CN110354139A - A kind of composition of porcine epidemic diarrhea resisting virus infection and its application - Google Patents
A kind of composition of porcine epidemic diarrhea resisting virus infection and its application Download PDFInfo
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
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Abstract
The invention discloses a kind of composition of porcine epidemic diarrhea resisting virus infection and its applications, the composition includes zinc oxide and glyceryl monolaurate, the mass ratio of the two is 0.5 ~ 2:1, have the effect of significantly inhibiting Porcine epidemic diarrhea virus duplication, reduce Porcine epidemic diarrhea virus virulence, it can be used as feed addictive or oral agents, overcome the disadvantages of immuning failure that vaccine inoculation is likely to occur and antiviral drugs are using caused virus variation, it is without any side effects to animal body, it can use safely.
Description
Technical field
The invention belongs to virus diseases of pigs Prevention Technique field, it is more particularly to a kind of porcine epidemic diarrhea resisting virus infection
Composition and its application.
Background technique
Currently, control and prevention of disease situation in China's is increasingly severe, and counter-measure is extremely limited, causes pig production to face huge
Challenge.The pig epidemic diarrhea (PED) as caused by Porcine epidemic diarrhea virus (PEDV) is that a kind of acute intestinal of pig infects
Disease, the recurrent obtburst within the scope of countries in the world.PEDV infection of the China in outburst the whole country in 2010, gives pig breeding industry band
Huge economic loss is carried out.The main clinic symptoms of PEDV infection are greatly lowered for feed intake, and diarrhea is in water sample and companion
It with vomiting, is even dehydrated when serious, the pig of each age and kind is likely to infection morbidity, the most serious by its harm
Be nursery-age pig, disease incidence is up to 100%, is currently to lead to one of main epidemic disease of piglet Deaths.Natural conditions
Under, the incubation period of Infection in Piglets PEDV is 1~2 day, incubation period of artificial challenge according to attack toxic dose difference 8~24 hours it
Between, the piglet body temperature in incubation period can increase, and then occur vomitting or the symptom of diarrhea.
So far, in addition to the inactivation of PEDV and attenuated vaccine, it there is no effectively preventing measure.Although most of pig farmer
It can prevent PED by vaccine inoculation, but due to vaccine quality is irregular etc., the effect after inoculation is also undesirable.
Under the highdensity breeding situations in China, large dosage of antiviral drugs abuse, leading to the speed of mutation of Strain has been more than epidemic disease
The speed of seedling development, extensive epidemic disease are broken out often.By giving the pig feeding functional battalion that safely and effectively anti-PEDV infects
Feeding substance is very necessary, and application prospect is good.
Glyceryl monolaurate is also known as lauric acid/dodecanoic acid monoglyceride, belongs to medium chain fatty acid ester, is naturally present in breast milk, coconut palm
In seed oil and palmetto, chemically synthesized product appearance is generally flakey or oily, white or lurid acinous knot
Crystalline substance is a kind of excellent food emulsifying agent, belongs to foodstuffs without toxicity additive.Glyceryl monolaurate has excellent emulsification function
With extensive antibacterial action.Zinc oxide is widely used on feed as a kind of effective preventing piglet diarrhea substance, but it is added
The excessive bring breeding pollution of dosage causes the concern of the public, and government uses rank as in feed addictive to zinc oxide
Section and dosage are made that strict regulations.Zinc oxide is cheap, convenient using holding, additive effect is obvious, and piglet can be improved
Anti-oxidation stress ability, Performance and immune function reduce the quantity of Escherichia coli in enteron aisle chyme.Although there is laurel at present
Acid glyceride has the report of inhibitory effect to togavirus, but the research in relation to the anti-PEDV of glyceryl laurate ester not yet appears in the newspapers
Road, the more research not about glyceryl monolaurate and the anti-PEDV infection of zinc oxide composites are reported.About compound single laurel
The patent of invention (CN201511015521.5, CN201510268143.5) of acid glyceride and zinc oxide is for zinc oxide
Preparation, is not directed to the application of porcine epidemic diarrhea resisting virus infection.And the patent in relation to glyceryl monolaurate is more collection
In feed it is mould proof, animal in terms of, the prior art does not disclose glyceryl monolaurate and zinc oxide group
Epidemic diarrhea virus can be fought by closing use.
