Application of hesperetin in preparation of medicine or feed additive for treating or preventing swine envelope virus infection

Abstract

The invention discloses an application of hesperetin in preparation of a medicine or a feed additive for treating or preventing swine envelope virus infection, the medicine is hesperetin or is prepared into oral liquid, granules, tablets and powder by taking hesperetin as an effective component and adding a solvent or an auxiliary material, and the use dosage is 2-10mg / kg of the body weight of a pig; or the hesperetin is directly added into the feed, and the addition amount of the hesperetin is 0.01-0.03% of the total weight of the pig feed. The hesperetin or the composition containing the hesperetin is used as a medicine or a feed additive, is high in safety and free of toxic and side effects, and has the effects of remarkably inhibiting replication of envelope viruses such as porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus and African swine fever virus and reducing the toxicity of the porcine envelope viruses; the defects of immune failure possibly caused by vaccination, virus variation caused by use of antiviral drugs and the like are overcome, and the morbidity and mortality of the piglets infected with the swine envelope virus infection are effectively reduced.

Description

Technical Field

[0001] The present application relates to the technical field of prevention and treatment of porcine envelope virus, and specifically to the application of hesperidin in the preparation of medicines or feed additives for treating or preventing porcine envelope virus infection. Background Art

[0002] Porcine enveloped viruses refer to a class of swine viruses with an envelope structure, which can cause a variety of 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 bind to receptors, enter cells to replicate and proliferate, and antagonize the host's natural immune response, causing immunosuppression. PEDV mainly causes vomiting and watery diarrhea in newborn piglets within 10 days of age, and can lead to high morbidity and mortality in newborn piglets. PRRSV infection can cause porcine reproductive and respiratory syndrome (PRRS). The virus mainly infects pig alveolar macrophages, leading to immune system dysfunction, which is mainly manifested by low and short-lasting neutralizing antibody response levels; cellular immune function is suppressed, and the expression level of interferon-γ is low and slow; infected pigs continue to carry the virus, and the virus is difficult to eliminate from the pig herd. ASFV is the pathogen that causes African swine fever (ASF). ASF is a highly pathogenic, high-mortality, and highly contagious animal epidemic disease. Its most acute mortality rate is as high as 100%. It is one of the major animal infectious diseases that must be reported 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 farming industry. At present, the prevention and control measures for PEDV, PRRSV, and ASFV viral infections in my country mainly rely on vaccines. However, due to the abuse of large doses of antiviral and antibacterial drugs, the mutation rate of the virus (bacteria) strains far exceeds the speed of vaccine development, and the immunosuppressive characteristics of the virus lead to poor vaccine immunity. In addition, since vaccine immunity only has the function of preventing the occurrence of viral diseases, it cannot provide protection for piglets that have already been infected and become ill. Therefore, relying solely on vaccines can easily lead to immune failure, and large-scale PEDV, PRRSV, and ASFV viral infections still often break out.

[0004] In view of the problems of existing vaccines in preventing and treating porcine envelope virus, the development of functional nutrients with resistance to porcine envelope virus will have broad application prospects. Summary of the invention

[0005] In view of this, the purpose of the present application is to provide an application of hesperidin in the preparation of a drug or feed additive for treating or preventing porcine envelope virus infection. The hesperidin is a natural additive that has the effect of resisting porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus and African swine fever virus, has high safety and is easy to use. The hesperidin can be used to prepare a drug or feed additive for treating or preventing porcine envelope virus infection.

[0006] In order to achieve or at least partially achieve the above objectives, the present application provides the following technical solutions:

[0007] Application of hesperidin in preparing medicine or feed additive for treating or preventing porcine envelope virus infection.

