CN112741899A

CN112741899A - Composition, food supplement and application of composition and food supplement in inhibiting animal herpesvirus

Info

Publication number: CN112741899A
Application number: CN202011637366.1A
Authority: CN
Inventors: 单萌; 孙雨薇; 李靖
Original assignee: Nanjing Aurora Biotechnology Co ltd
Current assignee: Nanjing Aurora Biotechnology Co ltd
Priority date: 2020-12-31
Filing date: 2020-12-31
Publication date: 2021-05-04

Abstract

According to the composition, the food supplement and the application, lactoferrin and beta-glucan are used as main functional components, so that the composition can play a good role in preventing and treating animal herpesviruses; lactoferrin can be loaded on beta-glucan, successfully enters intestinal tracts, is combined with lactoferrin receptors on intestinal mucosa, and passes through cell transmembrane to enter a circulatory system through receptor-mediated endocytosis to directly reach focuses; the beta-glucan has the function of improving immunity, and plays a role in adjuvant therapy by improving the immunity of the organism in the treatment process. The invention also provides application of the composition and the supplement in the preparation of medicaments, feeds or feed additives for preventing or treating animal herpesvirus, and through in vitro cell tests and animal tests, the supplement provided by the invention has good preventing and treating effects on FHV-1 virus.

Description

Composition, food supplement and application of composition and food supplement in inhibiting animal herpesvirus

Technical Field

The invention relates to a composition, a food supplement and application, and belongs to the technical field of food prevention and treatment of animal herpesviruses.

Background

Herpes viruses (herpesviruses) are a group of enveloped DNA viruses with similar biological properties, classified in the herpesviridae family. It is widely distributed in vertebrates, including fish, amphibians, reptiles, mammals and birds. To date, herpes viruses have also been found in invertebrates such as mollusks. Herpes virus infects a wide range of hosts, mainly affecting the skin, mucous membranes and nervous tissues, and seriously affecting the health of the infected organism.

For example, Feline Viral Rhinotracheitis (FVR), a popular name of Feline Rhinotracheitis, is a virulent infectious disease caused by herpes virus type I (FHV-I), and the main infection site is the upper respiratory tract or lung of a cat. The susceptible groups are cats in all ages, wherein the infection risk of the kitten is higher, and the risks of pregnant cats and cats with low immunity are also higher. The most prominent and effective way to prevent this disease is currently vaccination against FVR, but FVR vaccines do not completely prevent infection when cats are exposed to the virus.

Feline herpes viruses proliferate reproducibly in conjunctival and upper respiratory epithelial cells, and also reproducibly in neuronal cells, with neuronal infections resulting in lifelong latent infections, classified as acute and chronic latent infections, typical of feline herpes viruses after acute infection, and which reactivate intermittently, allowing viral discharge in oronasal and conjunctival secretions. It is clinically manifested as conjunctivitis and keratitis, including conjunctival congestion and conjunctivitis combination, serous fluid to purulent eye secretions, blepharospasm, and few conjunctival ulcers.

At present, the treatment mode is mainly supportive therapy, and the dehydration, the ion correction and the acid-base balance of the affected livestock are corrected through intravenous infusion. Common treatment regimens are: l-lysine (L-lysine) 500mg/cat (kitten 250mg) was used as a supplement for FHV-1 virus management. However, current treatment and prevention regimens have the following drawbacks:

1. supplementation with L-lysine was not effective in preventing infection of the nasal branches of cats;

2. because the kitten cannot synthesize arginine, the supplementation of a large dose of L-lysine may cause the serious inhibition of arginine absorption, so that the liver cannot generate enough enzyme to remove byproducts generated after the digestion and decomposition of protein in food, such as toxic ammonia, and the high concentration of ammonia in the body causes hyperammonemia, which is fatal and has great toxic and side effects;

3. the supplement dosage is larger, so that the treatment cost is increased, and the side effect on the body of the cat is larger.