Summary of the invention
The object of the present invention is to provide a kind of compositions of porcine epidemic diarrhea resisting virus infection, with high security,
Using convenient, high to Porcine epidemic diarrhea virus infection protective rate effect.
To achieve the goals above, technical measures adopted by the present invention are as follows:
A kind of composition of porcine epidemic diarrhea resisting virus infection, including zinc oxide and glyceryl monolaurate, zinc oxide
It is 0.5~2:1 with glyceryl monolaurate mass ratio.
Application of the above-mentioned composition in prevention and treatment Porcine epidemic diarrhea virus infection:
The zinc oxide of aforementioned proportion and glyceryl monolaurate composition are added directly to feed, additive amount is pannage
The 0.1%~0.3% of total weight；
Or pulvis is made in the zinc oxide of aforementioned proportion and glyceryl monolaurate composition and carrier, with 40~120mg/
The dosage of kg weight is administered orally to pig；
The carrier include montmorillonite, vermiculite, zeolite powder, white carbon black, defatted rice bran, maize cob meal, starch, in wheat bran
Any one or combinations thereof object；
Or the zinc oxide of aforementioned proportion and glyceryl monolaurate composition and solvent and auxiliary material are mixed and made into oral solution,
It is administered orally with the dosage of 40~120mg/kg weight to pig；
The solvent includes formic acid, acetic acid, dehydrated alcohol, propylene glycol, dimethyl sulfoxide, any one in Tween 80
Or combinations thereof object, auxiliary material include lactose, mannitol, arbitrary one kind or combinations thereof object in soluble starch.
Compared with prior art, the present invention has the following advantages and beneficial effects:
It is sweet by compounding mono laurate although 1, glyceryl monolaurate and zinc oxide have certain anti-PEDV ability
Grease and zinc oxide form composition, can give full play to the anti-PEDV effect of collaboration between the two, the zinc of one side zinc oxide from
Son is from viral internal in conjunction with amino acid, and glyceryl monolaurate is then in conjunction with peplos, and collaboration inhibits virus multiplication；Separately
On the one hand, zinc oxide and glyceryl monolaurate may combine production resin acid zinc in enteron aisle, protect enteric epithelium mucous layer.
2, the composition of porcine epidemic diarrhea resisting virus infection of the invention is with nutriment zinc oxide and mono laurate
It is formed based on glyceride by 0.5~2:1 building preparation, there is the significant effect for inhibiting PEDV duplication, reducing PEDV virulence
Fruit only need to add in feed or can play the role of by oral administration prevention and treatment piglet prevalence diarrhea virus infection, overcome
The disadvantages of immuning failure and antiviral drugs that vaccine inoculation is likely to occur are using caused virus variation, to animal body without appointing
What toxic side effect, can use safely.
Specific embodiment
Embodiment 1
Zinc oxide and glyceryl monolaurate composition make the inhibition of Porcine epidemic diarrhea virus (PEDV) in-vitro multiplication
With
(1) test material: African green monkey kidney cell (Vero cell) is used for the culture of Porcine epidemic diarrhea virus, PED disease
Malicious (YN plants) are separated by National agricultural microorganism key lab, Hua Zhong Agriculture University, are saved and presented；DMEM culture medium,
EDTA, pancreatin, fetal calf serum and dual anti-(mycillin) are purchased from GIBCO company, and zinc oxide and glyceryl monolaurate are purchased from
Sigma company.
(2) test test method: is divided into three test groups, a blank group, a zinc oxide group and a single laurel
Composition (zinc oxide and the mono laurate in composition of various dose is added in acid glyceride group, test group in the medium respectively
The weight ratio of glyceride is 1:1), wherein composition low dose group (adding according to 100 μm of ol/L culture mediums), composition middle dosage
Group (being added according to 1mmol/L culture medium), composition group (being added according to 10mmol/L culture medium), blank control group (culture medium
In do not add composition), zinc oxide group (by the addition of 50 μm of ol/L culture mediums), glyceryl monolaurate group is (by 50 μm of ol/L trainings
Support base addition).