[0008] Compared with the prior art, the technical solution provided by this application has at least the following beneficial effects:

[0009] 1. The hesperidin or preparation containing hesperidin provided in the present application can be used as a feed additive for a long time to achieve the purpose of preventing epidemic diseases. Long-term use will not lead to the production of drug-resistant strains, nor will it have other toxic side effects, overcoming the shortcomings of possible immune failure caused by vaccination and virus mutation caused by the use of antiviral drugs. After feeding the hesperidin or preparation containing hesperidin to pigs, it can enhance the body's antioxidant defense function, improve the pig's natural immune function and antiviral activity, and effectively prevent and treat infections of porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus and African swine fever virus.

[0010] 2. This application proves through in vitro experimental results that hesperidin has a significant effect of inhibiting the proliferation of viruses such as PEDV, PRRSV and ASFV.

[0011] 3. The results of in vivo experiments in this application show that hesperidin has the ability to enhance natural immune function, antiviral activity, improve pig intestinal structure and absorption function, effectively prevent and treat infection with porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, and African swine fever virus, and effectively improve the survival rate and production performance of pigs. Oral administration of 10 mg / kg pig body weight or adding hesperidin to piglet feed can achieve a protection rate of more than 90% against infection with porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, and African swine fever virus. DETAILED DESCRIPTION

[0012] In order to make the purpose, technical solution and advantages of the present application more clearly understood, the present application is further described in detail below in combination with the 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 will appreciate that, unless otherwise stated, the words "said", "the", and "the aforementioned" used in the text of the present application may also include plural forms. It should be further understood that the wording "including" used in the specification of the present application means the presence of the features, steps, and operations, but does not exclude the presence or addition of one or more other features, integers, and steps.

[0014] Those skilled in the art will appreciate that, for any specific experimental steps or conditions not specified in the examples, the experiments may be carried out according to the conventional experimental steps or conditions described in the literature in the art; any raw materials or instruments used without specifying the manufacturer are conventional products that can be purchased commercially.

[0015] Those skilled in the art will understand that, unless otherwise specified in the present application, when a numerical range is given in an embodiment, both 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 this application are consistent with the prior art mastery of those skilled in the art and the records of this application, and any methods, equipment and materials of the prior art similar or equivalent to the methods, equipment and materials described in the embodiments of this application can also be used to implement this application.

[0016] Hesperetin (HES) is a well-known and common flavonoid glycoside in Chinese medicine (TCM). 16 H 14 O6, usually appears as light yellow needle-shaped crystals or powder, and some say it is beige to light brown crystalline solid. HES is present in high levels in Citrus aurantium, Citrus aurantium immaturus, and dried orange peel, and is mostly present in Rutaceae plants in the form of glycosides.

[0017] The examples of the present application found through in vivo and in vitro experiments that hesperidin and a composition containing hesperidin can significantly inhibit the proliferation of PEDV, PRRSV and ASFV, and inhibit their infection of cells. The hesperidin and a composition containing hesperidin (hereinafter referred to as "hesperidin 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 use of hesperidin in the preparation of a drug or feed additive for treating or preventing porcine envelope virus infection.

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

[0020] In certain embodiments, the drug or feed additive is hesperetin, which can be directly used as a drug or feed additive to treat or prevent pig diseases such as PED, PRRS and ASF.

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

[0022] In certain embodiments, the aforementioned hesperetin composition is a solution of hesperetin mixed with three solvents of anhydrous ethanol, Tween-80, and water, wherein the concentration of hesperetin is 20-100 mg / mL.

[0023] In certain embodiments, the aforementioned hesperetin composition is a solid mixture of hesperetin, lactose and soluble starch, wherein the mass fraction of hesperetin in the solid mixture is 45% to 55%.

[0024] In certain embodiments, the aforementioned hesperetin composition is a powdery mixture of hesperetin and soluble starch or bran, wherein the mass fraction of hesperetin in the powdery mixture is 45% to 55%.

[0025] In certain embodiments, when the aforementioned medicine or feed additive is a hesperidin composition, its preparation type includes at least one of oral liquid, granules, tablets, and powders, and the preparation can be prepared by conventional pharmaceutical preparation methods.

