperetin.

[0079] Protective Effect of Hesperetin Preparation on PEDV-infected Piglets

[0080] In the embodiment of the present application, the protective effect of the hesperetin preparation on PEDV-infected piglets was verified, and the verification method is as follows:

[0081] 1. Materials and methods:

[0082] 1.1 Test virus: The PEDV YN strain was kindly provided by the National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University.

[0083] 1.2 Test drug: Hesperetin oral liquid C1.

[0084] 1.3 Experimental animals: Select 7-day-old healthy piglets of the same breed, source, similar body weight, and without maternal antibodies for the challenge test. The experimental piglets are divided into three experimental groups (low, medium, and high-dose groups) and a blank group. Naringenin oral liquid C1 is dissolved in milk and made up to 3 mL for gavage. Among them, the low-dose group (added according to the naringenin content of 2 mg / kg pig body weight), the medium-dose group (added according to the naringenin content of 5 mg / kg pig body weight), the high-dose naringenin group (added according to the naringenin content of 10 mg / kg pig body weight), and the blank group (gavage with the same dose of milk without naringenin preparation). Each group has 10 piglets.

[0085] 1.4 Challenge test: After one week of feeding, the piglets in each group are challenged with the PEDV YN strain at a concentration of 10 7 TCID 50 / mL. Each pig is orally perfused with 5.0 mL. The control group is gavaged with the same dose of cell culture medium (DMEM), and then isolated and raised. Observation is continued for 7 days until the end of the experiment.

[0086] 1.5 Observation indicators: Observe the diarrhea situation of the experimental piglets every day. At the end of the experiment, the experimental piglets are slaughtered to observe intestinal lesions. At the same time, count the number of dead pigs and calculate the diarrhea rate, mortality rate, and protection rate. The inhibitory effect of the naringenin preparation on the PEDV virus is determined by detecting the expression levels of the PEDV structural genes N or M in the intestinal mucosa of piglets by real-time quantitative PCR.

[0087] 1.6 Data analysis: The experimental data are analyzed by one-way ANOVA and Duncan's multiple comparison in the SPSS 13.0 statistical software. P < 0.05 is used as the standard for significant difference, and P < 0.01 is used as the standard for extremely significant difference. The results are expressed as the mean value.

[0088] 2. Experimental results:

[0089] After adding different doses of naringenin oral liquid C1 to milk and feeding it to piglets infected with PEDV, the morbidity and mortality rates of the piglets in each group are shown in Table 5, where a, b, and c are the signs of significant difference.

[0090] Table 5

[0091]

[0092]

[0093] Table 5 results showed that hesperetin oral solution C1 at different doses could significantly reduce the diarrhea rate and mortality of piglets infected with PEDV. Compared with the blank group, the diarrhea rate of piglets infected with PEDV decreased to 10% - 70%; while the mortality rate of diseased pigs decreased to 0 - 50%. The test results fully demonstrated that the addition of hesperetin preparation could significantly improve the protection rate of piglets against PEDV infection.

[0094] The expression levels of the structural genes M, N, and S of PEDV in the intestinal mucosa of the jejunum, ileum, and colon tissues of piglets were detected respectively. The test results are shown in Table 6 below, where a, b, c, and d are the marks for significant differences.

[0095] Table 6

[0096]

[0097] Table 6 results showed that hesperetin oral solution C1 at different doses could significantly reduce the expression levels of the N, M, and S genes of PEDV, indicating that the addition of hesperetin preparation could significantly inhibit the proliferation ability of PEDV virus in the pig intestine.

[0098] Protective Effect of Hesperetin Preparation on PRRSV-infected Piglets

[0099] In the embodiment of this application, the protective effect of hesperetin preparation on piglets infected with PRRSV was verified. The verification method is as follows:

[0100] 1. Materials and methods:

[0101] 1.1 Test virus: The PRRSV strain was kindly provided by the National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University.

[0102] 1.2 Test drug: Hesperetin powder C4.

[0103] 1.3 Test animals: Healthy 25-day-old piglets with the same breed, source, similar body weight, and no maternal antibodies were selected for the challenge test. The test piglets were divided into three test groups (low, medium, and high dose groups) and a blank group. Hesperetin was dissolved in milk and made up to 3 mL for gavage. Among them, the low dose group (added according to the hesperetin content of 2 mg / kg body weight of pigs), the medium dose group (added according to the hesperetin content of 5 mg / kg body weight of pigs), the high dose group (added according to the hesperetin content of 10 mg / kg body weight of pigs), and the blank group (fed with the same dose of milk without adding hesperetin preparation), with 10 piglets in each group.

[0104] 1.4 Challenge test: After the piglets in each group were fed for 1 week, at a concentration of 10 7 TCID 50Pigs were challenged with 10⁶ TCID₅₀ / mL of PRRSV strain by oral perfusion of 5 mL per pig. The control group was given the same dose of normal saline by gavage, and then the pigs were isolated and observed continuously for 7 days until the end of the experiment.