Lactoferrin, an 80kDa iron-binding glycoprotein, belongs to the transferrin family. Lactoferrin is high in colostrum and milk, and low in tear, saliva, semen, nasal and bronchial secretions, bile and gastrointestinal fluids, etc. In addition, lactoferrin is also a component of neutrophils. Lactoferrin not only participates in the transport of iron, but also has the powerful biological functions of broad-spectrum antibiosis, antioxidation, anticancer, immune system regulation and the like. Researches show that the lactoferrin LF is a natural protein in animal colostrum, is a multifunctional protein, has the functions of broad-spectrum antibiosis and virus infection resistance, and can regulate the balance of iron in a body; regulating the generation of bone marrow cells and promoting the growth of the cells; regulating immunity, and enhancing disease resistance; inhibiting human tumor cells; can be used for treating diseases effectively under the synergistic effect with various antibiotics and antifungal agents. However, the current research on lactoferrin antiviral shows that lactoferrin is degraded into amino acids or small peptides after entering the body and being digested and absorbed by gastrointestinal tracts, so that the application of lactoferrin in antiviral therapy is limited.

Disclosure of Invention

The invention aims to solve the defects and shortcomings in the prior art and provides a supplement for preventing or treating feline rhinotracheitis.

In order to solve the above technical problems, the present invention provides a composition, which is characterized in that: including lactoferrin and beta-glucans;

the lactoferrin class includes but is not limited to one or more of lactoferrin, lactoferricin derivatives, modified products of lactoferricin hydrolysate; the lactoferrin source includes, but is not limited to, mammalian source, and artificial lactoferrin obtained by means of biological fermentation.

The beta-glucan is one or more of beta-glucan and beta-glucan derivatives.

Further, the modified products of the lactoferricin, the lactoferricin derivative and the lactoferricin hydrolysate comprise at least one tryptophan; wherein, the lactoferricin is a section of polypeptide released by the lactoferrin acting on the N end through pepsin in an acidic environment; the lactoferrin peptide derivative and the lactoferrin hydrolysate are obtained by substituting certain amino acid on the lactoferrin peptide or the lactoferrin hydrolysate after the lactoferrin is hydrolyzed by other amino acid so as to change the activity of the lactoferrin peptide or the lactoferrin hydrolysate; in addition, Trp is a hydrophobic amino acid with amino acid side chain, which contains an indole ring and can form a dipole, a quadrupole and an unbound H +, thereby having high antiviral activity.

And/or

The beta-glucan is derived from cereals or microorganisms; cereals including but not limited to barley, oat, highland barley, wheat, rye in one or more mixture, microorganisms including but not limited to yeast, bacteria, fungi in one or more mixture;

and/or

In the beta-glucan and the beta-glucan derivative, the ratio of the content of (1, 3) glycosidic bonds to the content of (1, 4) glycosidic bonds is 1: 1 to (1-4.2), wherein the ratio of the content of (1, 3) glycosidic bonds to the content of (1, 4) glycosidic bonds is preferably 1: 2.4-2.6, and more preferably the ratio of the content of (1, 3) glycosidic bonds to the content of (1, 4) glycosidic bonds is 1: 2.5; it has been found through extensive experiments by the applicant that the ratio of the content of (1, 3) glycosidic linkages to the content of (1, 4) glycosidic linkages is within the above range, and that the water solubility is the best.

And/or

The beta-glucan derivative comprises carboxymethyl glucan, sulfated glucan and beta-glucan phosphate.

Further, the content of lactoferrin is 0.1-200mg/g, and the content of beta-glucan is 10-800 mg/g; preferably, the content of lactoferrin is 5-180mg/g, and the content of beta-glucan is 10-600 mg/g; preferably, the content of lactoferrin is 20-150mg/g, and the content of beta-glucan is 50-550 mg/g; more preferably, the content of lactoferrin is 100mg/g, and the content of β -glucan is 250 mg/g.

The invention also provides a food supplement for animals, which comprises lactoferrin, beta-glucan and auxiliary materials, wherein the lactoferrin comprises one or more of lactoferrin, lactoferricin derivatives, modified products of lactoferricin hydrolysate and lactoferrin obtained by biological fermentation; the beta-glucan is one or more of beta-glucan and beta-glucan derivatives.