[0026] In certain embodiments, the method of using the drug or feed additive comprises at least one of the following methods:

[0027] (1) adding hesperidin or a hesperidin composition to feed or drinking water at a dosage of 2 to 10 mg / kg of pig body weight;

[0028] (2) directly orally administering hesperetin or a hesperetin composition at a dosage of 2 to 10 mg / kg of pig body weight;

[0029] (3) Hesperidin or the hesperidin composition is added to the feed at a dosage of 0.01% to 0.03% of the total weight of the pig feed.

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

[0031] Example 1

[0032] This embodiment provides a method for preparing an anti-swine envelope virus hesperidin oral liquid, the specific steps comprising:

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

[0034] (2) First, hesperidin was dissolved in anhydrous ethanol, then Tween-80 was added, and finally water for injection was slowly added to make up to 100 mL; the mixture was mixed evenly to obtain a 20 mg / mL hesperidin solution, i.e., the hesperidin oral solution C1.

[0035] Example 2

[0036] This embodiment provides a method for preparing an anti-swine envelope virus hesperidin oral liquid, the specific steps comprising:

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

[0038] (2) First, hesperidin was dissolved in anhydrous ethanol, then Tween-80 was added, and finally water for injection was slowly added to make up to 100 mL; the mixture was mixed evenly to obtain a 100 mg / mL hesperidin solution, i.e., the hesperidin oral solution C2.

[0039] Example 3

[0040] This embodiment provides a method for preparing hesperidin granules for resisting porcine envelope virus, and the specific steps include:

[0041] (1) Weigh 50 g of hesperidin, 25 g of lactose, and 25 g of soluble starch and set aside;

[0042] (2) Hesperidin, lactose and soluble starch are mixed and granulated, and the granulated mixture is dried to obtain a granule having a hesperidin mass fraction of 50%, namely, hesperidin granule C3.

[0043] Example 4

[0044] This embodiment provides a method for preparing hesperidin powder for resisting porcine envelope virus, and the specific steps include:

[0045] (1) Weigh 50 g of hesperidin and 50 g of bran and set aside;

[0046] (2) The hesperidin and bran were mixed evenly and passed through a 60-mesh sieve to obtain a powdery mixture with a hesperidin mass fraction of 50%, i.e., hesperidin powder C4.

[0047] Example 5

[0048] This embodiment provides a method for preparing an anti-swine endotoxin virus hesperidin tablet, the specific steps comprising:

[0049] (1) Weigh 50 g of hesperidin, 25 g of lactose, and 25 g of soluble starch and set aside;

[0050] (2) After the hesperidin, lactose and soluble starch are uniformly mixed, the mixture is compressed and dried using a tablet press to obtain a flaky mixture having a mass fraction of hesperidin of 50%, namely the hesperidin tablet C5.

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

[0052] Table 1

[0053] Example No. Preparation No. Formulation type Components Terminalia chebula tannin chelate concentration Example 1 C1 Oral solution Hesperidin, anhydrous ethanol, Tween-80 20mg / mL Example 2 C2 Oral solution Hesperidin, anhydrous ethanol, Tween-80 100mg / mL Example 3 C3 Granules Hesperidin, lactose, soluble starch 50% (mass fraction) Example 4 C4 powder Hesperidin, bran 50% (mass fraction) Example 5 C5 tablet Hesperidin, lactose, soluble starch 50% (mass fraction)

[0054] Inhibitory effect of hesperidin on proliferation of PEDV, PRRSV and ASFV in vitro

[0055] In the present application, the inhibitory effect of hesperidin on the proliferation of three porcine viruses, PEDV, PRRSV and ASFV, was verified by the following method:

[0056] 1. Materials and methods:

[0057] 1.1 Viruses: PEDVYN 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) are used for the proliferation of PEDVYN strains; monkey embryonic kidney epithelial cells (Marc-145) are used for the proliferation of PRRSV; porcine alveolar macrophages (primary cells) are used for the proliferation of ASFV. Vero and Marc-145 cells are laboratory preserved cell lines, and porcine alveolar macrophages are isolated from the lungs of healthy piglets.

