CN106562037A - Honeysuckle stem and leaf feed additive for preventing and treating pig intestinal tract infectious diseases and improving feed utilization ratio as well as preparation method and application thereof - Google Patents
Honeysuckle stem and leaf feed additive for preventing and treating pig intestinal tract infectious diseases and improving feed utilization ratio as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN106562037A CN106562037A CN201610821184.7A CN201610821184A CN106562037A CN 106562037 A CN106562037 A CN 106562037A CN 201610821184 A CN201610821184 A CN 201610821184A CN 106562037 A CN106562037 A CN 106562037A
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- Prior art keywords
- feed additive
- feed
- leaf
- stem
- flos lonicerae
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- LSQZJLSUYDQPKJ-UHFFFAOYSA-N p-Hydroxyampicillin Natural products O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)C(N)C1=CC=C(O)C=C1 LSQZJLSUYDQPKJ-UHFFFAOYSA-N 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 229940055726 pantothenic acid Drugs 0.000 description 1
- 235000019161 pantothenic acid Nutrition 0.000 description 1
- 239000011713 pantothenic acid Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 229960000329 ribavirin Drugs 0.000 description 1
- HZCAHMRRMINHDJ-DBRKOABJSA-N ribavirin Natural products O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1N=CN=C1 HZCAHMRRMINHDJ-DBRKOABJSA-N 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 230000008313 sensitization Effects 0.000 description 1
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical class [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-L sulfite Chemical compound [O-]S([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-L 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 125000003831 tetrazolyl group Chemical group 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 230000002257 verocytotoxic effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/35—Caprifoliaceae (Honeysuckle family)
- A61K36/355—Lonicera (honeysuckle)
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
- Fodder In General (AREA)
Abstract
The invention provides a feed additive for preventing and treating pig intestinal tract infectious diseases and improving feed utilization ratio. The feed additive is prepared from the raw materials consisting of honeysuckle stems and leaves, is mechanically-pulverized micro powder or extract and is capable of effectively preventing and treating pig intestinal tract infectious diseases, and especially preventing and treating porcine diarrhea. The feed additive is also capable of promoting the digestion function, improving the microflora balance of pig intestinal tracts and improving the feed absorption capacity and the feed utilization ratio. The honeysuckle stems and leaves selected in the raw materials for the feed additive are abundant in source, the honeysuckle stem and leaf feed additive is stable in quality and low in cost, and the preparation method is simple in preparation process, free from pollution and suitable for industrial production and has favorable economical and social benefits.
Description
Technical field
The invention belongs to livestock feed additive agent field, the present invention relates to one kind is in preventing and treating Intestinum Sus domestica road transmission disease and carries
The feed additive of high efficiency of feed utilization, feedstuff, preparation method and applications.
Background technology
In recent years, with the raising of living standards of the people, Carnis Sus domestica consumption figure also increases year by year, and then drives domestic aquaculture
Fast development.But pig disease frequently breaks out serious prestige evil pig industry, wherein diarrhea of pigs accounts for the 35- of pig disease total amount
45%, it is the high incidence disease that may occur throughout the year, while can also cause other viruses or antibacterial concurrent or secondary
Infection.Up to now, it has been found that in the various causes of disease of induction diarrhea of pigs disease, with the infectious disease that antibacterial and virus cause
Harm is the most serious, common are Porcine epidemic diarrhea virus, transmissible gastro-enteritis viruss, rotavirus illness, pathogenic big
Enterobacteria etc., mixed infection is there is also, and it is in rising trend, especially large-scale intensive pig farm harm is even more serious.Sick pig
While being mainly shown as diarrhoea, poor growth, the price of deed is reduced, and production performance declines, or even causes death.
At present for the preventing and treating of diarrhea of pigs is mainly glued using gentamycin, norfloxacin, enrofloxacin, amoxicillin, sulphuric acid
The antibiotic medicines such as rhzomorph, Ciprofloxacin and florfenicol.Due to life-time service or abuse of antibiotics, its negative effect is also increasingly
Appear, its result not only causes immunologic hypofunction, destroy gastrointestinol microorganism ecological balance, disturb it normally to digest and assimilate
Function, the also bioavailability of appreciable impact feedstuff, the problems such as cause antibacterial to produce drug resistance and antibiotic remainss.
People's long-term consumption has the pork product of antibiotic remainss, easily causes allergic reaction, toxic reaction etc., and harm is strong
Health.And the antiviral drugs such as amantadine, IFN-γ and oseltamivir phosphate capsule, because the reasons such as cost height are difficult to be received by raiser.Although
Vaccine tool has certain effect, but prevents single, and effect and unstable, it is difficult to the quick variation of correspondence virus.Therefore, it is a kind of
Can efficiently, noresidue, without drug resistance, antibacterial, antiviral and anti-infectious function prevention and treatment diarrhea of pigs medicine and composition
Thing is undoubtedly the urgent needss of aquaculture.
The content of the invention
To solve the deficiencies in the prior art, the present invention provide it is a kind of it is with antibacterial, antiviral and anti-infectious function, with gold
Flos Lonicerae stem and leaf is the preventing and treating Intestinum Sus domestica road transmission disease of raw material composition and the feed additive for improving efficiency of feed utilization.