Further, the modified products of the lactoferricin, the lactoferricin derivative and the lactoferricin hydrolysate comprise at least one tryptophan; wherein, the lactoferricin is a section of polypeptide released by the lactoferrin acting on the N end through pepsin in an acidic environment; the lactoferrin peptide derivative and the lactoferrin hydrolysate are obtained by substituting certain amino acid on the lactoferrin peptide or the lactoferrin hydrolysate after the lactoferrin is hydrolyzed by other amino acid so as to change the activity of the lactoferrin peptide or the lactoferrin hydrolysate; in addition, Trp is a hydrophobic amino acid with an amino acid side chain, and the side chain contains an indole ring, so that a dipole, a quadrupole and an unbound H + can be formed, and therefore, Trp has high antiviral activity; modification of lactoferrin hydrolysate is not limited to the means of modification as long as the product is obtained, such as: protein-like reaction, deamidation and the like;

and/or

The beta-glucan is derived from cereals or microorganisms; the cereals are preferably one or more of barley, oat, highland barley, wheat and rye, and the microorganisms are preferably one or more of yeast, bacteria and fungi;

and/or

In the beta-glucan and the beta-glucan derivative, the ratio of the content of (1, 3) glycosidic bonds to the content of (1, 4) glycosidic bonds is 1: 1 to (1-4.2), wherein the ratio of the content of (1, 3) glycosidic bonds to the content of (1, 4) glycosidic bonds is preferably 1: 2.4-2.6, and more preferably the ratio of the content of (1, 3) glycosidic bonds to the content of (1, 4) glycosidic bonds is 1: 2.5;

and/or

The beta-glucan derivative comprises carboxymethyl glucan, sulfated glucan and beta-glucan phosphate;

and/or

The auxiliary materials comprise purified water, normal saline, monosaccharides and polysaccharides, protein powder, probiotics, prebiotics, dietary fibers and flavor regulator.

Further, the supplement is in the form of granule, tablet, powder, capsule, paste, microcapsule, or liquid.

The invention also provides a preparation method of the supplement, which comprises the following steps: mixing the pretreated lactoferrin, the beta-glucan and the auxiliary materials, and packaging into a product.

The invention also provides the application fields of the composition and the supplement, including but not limited to the application in the manufacture of medicaments, feeds and feed additives for preventing or treating animal herpesvirus.

The animal herpesvirus includes, but is not limited to, feline herpesvirus, bovine herpesvirus, equine herpesvirus, and avian herpesvirus.

The invention achieves the following beneficial technical effects: the composition and the animal food supplement provided by the invention take lactoferrin and beta-glucan as main functional components, and can play a good role in preventing and treating animal herpes, such as feline herpes virus, bovine herpes virus, equine herpes virus, poultry herpes virus and the like; the applicant finds in research that the surface of lactoferrin has positive charges, the surface of beta-glucan has negative charges, and the lactoferrin and the beta-glucan can be combined together through electrostatic adsorption, so that the lactoferrin can be loaded on the beta-glucan, particularly, the lactoferrin can be well prevented from being hydrolyzed by protease in stomach, successfully enters intestinal tracts and is combined with lactoferrin receptors on intestinal mucosa, and cells can pass through membranes to enter a circulatory system through receptor-mediated endocytosis to directly reach focuses; the beta-glucan has the function of improving immunity, and plays a role in adjuvant therapy by improving the immunity of the organism in the treatment process. The invention also provides the application of the lactoferrin and beta-glucan compositions and supplements in the preparation of medicaments, feeds and feed additives for preventing or treating animal herpesvirus, and through in vitro cell tests and animal tests, the compositions and supplements provided by the invention have good preventing and treating effects on animal herpesvirus such as feline herpesvirus (FHV-1); and the infection probability of healthy cats can be greatly reduced.

Detailed Description

The invention is further described with reference to specific examples. The following examples are only for illustrating the technical solutions of the present invention more clearly, and the protection scope of the present invention is not limited thereby.

The following examples are provided to further illustrate the present invention.