The inhibition of PEDV in-vitro multiplication is tested: PED virus liquid being done 10 times with DMEM culture solution first and is serially diluted, is taken
10-2、10-3、10-4、10-5、10-6、10-7Six dilutions are inoculated in 96 orifice plates of cultured Vero cell monolayer respectively,
100 holes μ L/, each group of DMEM culture solution being separately added into containing various concentration composition set 37 DEG C containing 5%CO2In incubator
Continue to cultivate 48h, observes cytopathy.Lesion positive hole count in the cell hole of each dilution is counted, Reed-Muench is utilized
Method calculates each group of TCID50, more different groups of TCID50Value determine composition to the inhibiting effect of PEDV in-vitro multiplication,
Each group is done three repetitions.
(3) experimental result:
A, inhibiting effect of the composition to PEDV in-vitro multiplication
TCID of 1 composition of table to different virus50It influences
Table 1 is as the result is shown: the composition that various concentration is added in cell culture fluid significantly affects PEDV in Vero cell
On TCID50Value, with the increase of composition additive amount in cell culture fluid, TCID of the PEDV on Vero cell50Significantly
Decline illustrates that the proliferation in Vero cell that composition of the invention cultivates PEDV in vitro significantly inhibits.
In addition, the inhibiting effect that composition is proliferated PEDV is clearly more powerful compared with individually addition zinc oxide or glyceryl monolaurate,
Show that the anti-PEDV function and effect of collaboration of the two are more preferable.
B, influence of the composition to external PEDV vero cells infection vigor
2 composition of table is to PEDV vero cells infection OD450Influence
As shown in Table 2, the composition for various concentration being added in cell culture fluid significantly affects PEDV vero cells infection
OD450Value, with the increase of composition additive amount in cell culture fluid, the OD of Vero cell450Value dramatically increases, and illustrates this hair
Bright composition promotes the proliferation of Vero cell, enhances the ability of the anti-PEDV infection of Vero cell.In addition, with independent addition
Zinc oxide or glyceryl monolaurate are compared, and composition promotes Vero cell, the protective effect to the PEDV Vero cell infected
It becomes apparent from.
Embodiment 2
The protective effect of zinc oxide and glyceryl monolaurate composition to PEDV infection piglet
(1) materials and methods: 210 21 ages in days, the consistent weanling pig of weight and gender disposably weaned are divided at random
It is 7 groups: control group, PEDV group, PEDV+0.033% zinc oxide group, PEDV+0.067% glyceryl monolaurate group, PEDV+
0.1% composition group (weight ratio of zinc oxide and glyceryl monolaurate is 0.5:1), (oxidation of PEDV+0.2% composition group
The weight ratio of zinc and glyceryl monolaurate is 2:1), PEDV+0.3% composition group (zinc oxide and glyceryl monolaurate
Weight ratio is 2:1).3 repetitions (column) of every component, 10, every column.The basal diet of each group feeding is consistent.PEDV+0.03% oxygen
Change zinc group and PEDV+0.07% glyceryl monolaurate group piglet is fed respectively containing 0.033% zinc oxide and 0.067% single laurel
The basal diet of acid glyceride.PEDV+0.1% composition group, PEDV+0.2% composition group and PEDV+0.3% composition group
Piglet feeds the basal diet containing 0.1% composition, 0.2% composition and 0.3% composition respectively.And control group and PEDV group
The basal diet of 0.04% colistin sulfate pre-mixing agent of feeding addition.After feeding 7 days, to PEDV group, PEDV+0.033% oxygen
Change zinc group, PEDV+0.067% glyceryl monolaurate group, PEDV+0.1% composition group, PEDV+0.2% composition group and
PEDV+0.3% composition group piglet is with 104.5TCID50The PEDV (YN plants) of/mL attacks poison, every oral perfusion 5.0mL, then every
From raising, 3 days are observed continuously to off-test.The diarrhea situation of the piglet of observation test daily, tests slaughter experiment piglet of expiring
Enteron aisle lesion is observed, while counting dead pig number, calculates production performance and diarrhea rate.Composition leads to the inhibiting effect of PEDV
The expression quantity of PEDV structural gene N or M in real-time quantitative PCR detection intestine of young pigs mucous membrane are crossed to determine.The antiviral energy of piglet
Power, immune function etc. are expressed by antiviral gene and ELISA kit method measures.