The feed additive includes stem and leaf of Flos Lonicerae micropowder or extract, for preparing Intestinum Sus domestica road transmission disease, especially
It is the feed additive for preventing and treating porcine epizootic diarrhea, the feed additive can play one to the development of intensive pig aquaculture
Fixed facilitation.
Although the effect such as Flos Lonicerae and the antibacterial of active component, antiviral and infection is known, up to now
There is not yet being used for the preventive and therapeutic effect of diarrhea of pigs for the micropowder and extract of raw material using stem and leaf of Flos Lonicerae.And the feedstuff of the present invention
Additive raw material wide material sources, preparation process is simple, low production cost, using feasible, definite effect, will not produce drug resistance,
Porcine epizootic diarrhea disease can safely and effectively be prevented and treated.The feed additive of the present invention is used to prevent and treat the treatment of Intestinum Sus domestica road transmission disease
Imitate, drug residue free, have no drug resistance, have no toxic side effect.
The invention further relates to prepare the feedstuff addition of described preventing and treating Intestinum Sus domestica road transmission disease and raising efficiency of feed utilization
The method of agent.
The preparation method of the present invention is simple, with low cost, and during being pruned with plant is seasonal stem and leaf of Flos Lonicerae is obtained
Prepare.
The invention further relates to the addition of the preventing and treating Intestinum Sus domestica road transmission disease and improve the feedstuff addition of efficiency of feed utilization
The feedstuff of agent.
The invention further relates to the feed additive of the preventing and treating Intestinum Sus domestica road transmission disease and raising efficiency of feed utilization is in system
Application in the medicine of standby preventing and treating Intestinum Sus domestica road transmission disease.
The feed additive of the present invention can be used to prepare the medicine of the various Intestinum Sus domestica road transmission diseases of preventing and treating, particularly be used for
Prepare the medicine of preventing and treating porcine epizootic diarrhea.
The feed additive of the present invention can improve Gaster Sus domestica intestinal microbial population balance and safeguard environment in intestinal physiology, improves right
Application in terms of feedstuff absorbability and efficiency of feed utilization.
The feed additive of the present invention can safeguard environment in Gaster Sus domestica intestinal physiology, improve the adaptability to environment and support
Grow survival rate.
Description of the drawings
Fig. 1 is stem of Flos Lonicerae, leaf and the liquid phase analysis contrast of flower Content of Chlorogenic Acid；
Fig. 2 is the molecular weight of each main peak in leaf of Flos Lonicerae；
Fig. 3 is the active component contained in leaf of Flos Lonicerae.
Specific embodiment
Hereinafter, embodiments of the present invention are illustrated.
The present invention relates to the feed additive of a kind of preventing and treating Intestinum Sus domestica road transmission disease and raising efficiency of feed utilization, the feeding
Feed additives raw material is made up of the stem or/and leaf of Flos Lonicerae；Gold containing percentage by weight 50～100% in the feed additive
Flos Lonicerae stem and leaf micropowder or extract.
Result of study prompting stem and leaf of Flos Lonicerae mechanical activation comminution micropowder of the present invention or extract are added into after feedstuff, to pig disease
With significant prevention effect, pig antibacterial and disease of viral infection can be prevented and treated as feed additive.
Used as one embodiment of the present invention, the feed additive contains 100% stem and leaf of Flos Lonicerae micropowder, the gold silver
The particle diameter of scape leaf micropowder powder is less than 0.85mm；Or Flos Lonicerae of the feed additive containing percentage by weight 50～100%
Stem and leaf water extract, per gram of the stem and leaf of Flos Lonicerae water extract contains crude drug>3g.
The preventing and treating pig antibacterial and disease of viral infection feed additive that the present invention is provided is to prune Flos Lonicerae with season to plant
The stem and leaf obtained during thing is made through mechanical activation comminution as raw material into powder, and the particle diameter of powder is less than 0.85mm (20 mesh), with
It is easy to digesting and assimilating for domestic animal, and reduces the infringement to domestic animal digestive system.When in use, the feed additive of powder is mixed
It is incorporated in feedstuff and stirs, the feedstuff mildew-resistant additive is suitable than 1～5% with the weight of feedstuff.
Used as one embodiment of the present invention, micropowder containing stem and leaf of Flos Lonicerae or extract are 50 in the feed additive
～100%.
The present invention relates to a kind of described preventing and treating Intestinum Sus domestica road transmission disease and feedstuff for improving efficiency of feed utilization of preparing adds
Plus the method for agent, wherein, methods described includes：Powder by the raw material through mechanical activation comminution into particle diameter less than 0.85mm, obtains
To the feed additive.
The present invention relates to it is a kind of prepare the described transfecting disease in preventing and treating Intestinum Sus domestica road and improve the feedstuff of efficiency of feed utilization add
Plus the method for agent, wherein, methods described includes：Step (1), by the raw material pulverizing into powder；Step (2), by the step
(1) the material powder addition solvent for obtaining, the solvent is water, ethanol or aquiferous ethanol, and the solvent is water, ethanol or aqueous
Ethanol, the weight ratio of described material powder and solvent is 1:1～20, the solution of acquisition by heating extraction, microwave extraction or
Supersound extraction obtains the feed additive.