Example 1 preparation of animal food supplement

An animal food supplement, wherein the lactoferrin content is 0.1mg/g, 5mg/g, 20mg/g, 50mg/g, 60mg/g, 80mg/g, 100mg/g, 150mg/g, 180mg/g and 200mg/g, respectively, the percentage content of β -glucans is 10-800mg/g, preferably 250mg/g, and 500g supplements numbered 1, 2, 10, respectively, are obtained after addition of pre-treated auxiliary materials. The auxiliary materials are not essential to the invention, as long as the requirements of dosage form, taste and the like can be met, and therefore, the details are not repeated herein.

To illustrate the binding results between lactoferrin species and β -glucans of the present application, samples of the 1-10 supplement were analyzed for ￡ potential using a Zetasizer Nano-ZS90 nanometer laser particle sizer below. The measurement temperature was 25 ℃ and the measurement was carried out 3 times for each sample, and the results are shown in Table 1 as an average value.

TABLE 1 shows the results of ￡ potential testing for samples nos. 1-10

As can be seen from Table 1, the ￡ potential of lactoferrin species is + 4.77. + -. 0.5 mV; the surface of the beta-glucan has certain negative charges, as shown in the table 1, the-potential is ￡ 1.12 +/-0.01 mV, so that the lactoferrin and the beta-glucan can be combined together through electrostatic interaction. Moreover, as can be seen from table 1, as the content of the lactoferrin increases, the ￡ potential of the supplement changes and gradually rises, which indicates that electrostatic interaction occurs between the lactoferrin and the β -glucan to bond the lactoferrin and the β -glucan, that is, the lactoferrin is grafted to the β -glucan by the electrostatic interaction between the positive charge carried by the lactoferrin and the negative charge on the surface of the β -glucan. When the amount of lactoferrin is increased to 10%, the ￡ potential of the complex changes less, indicating that the binding site provided by the surface of the beta-glucan saccharide is completely covered by lactoferrin and that the lactoferrin becomes saturated.

Example 2 in vitro assay

1. Preparation of supplement: weighing different amounts of No. 7 supplements in 10ml of Phosphate Buffered Saline (PBS), and respectively preparing supplement solutions with the concentrations of 5mg/ml, 10mg/ml, 18mg/ml, 20 mg/ml and 25mg/ml and the labels of I, II, III, IV and IV; preparing 15mg/ml of lactoferrin as a lactoferrin control group, and preparing 15mg/ml of beta-glucan as a beta-glucan control group.

2. Reagents and materials: the herpesvirus is represented by cat rhinotracheitis virus (FHV-1), wherein the cat rhinotracheitis virus (FHV-1) and the cat kidney cell (F81) are both provided by laboratories of animal epidemic disease prevention and control centers in Nanjing; sodium bicarbonate, sodium chloride, sodium hydroxide, potassium chloride, disodium hydrogen phosphate, potassium dihydrogen phosphate, and DMSO are all analytical grade, purchased from Sigma company; fetal bovine serum, trypsin, DMEM medium were purchased from Gibco; MTT was purchased from Bio-sharp.

FHV-1 virulence test: collecting adherent F81 cells, digesting with pancreatin, suspending with cell culture solution, inoculating to 96-well plate with cell concentration of 2 × 10 and cell concentration of 100 μ L per well⁶And (4) per mL, and after the cells are completely attached to the wall in the incubator, discarding the supernatant. Adding 100 μ L of 10 times diluted virus solution into each well, setting each dilution as 8 wells, mixing, culturing at 37 deg.C in CO2 incubator for 72 hr, discarding the culture solution, observing cytopathic condition with microscope, and calculating TCID of half tissue cell infection amount of virus₅₀The results are shown in Table 2.