[0105] 1.5 Observation indicators: The status of the experimental piglets was observed daily. At the end of the experiment, the piglets were slaughtered to observe the lung lesions, and at the same time, the number of dead pigs was counted, and the morbidity and mortality rates were calculated. The inhibitory effect of hesperetin on PRRSV was determined by detecting the expression level of PRRSV ORF7 gene in the brains and lungs of piglets by real-time quantitative PCR.

[0106] 1.6 Data analysis: The experimental data were analyzed by one-way ANOVA and Duncan's multiple comparison in SPSS 13.0 statistical software. The significance level was set at P < 0.05 for significant differences and P < 0.01 for extremely significant differences. The results were expressed as mean values.

[0107] 2. Experimental results:

[0108] After adding different doses of hesperetin powder C4 to milk and feeding it to piglets infected with PRRSV, the morbidity and mortality rates of each group of piglets are shown in Table 7, where a, b, and c are the signs of significant differences.

[0109] Table 7

[0110] Group Incidence Rate(%) Mortality Rate(%) Protection Rate(%) Blank Group <![CDATA[100 a > <![CDATA[80 b > <![CDATA[20 c > 2mg / kg Group <![CDATA[60 a > <![CDATA[40 b > <![CDATA[60 a > 5mg / kg Group <![CDATA[30 b > <![CDATA[10 c > <![CDATA[90 a <!-- 7 -->]]> 10mg / kg Group <![CDATA[20 b > <![CDATA[10 c > <![CDATA[90 a >

[0111] The results in Table 7 show that oral administration of different doses of hesperetin powder C4 to piglets can significantly reduce the morbidity and mortality rates of PRRSV-infected piglets. Compared with the blank group, as the dose increases, the morbidity rate of PRRSV-infected piglets decreases to 20% - 60%; while the mortality rate of diseased pigs decreases to 10% - 40%. The experimental results fully show that the addition of hesperetin preparation can significantly improve the protection rate of piglets against PRRSV infection.

[0112] The expression levels of the PRRSV structural gene ORF7 in the brains and lungs of piglets were detected respectively, and the detection results are shown in Table 8 below, where a, b, c, and d are the signs of significant differences.

[0113] Table 8

[0114]

[0115] The results in Table 8 show that adding different doses of hesperetin powder C1 can significantly reduce the expression level of the PRRSV ORF7 gene. The results fully show that the addition of hesperetin preparation can significantly reduce the replication of PRRSV in the brains and lungs.

[0116] In summary, the hesperetin preparation provided by the embodiments of the present application has the effect of resisting porcine enveloped viruses such as PEDV, PRRSV, and ASFV, and can be used to prepare drugs and feed additives for treating and preventing porcine diseases caused by porcine enveloped virus infections such as PEDV, PRRSV, and ASFV. The dosage forms of the drugs and feed additives include oral liquids, granules, powders, tablets and other dosage forms.

[0117] The above has introduced the present application in detail. Specific examples are used in this article to elaborate on the principle and implementation manner of the present application. The description of the above embodiments is only used to help understand the present application and its core idea. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present application, several improvements and modifications can be made to the present application, and these improvements and modifications also fall within the protection scope of the claims of the present application.

Claims

1. Use of hesperetin in the preparation of a medicament or feed additive for treating or preventing porcine enveloped virus infection, wherein the porcine enveloped virus includes at least one of porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, and African swine fever virus.

2. The application according to claim 1, wherein The medicament or feed additive is a composition comprising hesperetin and excipients, and the excipients include at least one of absolute ethanol, water, Tween-80, lactose, defatted rice bran, corn cob powder, soluble starch, wheat bran, silica, montmorillonite, vermiculite, medical stone, and zeolite powder.

3. The application according to claim 2, wherein The composition comprising hesperetin and excipients is a solution formed by mixing hesperetin with three solvents of absolute ethanol, Tween-80, and water, wherein the concentration of hesperetin is 20 - 100 mg / mL.

4. The application according to claim 2, characterized in that, The composition comprising hesperetin and excipients is a solid mixture formed by mixing hesperetin with lactose and soluble starch, wherein the mass fraction of hesperetin in the solid mixture is 45% - 55%.

5. The application according to claim 2, wherein The aforementioned composition comprising hesperetin and excipients is a powdery mixture formed by mixing hesperetin with soluble starch or wheat bran, wherein the mass fraction of hesperetin in the powdery mixture is 45% - 55%.

6. The application according to claim 2, wherein When the medicament or feed additive is a composition comprising hesperetin and excipients, its preparation types include at least one of oral liquid, granule, tablet, and powder.

7. The application according to claim 1, wherein The usage methods of the medicament or feed additive include at least one of (1) - (3): (1) Add hesperetin or the composition comprising hesperetin and excipients to feed or drinking water for use at a dose of 2 - 10 mg / kg of pig body weight based on the content of hesperetin; (2) Administer hesperetin or the composition comprising hesperetin and excipients directly by oral administration at a dose of 2 - 10 mg / kg of pig body weight based on the content of hesperetin; (3) Add hesperetin or the composition comprising hesperetin and excipients to feed for use at a dose of 0.01% - 0.03% of the total weight of pig feed based on the content of hesperetin.

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