[0059] 1.3 Experimental drug: Hesperidin (provided by Hubei Haohua Biotechnology Co., Ltd.).

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

[0061] 1.5 In vitro proliferation inhibition:

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

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

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

[0065] 2. Experimental results:

[0066] The intervention effect of hesperidin on the proliferation of PEDV strains in Vero cells is shown in Table 1, where a, b, and c are significant differences.

[0067] Table 1

[0068] Detection of genes Blank Group 20 μg / mL group 40 μg / mL group 80 μg / 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 1 show that the addition of hesperetin can significantly reduce the expression levels of N, M and S genes of PEDV, indicating that hesperetin can significantly reduce the proliferation ability of porcine PEDV.

[0070] The intervention effect of hesperidin on the proliferation of PRRSV strains in IPAM cells is shown in Table 2 below, where a, b, and c are significant differences.

[0071] Table 2

[0072] Detection of genes Blank Group 20 μg / mL group 40 μg / mL group 80 μg / 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 2 show that the addition of hesperidin can significantly reduce the expression levels of ORF5 and ORF7 genes of PRRSV, and the addition of hesperidin can significantly reduce the proliferation ability of PRRSV.

[0074] The intervention effect of hesperidin on the proliferation of ASFV strains in pig alveolar macrophages is shown in Table 3, where a, b, and c are significant differences.

[0075] Table 3

[0076] Detection of genes Blank Group 20 μg / mL group 40 μg / mL group 80 μg / mL group P72 1.000±0.201a 0.559±0.076b 0.463±0.051c 0.391±0.063d CD2v 1.000±0.197a 0.621±0.083b 0.498±0.073c 0.413±0.087d

[0077] The results in Table 3 show that the addition of hesperidin can significantly reduce the expression levels of P72 and CD2v genes of ASFV, and the addition of hesperidin can significantly reduce the proliferation ability of ASFV.

[0078] Protective effect of hesperetin preparation on piglets infected with PEDV

[0079] The present application example verifies the protective effect of hesperidin preparation on piglets infected with PEDV, and the verification method is as follows:

[0080] 1. Materials and methods:

[0081] 1.1 Experimental virus: PEDVYN strain was donated by the National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University.

[0082] 1.2 Experimental drug: Hesperidin oral solution C1.

[0083] 1.3 Experimental animals: 7-day-old healthy piglets of the same breed and origin, similar weight and without maternal antibodies were selected for the challenge test. The experimental piglets were divided into three experimental groups (low, medium and high dose groups) and a blank group. Hesperidin oral solution C1 was dissolved in milk and the volume was made up to 3 mL for oral administration, including the low-dose group (addition of hesperidin content of 2 mg / kg pig body weight), the medium-dose group (addition of hesperidin content of 5 mg / kg pig body weight), the high-dose group (addition of hesperidin content of 10 mg / kg pig body weight) and the blank group (gavage feeding of the same dose of milk without hesperidin preparation), each group had 10 piglets.

[0084] 1.4 Challenge test: After feeding for one week, the piglets in each group were fed with 10 7 TCID 50 / mL of PEDVYN strain was used for infection, and 5.0mL was orally infused to each head. The control group was infused with the same dose of cell culture medium (DMEM), and then they were isolated and raised and observed for 7 consecutive days until the end of the experiment.

[0085] 1.5 Observation indicators: The diarrhea of ​​the experimental piglets was observed every day. The experimental piglets were slaughtered at the end of the experiment to observe intestinal lesions. The number of dead pigs was counted and the diarrhea rate, mortality rate and protection rate were calculated. The inhibitory effect of hesperidin preparation on PEDV virus was determined by detecting the expression of PEDV structural gene N or M in the intestinal mucosa of piglets through real-time quantitative PCR.