TABLE 2 measurement of TCID50 for FHV-1

Degree of dilution	Number of cytopathic wells/cell	Number of acellular lesion wells/cell	Rate of cellular disease
				10^-1	8	0	100％
10^-2	8	0	100％
				10^-3	8	0	100％
10^-4	5	3	62.50％
				10^-5	2	6	25％
10^-6	0	8	0％
				10^-7	0	8	0
10^-8	0	8	0

As can be seen from Table 2, FHV-1 showed significant cytopathic effects during the proliferation and virulence determination, the number of FHV-1 cytopathic wells inoculated at each viral dilution was counted, and the TCID50 of the virus was calculated to be 10 according to the Reed-Muench method^-4.3/0.1mL。

4. Cytotoxicity test: culturing F81 cells in a 96-well plate, removing supernatant after adherence, adding 100 mu L of supplement solution with the concentrations of I, II, III, IV and V, setting a control group, adding 10 mu L of prepared MTT after culturing for 24h, continuing culturing for 4h, removing supernatant, adding 100 mu L of DMSO, incubating for 30min at 37 ℃ in a CO2 incubator for dissolution, determining the D value of each well at 490nm by using an enzyme-labeling instrument after complete dissolution, repeating 3 times for each concentration of lactoferrin supplement solution, and taking the average value of the results. The cell survival rate (%) ═ D value of experimental group/D value of blank control group × 100%, the results are shown in table 3.

TABLE 3 toxic Effect of lactoferrin supplements on F81 cells

As can be seen from Table 3, the supplements, lactoferrin and β -glucans with different concentrations were directly reacted with F81 cells respectively, and then the cell survival rates were all 100% by formula calculation, indicating that the supplement of the present application is non-cytotoxic, and can be used in various fields such as food, and the like, for the prevention and treatment of FHV-1 virus.

5. And (3) virus prevention test: culturing F81 cells in 96-well plate until the cells adhere to the wall, discarding the supernatant, adding lactoferrin solution, beta-glucan solution, I, II, III, IV into each wellV.sub.50. mu.L of the supplement solution, incubation was continued for 6h, and then 50. mu.L of 100TCID was added to each well₅₀FHV-1 liquid is continuously cultured for 24h, 10 mu of LMTT is added, the culture is continuously carried out for 4h, 100 mu of LDMSO is added after the supernatant is discarded, and the mixture is incubated in an incubator for 30 min. D value at 490nm is measured by a microplate reader, and a normal cell control group and a virus control group are set, each concentration of lactoferrin supplement solution is repeated for 3 times, and the results are averaged to calculate cell survival rate (%).

TABLE 4 preventive effect of lactoferrin supplement on FHV-1 infection in F81 cells

As shown in table 4, the cell survival rate was greatly improved after 6h of exposure to F81 cells with the supplement compared to the non-supplemented virus group. As can be seen from the lactoferrin and beta-glucan control groups, the lactoferrin and beta-glucan are combined together, so that the effect of preventing FHV-1 virus is more obvious. Therefore, the applicant speculates that the supplement can effectively block the adsorption of virus to F81 cells in the early stage of FHV-1 virus infection, and further plays a role in preventing infection.

The reason for this is probably that the virus is adsorbed onto the host cell by recognizing the acetyl sulfate on the host cell by the gC glycoprotein specificity on the envelope of the virus during the process of adsorbing the virus onto the host cell, and inducing the envelope of the virus to bind with the host cell membrane specificity by the gD glycoprotein, so that the virus is firmly adsorbed onto the host cell. The combination of lactoferrin and heparan sulfate entering the organism can effectively organize the adsorption of viruses on host cells, thereby playing a role in preventing virus infection.

6. And (3) antiviral test: culturing F81 cells in 96-well plate for 24h, removing supernatant after cell monolayer adherence is complete, and adding 50 μ L of 100TCID into each well₅₀FHV-1 solution, and continuing to culture for 6 h. Adding lactoferrin solution, beta-glucan solution, and supplement solution (50 μ L) of I, II, III, IV and V into each well, placing in a CO2 incubator at 37 deg.C, and culturingAfter 48h, 10 mu of LMTT is added, the culture is continued for 4h, the supernatant is discarded, 100 mu of LDMSO is added, and the culture box is incubated for 30 min. The microplate reader measures the D value at 490 nm. The results were averaged in 3 replicates per concentration of lactoferrin supplement solution. And setting a normal cell control group and a virus control group, and calculating the virus inhibition rate according to a formula. The virus inhibition ratio (%) ((lactoferrin supplement group D value-virus control group D value)/(cell control group D value-virus control group D value) × 100%) was as shown in fig. 5.