(2) test result
A, growth performance
Influence of 1 composition of table to piglet growth performance
Note: colleague's shoulder mark difference lowercase letter indication difference is significant (P < 0.05).Following table is same.
As shown in Table 1, it is infected using composition of the present invention as the anti-PEDV of piglet, compared with PEDV group, feed
0.033% zinc oxide of middle addition, 0.067% glyceryl monolaurate, 0.1% composition, 0.2% composition, 0.3% composition
It is respectively increased piglet average daily gain 74%, 93%, 142%, 118%, 134% (P < 0.05), reduction feed-weight ratio 7.43%,
10.40%, 20.30%, 15.84%, 18.32% (P < 0.05), significantly reduce diarrhea rate 32.5%, 40.9%, 79.5%,
74.6%, 77.4% (P < 0.05).Therefore, using zinc oxide and glyceryl monolaurate composition, PEDV infection can be improved
The growth performance of piglet reduces the diarrhea rate of piglet, so that PEDV infection weaned piglets is restored to normal level, and effect is bright
It is aobvious to add zinc oxide or glyceryl monolaurate better than independent.
B, enteron aisle PEDV is proliferated
The influence that 2 composition of table expresses intestine of young pigs PEDV structural gene N and M
As shown in Table 2, added compared with PEDV group, in feed 0.033% zinc oxide, 0.067% glyceryl monolaurate,
0.1% composition, 0.2% composition, 0.3% composition respectively reduce the expression of Jejunum of Piglets PEDV structural gene N
33.20%, 50%, 82.52%, 83.11%, 84.45% (P < 0.05) reduces the expression of jejunum PEDV structural gene M
23.00%, the expression of colon PEDV structural gene N drops in 25.48%, 58.17%, 38.50%, 34.90% (P < 0.05)
Low 63.94%, 73.45%, 92.83%, 94.73%, 93.72% (P < 0.05), makes the table of colon PEDV structural gene M
Up to reducing 54.94%, 58.80%, 82.53%, 81.87%, 78.67% (P < 0.05).Therefore, it is individually added in feed
Although zinc oxide or glyceryl monolaurate can all reduce PEDV virulence, inhibit PEDV duplication, composition effect of the invention
More preferably.
C, the expression of enteron aisle antiviral gene and immune function
Influence of 3 composition of table to intestine of young pigs antiviral gene expression and immune function
As shown in Table 3, it is infected using composition of the present invention as the anti-PEDV of piglet, compared with the control group, feed
The composition of middle addition 0.2%, 0.4%, 0.6%, which can be improved, reduces containing for blood inflammatory factor IL-1 β, IL-8 and IL-6
Amount alleviates inflammatory reaction caused by PEDV infects.And zinc oxide or glyceryl monolaurate alleviation are individually added in feed
The inflammatory reaction effect of PEDV induction is not so good as composition of the invention.In addition, adding 0.1%, 0.2%, 0.3% group in feed
It closes object and also reduces the expression of enteron aisle ISG15, MX1 and S100A12, show that the present composition inhibits answering for enteron aisle PEDV
System, reduces the virulence of PEDV, is at a normal level intestine of young pigs antiviral protein, promote the intestinal health of piglet.And it is single
Solely addition zinc oxide reduced by only the expression of enteron aisle S100A12, to the expressional function effect of ISG15 and MX1 not as good as of the invention
Composition.It can be seen that compounding the anti-PEDV infection composition formed by zinc oxide and glyceryl monolaurate, can give full play to
The anti-PEDV infection effect of the collaboration of the two.
Embodiment 3
A kind of composition of porcine epidemic diarrhea resisting virus infection and its application, the steps include:
(1) glyceryl monolaurate 10g, montmorillonite 30g is weighed to be uniformly mixed；
(2) it weighs 20g zinc oxide to be added in step (1) mixture, be uniformly mixed；
(3) obtain a kind of porcine epidemic diarrhea resisting virus infection mixture (mass ratio=1 of composition and carrier:
1), wherein zinc oxide and glyceryl monolaurate are 2:1 by weight in composition；
(4) it is used as usually prevention Porcine epidemic diarrhea virus infection, by mixture according to feed total weight 0.2%
Additive amount is added into feed (additive capacity of practical glyceryl monolaurate and zinc oxide composition is 0.1%), entire
Breeding cycle uses；
It (5) for clinical Porcine epidemic diarrhea virus treatment of infection, can be by mixture according to feed total weight when
0.6% dosage is added into feed feeding, and (additive capacity of practical glyceryl monolaurate and zinc oxide composition is
0.3%) it, adds three times, is used continuously one week in the morning, afternoon and evening daily.