[0086] 1.6 Data analysis: The experimental data were analyzed using one-way ANOVA and Duncan's multiple comparison method in SPSS13.0 statistical software, with P < 0.05 as the standard of significant difference and P < 0.01 as the standard of extremely significant difference. The results are expressed as mean values.

[0087] 2. Experimental results:

[0088] After adding different doses of hesperidin oral solution C1 to milk and feeding it to piglets infected with PEDV, the morbidity and mortality of piglets in each group are shown in Table 4, where a, b, and c are significant differences.

[0089] Table 4

[0090]

[0091]

[0092] The results in Table 4 show that different doses of hesperidin oral solution C1 can significantly reduce the diarrhea rate and mortality rate of PEDV-infected piglets. Compared with the blank group, the diarrhea rate of PEDV-infected piglets decreased to 10%-70%; and the mortality rate of diseased pigs decreased to 0-50%. The experimental results fully demonstrate that the addition of hesperidin preparations can significantly improve the protection rate of piglets against PEDV infection.

[0093] The expression levels of PEDV structural genes M, N, and S in the intestinal mucosa of the jejunum, ileum, and colon tissues of piglets were detected respectively. The test results are shown in Table 5 below, where a, b, c, and d are significant differences.

[0094] Table 5

[0095]

[0096] The results in Table 5 show that different doses of hesperidin oral solution C1 can significantly reduce the expression levels of N, M and S genes of PEDV, indicating that the addition of hesperidin preparation can significantly inhibit the proliferation ability of PEDV virus in pig intestine.

[0097] Protective effect of hesperetin preparation on piglets infected with PRRSV

[0098] The present application example verifies the protective effect of hesperidin preparation on piglets infected with PRRSV, and the verification method is as follows:

[0099] 1. Materials and methods:

[0100] 1.1 Experimental virus: The PRRSV strain was donated by the National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University.

[0101] 1.2 Experimental drug: Hesperidin powder C4.

[0102] 1.3 Experimental animals: 25-day-old healthy piglets of the same breed and origin, similar weight and without maternal antibodies were selected for the challenge test. The experimental piglets were divided into three experimental groups (low, medium and high dose groups) and a blank group. Hesperidin was dissolved in milk and the volume was made up to 3 mL for oral administration, including the low-dose group (addition of hesperidin content of 2 mg / kg pig body weight), the medium-dose group (addition of hesperidin content of 5 mg / kg pig body weight), the high-dose group (addition of hesperidin content of 10 mg / kg pig body weight) and the blank group (gavage feeding of the same dose of milk without hesperidin preparation), with 10 piglets in each group.

[0103] 1.4 Challenge test: After feeding for 1 week, the piglets in each group were fed with 10 7 TCID 50 / mL PRRSV strain was used for infection, and 5mL was orally infused into each head. The control group was infused with the same dose of normal saline, and then they were isolated and raised, and observed for 7 consecutive days until the end of the experiment.

[0104] 1.5 Observation indicators: The status of the experimental piglets was observed every day. The experimental piglets were slaughtered at the end of the experiment to observe lung lesions, and the number of dead pigs was counted, and the incidence and mortality rates were calculated. The inhibitory effect of hesperidin on PRRSV was determined by detecting the expression of PRRSVORF7 gene in the brain and lungs of piglets through real-time quantitative PCR.

[0105] 1.6 Data analysis: The experimental data were analyzed using one-way ANOVA and Duncan's multiple comparison method in SPSS13.0 statistical software, with P < 0.05 as the standard of significant difference and P < 0.01 as the standard of extremely significant difference. The results are expressed as mean values.

[0106] 2. Experimental results:

[0107] After adding different doses of hesperidin powder C4 to milk and feeding it to piglets infected with PRRSV, the morbidity and mortality of piglets in each group are shown in Table 6, where a, b, and c are significant differences.