TABLE 5 inhibition of FHV-1 by lactoferrin supplementation

From the results in Table 5, it can be seen that FHV-1 infected F81 cells treated with different concentrations of supplements, lactoferrin, and β -glucans showed an increase in cell survival with increasing concentration. The supplement provided by the application can effectively interfere replication and expression of virus in cells to cause cell rupture after F81 cells are infected by FHV-1, and the survival rate of F81 cells infected by the virus is greatly improved; meanwhile, as can be seen from the lactoferrin control group, the lactoferrin can interfere the replication expression of the virus in the virus replication phase, so that the lactoferrin has an antiviral effect, and the result is consistent with that of the supplement; the result of the beta-glucan control group shows that the beta-glucan has no obvious effect on inhibiting virus replication, and possibly the antiviral mechanism is to achieve the antiviral purpose by mobilizing the immunity of the organism. In addition, the research shows that after the virus infects host cells, the pathogenic mechanism is as follows: after the viral genetic genes enter the host cell, they are replicated in the host cell and then assembled, eventually resulting in the disruption of the host cell. As glycoprotein gB, which is essential in the replication of viruses, it is encoded by 948 amino acids and contains 10 glycosylation sites at the nitrogen terminus. Thus, if it interferes with the glycosylation modification of the gB glycoprotein, for example, prevents the binding of the sugar modifying the gB glycoprotein to the glycosylation site, thereby altering the final expression of the gB glycoprotein. While lactoferrin can act to inhibit viral replication, it is speculated that it may be because the glycosylation site contained on its surface competitively binds to the sugar required for the glycosylation of gB glycoprotein, causing the glycosylation modification process of gB glycoprotein to cease, thereby ending the viral replication process.

In conclusion, in vitro cell tests show that the supplement and lactoferrin provided by the invention can play a role in preventing and treating FHV-1. To further illustrate the technical effect of the present invention in the prevention and treatment of FHV-1, the following animal model is specifically described.

Example 3 animal experiments

Selecting 70 cats with nasus, wherein the variety of the cats is not limited and is divided into A group, B group, C group, D group, E group, F group and G group. 5 cats in each group were severely ill and showed symptoms of mental depression, anorexia, fever, blepharospasm, sneezing, lacrimation, significant purulent discharge of eyes and nose, and the remaining 5 cats showed only mild-moderate nasal discharge and/or lacrimation and/or sneezing; each group was added 5 healthy cats as controls.

TABLE 6 clinical sign grading Table of Cat infected with herpes Virus

The evaluation method comprises the following steps: during treatment, in order to avoid subjective errors of different observer personnel, all cats were evaluated by 1 observer every day, a semi-quantitative scoring system was used to grade and score clinical signs of infected cats with herpes virus according to the clinical sign grading table of the infected cats as shown in table 6, and the grading scores were updated in time according to the cat's disease symptoms.

The specific process is as follows: 1. summarizing the clinical sign grading scores of each cat in the monitoring period (every 7 days is one monitoring period) to obtain the total disease score of each cat, and taking the highest total disease score of each cat in the monitoring period; 2. the highest total score of all cats in each group was ranked in order, the median was taken as the median total score of disease, i.e., the median of the highest score of disease in each group was obtained, the difference between the median and the baseline was divided by the baseline value multiplied by 100% and the resulting value was taken as the degree of improvement (percentage) in the signs.

Application example 1: the lactoferrin supplement of example 2 was incorporated into the diet of a group A cat twice daily, weighing ≦ 2kg, 50-100mg each time; the weight is more than or equal to 2kg, and each time is 100-200 mg; the treatment and recurrence rates for cats lasting for one, two, three, four, and five weeks are shown in Table 7.