Embodiment 4
A kind of composition of porcine epidemic diarrhea resisting virus infection and its application, the steps include:
(1) glyceryl monolaurate 100g, zinc oxide 50g are weighed, is uniformly mixed；
(2) a kind of composition of porcine epidemic diarrhea resisting virus infection is obtained, wherein zinc oxide and glyceryl monolaurate
It is by weight 0.5:1；
(3) composition 12g, 5% acetic acid 5mL, dehydrated alcohol 15mL, Tween-80 30mL, water for injection is weighed to add to
100mL is uniformly mixed；
(4) a kind of composition oral liquid (120mg/mL) of porcine epidemic diarrhea resisting virus infection is obtained；
(5) as usually prevention Porcine epidemic diarrhea virus infection use, can according to 40mg/kg weight to pig take orally to
Medicine is used in entire breeding cycle；
It (6) for clinical Porcine epidemic diarrhea virus treatment of infection, can be by oral solution according to 120mg/kg body when
Relatively heavy amount is administered orally directly to pig；Or mix oral solution with daily ration according to 120mg/kg body weight dose, it is added into feed
It uses, adds three times, be used continuously one week in the morning, afternoon and evening daily.
Embodiment 5
A kind of composition of porcine epidemic diarrhea resisting virus infection and its application, the steps include:
(1) glyceryl monolaurate 100g, zinc oxide 100g are weighed, is uniformly mixed；
(2) a kind of composition of porcine epidemic diarrhea resisting virus infection is obtained, wherein zinc oxide and glyceryl monolaurate
It is by weight 1:1；
(3) composition 50g, soluble starch 50g or wheat bran 50g are weighed, is uniformly mixed, 60 meshes are crossed, is dispensed up to powder
Agent；
(4) it uses, can be administered orally according to the dosage of 40mg/kg weight to pig, whole as when usually preventing swine disease
A breeding cycle uses；
(5) when being used for clinical treatment swine disease, pulvis can be taken orally to pig according to the dosage of 120mg/kg weight
Administration；Or pulvis is soluble in water according to the dosage of 120mg/kg weight, or it is added into feed use, it adds in the morning, afternoon and evening daily
Three times, it is used continuously one week.
Claims (5)
1. a kind of composition of porcine epidemic diarrhea resisting virus infection, which is characterized in that including zinc oxide and glycerol monolaurate
Ester, the mass ratio of the two are 0.5 ~ 2:1.
2. the composition of porcine epidemic diarrhea resisting virus infection according to claim 1, which is characterized in that further include carrying
Body, the carrier include montmorillonite, vermiculite, zeolite powder, white carbon black, defatted rice bran, maize cob meal, starch, appointing in wheat bran
One kind or combinations thereof of anticipating object.
3. the composition of porcine epidemic diarrhea resisting virus infection according to claim 1, which is characterized in that the combination
Object is oral solution, further includes solvent.
4. composition described in any one of claims 1 to 3 is in the drug of preparation prevention and treatment Porcine epidemic diarrhea virus infection
Application.
5. application according to claim 4, which is characterized in that zinc oxide and glyceryl monolaurate composition directly add
Into feed, additive amount is the 0.1% ~ 0.3% of pannage total weight；Or powder is made in zinc oxide and glyceryl monolaurate composition
After agent or aqua, it is administered orally with the dosage of 40 ~ 120mg/kg weight to pig.
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| CN114521617A (en) * | 2022-03-21 | 2022-05-24 | 江苏农牧科技职业学院 | Feed additive for preventing pig diarrhea and preparation method thereof |
| CN114521617B (en) * | 2022-03-21 | 2023-11-07 | 江苏农牧科技职业学院 | Feed additive for preventing pig diarrhea and preparation method thereof |
| CN115645417A (en) * | 2022-10-17 | 2023-01-31 | 武汉轻工大学 | Application of chenodeoxycholic acid in preparation of medicine or feed additive for treating or preventing porcine viral diarrhea |
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