[0108] Table 6

[0109] Group Incidence (%) 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 ]]>

[0110] The results in Table 6 show that oral administration of different doses of hesperidin powder C4 to piglets can significantly reduce the morbidity and mortality of piglets infected with PRRSV. Compared with the blank group, with the increase of dose, the morbidity of piglets infected with PRRSV decreased to 20%-60%; and the mortality of diseased pigs decreased to 10%-40%. The experimental results fully show that the addition of hesperidin preparation can significantly improve the protection rate of piglets against PRRSV infection.

[0111] The expression levels of PRRSV structural gene ORF7 in the brain and lung of piglets were detected respectively. The detection results are shown in Table 7 below, where a, b, c, and d are marks of significant difference.

[0112] Table 7

[0113]

[0114] The results in Table 7 show that adding different doses of hesperidin powder C1 can significantly reduce the expression of ORF7 gene of PRRSV. The results fully demonstrate that the addition of hesperidin preparation can significantly reduce the replication of PRRSV in the brain and lungs.

[0115] In summary, the hesperidin preparation provided in the embodiments of the present application has the effect of resisting porcine envelope viruses such as PEDV, PRRSV and ASFV, and can be used to prepare drugs and feed additives for treating and preventing pig diseases infected by porcine envelope viruses such as PEDV, PRRSV and ASFV. The dosage forms of the drugs and feed additives include oral liquids, granules, powders, tablets and the like.

[0116] The present application is described in detail above. Specific examples are used in this article to illustrate the principles and implementation methods of the present application. The description of the above embodiments is only used to help understand the present application and its core ideas. It should be pointed out that for ordinary technicians in this technical field, without departing from the principles of the present application, several improvements and modifications can be made to the present application, and these improvements and modifications also fall within the scope of protection of the claims of the present application.

Claims

1. The application of hesperidin in the preparation of medicines or feed additives for treating or preventing porcine envelope virus infection.

2. The use according to claim 1, characterized in that: The porcine virus includes at least one of porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus, and African swine fever virus.

3. The use according to claim 1, characterized in that: The medicine or feed additive is hesperetin.

4. The use according to claim 1, characterized in that: The medicine or feed additive is a composition comprising hesperidin and auxiliary materials, wherein the auxiliary materials comprise at least one of anhydrous ethanol, water, Tween-80, lactose, defatted rice bran, corn cob powder, soluble starch, bran, silicon dioxide, montmorillonite, vermiculite, medical stone, zeolite powder and white carbon black.

5. The use according to claim 4, characterized in that: The hesperidin composition is a solution formed by mixing hesperidin with three solvents: anhydrous ethanol, Tween-80 and water, wherein the concentration of hesperidin is 20-100 mg / mL.

6. The use according to claim 4, characterized in that: The hesperidin composition is a solid mixture of hesperidin, lactose and soluble starch, wherein the mass fraction of hesperidin in the solid mixture is 45% to 55%.

7. The use according to claim 4, characterized in that: The aforementioned hesperetin composition is a powdery mixture of hesperetin and soluble starch or bran, wherein the mass fraction of hesperetin in the powdery mixture is 45% to 55%.

8. The use according to claim 4, characterized in that: When the medicine or feed additive is a composition comprising hesperidin and auxiliary materials, its preparation type includes at least one of oral liquid, granules, tablets and powders.

9. The use according to claim 1, characterized in that: The method for using the drug or feed additive includes at least one of (1) to (3): (1) adding hesperidin or a hesperidin composition to feed or drinking water at a dosage of 2 to 10 mg / kg of pig body weight; (2) directly orally administering hesperetin or a hesperetin composition at a dosage of 2 to 10 mg / kg of pig body weight; (3) Hesperidin or the hesperidin composition is added to the feed at a dosage of 0.01% to 0.03% of the total weight of the pig feed.

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