Application example 2: the lactoferrin supplement of example 2, No. ii, was incorporated into the diet of group B cats twice daily, weighing ≤ 2kg, 50-100mg each time; the weight is more than or equal to 2kg, and each time is 100-200 mg; the treatment and recurrence rates for cats lasting for one, two, three, four, and five weeks are shown in Table 7.

Application example 3: the lactoferrin supplement of example 2, No. iii, was incorporated into the diet of group C cats twice daily, weighing no more than 2kg, 50-100mg each time; the weight is more than or equal to 2kg, and each time is 100-200 mg; the treatment and recurrence rates for cats lasting for one, two, three, four, and five weeks are shown in Table 7.

Application example 4: the lactoferrin supplement of example 2, number IV, was incorporated into the diet of group D cats twice daily, weighing ≦ 2kg, 50-100mg each time; the weight is more than or equal to 2kg, and each time is 100-200 mg; the treatment and recurrence rates for cats lasting for one, two, three, four, and five weeks are shown in Table 7.

Application example 5: the lactoferrin supplement of example 2 was incorporated into the diet of group E cats twice daily, weighing ≤ 2kg, 50-100mg each time; the weight is more than or equal to 2kg, and each time is 100-200 mg; the treatment and recurrence rates for cats lasting for one, two, three, four, and five weeks are shown in Table 7.

Application example 6: the beta-glucans are added into the daily diet of the F group of cats twice a day, the weight is less than or equal to 2kg, and each time is 50-100 mg; the weight is more than or equal to 2kg, and each time is 100-200 mg; the treatment and recurrence rates for cats lasting for one, two, three, four, and five weeks are shown in Table 7.

Application example 7: adding lactoferrin into the diet of cat in group G twice a day, with a weight of 2kg or less and 50-100mg each time; the weight is more than or equal to 2kg, and each time is 100-200 mg; the treatment and recurrence rates for cats lasting for one, two, three, four, and five weeks are shown in Table 7.

As can be seen from table 7:

1. for mild cats, there was evidence of signs of improvement after one week of consumption of supplements I, II, III, IV and V. Five weeks after group a received supplement No. i, all cats recovered. B. After eating group C supplements ii and iii for four weeks, all cats healed and no recurrence occurred for the following one week. D. Group E was completely cured after one week of consumption of supplements iv and v, respectively, and subsequently monitored for two weeks without recurrence. From the aspect of sign improvement, the general condition that the group E is better than the group D is better than the group C is better than the group B is better than the group A. In the F group, some cats in the early stage of the edible beta-glucan control group have physical sign improvement conditions, but the symptoms of other cats are aggravated, but from the third week, all cats get worse, and possibly cats with better constitution can have immunity improved to a certain extent after taking in the beta-glucan, the physical signs are improved under the auxiliary action of the beta-glucan, but the later-stage virus replication speed is greater than the self-defense degree and the repair speed, and the condition of the cats is aggravated because of no anti-virus medicine intervention; after the beginning of gavage of the lactoferrin supplement at week four, cats began to improve and all appeared to have improved signs by week five. The symptoms of part of cats in the group G eating the lactoferrin control group are improved, the diseases of other cats are aggravated, the symptoms are improved in the later period, but the improvement degree is less obvious, the recovery is very slow, after the gastric lavage of the lactoferrin supplement is started for four weeks, the physical signs are obviously improved, and the cats are nearly healed in the fifth week.

2. For severely symptomatic cats, signs improved after one week consumption of supplements I, II, III, IV and V. A. Group B to the end of the monitoring period of the fifth week, and still cats were not cured, but the recovery progress was steadily progressing. After eating group C III for five weeks, all cats were healed; D. group E was completely cured by taking supplements IV and V for four weeks, respectively. No recurrence by week five. The overall situation was better for group D than group E than group C than group B. And when the F group eats the beta-glucan, the illness condition of all cats is not controlled, the illness condition is aggravated, the disease condition is still not improved until the cycle of the second week is ended, the symptoms are improved after the third week, the lactoferrin supplement is continuously infused in the stomach in the fourth and fifth weeks, and the cats are nearly healed in the fifth week. After the group G takes lactoferrin for one week, 40% of cats have improved signs, the rest cats get worse, 80% of cats have improved signs in the second week but have slow recovery degree, the rest cats still get worse, the third week begins to irrigate the stomach to supplement the lactoferrin supplement, all cats have improved signs, and the fourth and fifth weeks continue to irrigate the stomach until the fifth week is close to recovery.

3. For healthy cats, the beta-glucan control group of group F had 25% of cats had developed disease after week two, and after lactoferrin supplementation of the invention cats started in week three, all cats had developed disease and had no recurrence from week four to week five.

TABLE 7 treatment statistics for groups A-G Cats

In conclusion, the supplement of the present invention can effectively prevent feline rhinovirus infection; for the sick cat, the traditional Chinese medicine composition can play a good treatment role.

The present invention has been disclosed in terms of the preferred embodiment, but is not intended to be limited to the embodiment, and all technical solutions obtained by substituting or converting equivalents thereof fall within the scope of the present invention.

Claims

1. A composition characterized by: comprises lactoferrin and beta-glucan, wherein the lactoferrin comprises one or more of lactoferrin, lactoferricin derivatives and modified products of lactoferricin hydrolysate; the beta-glucan is one or more of beta-glucan and beta-glucan derivatives.

2. The composition of claim 1, wherein: the modified products of the lactoferricin, the lactoferricin derivative and the lactoferricin hydrolysate comprise at least one tryptophan;

and/or

The content ratio of (1, 3) glycosidic bond to (1, 4) glycosidic bond in the beta-glucan, beta-glucan derivative is 1: (1 to 4.2), wherein the content ratio of (1, 3) glycosidic bond to (1, 4) glycosidic bond is preferably 1: (2.4 to 2.6), more preferably, the ratio of the (1, 3) glycosidic bond content to the (1, 4) glycosidic bond content is 1: 2.5;

and/or

3. The composition of claim 1, wherein: the content of lactoferrin is 0.1-200mg/g, and the content of beta-glucan is 10-800 mg/g; preferably, the content of lactoferrin is 5-180mg/g, and the content of beta-glucan is 10-600 mg/g; preferably, the content of lactoferrin is 20-150mg/g, and the content of beta-glucan is 50-550 mg/g; more preferably, the content of lactoferrin is 100mg/g, and the content of β -glucan is 250 mg/g.

4. A food supplement for animals characterized by: the lactoferrin type-B-glucan-containing emulsion comprises lactoferrin, beta-glucan and auxiliary materials, wherein the lactoferrin comprises one or more of lactoferrin, lactoferricin derivatives and modified products of lactoferricin hydrolysate; the beta-glucan is one or more of beta-glucan and beta-glucan derivatives.

5. The supplement according to claim 4, characterized in that: the modified products of the lactoferricin, the lactoferricin derivative and the lactoferricin hydrolysate comprise at least one tryptophan;

and/or

6. The supplement according to claim 4, characterized in that: the content of lactoferrin is 0.1-200mg/g, and the content of beta-glucan is 10-800 mg/g; preferably, the content of lactoferrin is 5-180mg/g, and the content of beta-glucan is 10-600 mg/g; preferably, the content of lactoferrin is 20-150mg/g, and the content of beta-glucan is 50-550 mg/g; more preferably, the content of lactoferrin is 100mg/g, and the content of β -glucan is 250 mg/g.

7. The supplement according to any one of claims 4-6, characterized in that: the supplement is in the form of granule, tablet, powder, capsule, paste, microcapsule, or liquid.

8. A method of preparing a supplement according to any one of claims 4-6, characterized in that: mixing the pretreated lactoferrin, the beta-glucan and the auxiliary materials, and packaging into a product.

9. Use of a composition as claimed in any one of claims 1 to 3, a supplement as claimed in any one of claims 4 to 6 in the manufacture of a medicament, feed additive for the prophylaxis or treatment of herpesvirus infections in animals.

10. Use according to claim 9, characterized in that: the animal herpesvirus includes feline herpesvirus, bovine herpesvirus, equine herpesvirus, and avian herpesvirus.