In view of stem of Flos Lonicerae leaf extract is developed as final products using feed additive, therefore consider from safety perspective
Extraction solvent selects water, ethanol or aquiferous ethanol to be advisable as far as possible.The ratio of stem and leaf of Flos Lonicerae and solvent does not have specific limit during extraction
System, but the ratio of general stem and leaf of Flos Lonicerae and solvent is 1:1～20 scope is advisable.The present invention does not have specific limit to Extracting temperature
System, it is all applicable to solvent boiling point scope from room temperature.Certainly room temperature, normal pressure and extract in the range of solvent boiling point ideal.
Extracting method can using heating extraction, supersound extraction, microwave extraction etc., also can various methods combine application.During extraction
Between from 30 minutes to 24 hours, scope can be with.
The present invention relates to a kind of described preventing and treating Intestinum Sus domestica road transmission disease and feedstuff for improving efficiency of feed utilization of preparing adds
Plus the method for agent, wherein, methods described includes：Step (1), by the raw material pulverizing into powder；Step (2), by the step
(1) the material powder addition water for obtaining, described material powder and the weight ratio of water is 1:1～20, heating extraction, extraction time
For 30 minutes to 24 hours, the extracting solution is filtered；Step (3), by the filtering residue addition water after the filtration in the step (2),
Described filtering residue is 1 with the weight ratio of water:1～20, heating is extracted for second, and extraction time is 30 minutes to 24 hours, is filtered
Second extracting solution；Step (4), merges the filtrate that the step (2) and step (3) are obtained, and is concentrated to give described feeding
Feed additives.
Used as one embodiment of the present invention, the present invention prepares described preventing and treating Intestinum Sus domestica road transmission disease and raises with improving
In the method for the feed additive of material utilization rate, heating-up temperature is boiling temperature in the step (2) and step (3), during heating
Between be 30 minutes.
The invention further relates to the feedstuff of a kind of preventing and treating Intestinum Sus domestica road transmission disease and raising efficiency of feed utilization, the feedstuff bag
Weight is included than the feed additive described in 1～5%.
According to the preparation method of described two extracts obtain stem of Flos Lonicerae, leaf extract be powder when, anti-corrosion is prevented
Mould feed additive is mixed in feedstuff and stirs, and the feed additive is 1～5% with the weight ratio of feedstuff；
When feed additive is liquid extract, after first feed additive being diluted to into 40 times of volume multiple, then it is spilled in feedstuff
And stir, the feed additive is still suitable for 1～5% with the weight ratio of feedstuff.
Used as one embodiment of the present invention, the feed additive by mixing addition or described with the feedstuff
Feed additive is by being coated to the feed surface addition.
The invention further relates to described feed additive is in the medicine for preparing the prevention and treatment Intestinum Sus domestica road transmission disease
Application in thing, the Intestinum Sus domestica road transmission disease include the popular diarrhoea of yellow and white dysentery of piglet, pig, Escherichia coli infection or
Rotavirus infection and the gastroenteritiss because of diarrhoea induction.
Used as one embodiment of the present invention, the infective pathogen of the Intestinum Sus domestica road transmission disease includes pig epidemic
Diarrhea virus, transmissible gastro-enteritis viruss, rotavirus illness, Escherichia coli, staphylococcus aureuses, Candida albicans
Bacterium, bacillus pyocyaneus, dysentery bacterium, enterobacteria.
The invention further relates to described feed additive is improving Gaster Sus domestica intestinal microbial population balance, improve to feedstuff absorbability
With the application in terms of efficiency of feed utilization.
The present invention prunes stem, leaf that plant obtains etc. for steady quality, the raw material for easily obtaining to prepare feeding using Flos Lonicerae
Feed additives, using stem of Flos Lonicerae, leaf rich in the antimicrobial antiviral activity composition such as chlorogenic acid, it is to avoid making for antibiotic medicine
With, there is low cost, drug residue free, have no drug resistance, have no toxic side effect, process is simple during preparation, consume energy it is low, without dirt
Dye, is very easy to realize industrialization, new purposes is provided for Flos Lonicerae producing region, with good economic and social benefit.
With reference to example is embodied as, the present invention will be further described.
The stem of Flos Lonicerae leaf extract Active Components of embodiment 1
Analytical tool and raw material：
Thermostatic drying chamber is Germany's Memmert types
Electronic balance Sartorius BP211D, a ten thousandth of precision ten
Pulverizer Hangzhou Xiong Huo Science and Technology Ltd.s
Ultra-pure water instrument Merck millipore companies
High performance liquid chromatograph Dionex Ultimate 3000 are serial for analytical type
Low Resolution Mass Spectra instrument Bruker amazon SL ion trap mass spectrometers
Analysis chromatographic column is the μ C18 110A (5 μm, 250 × 4.6mm) of Phenomenex Gemini 5
Stem and leaf of Flos Lonicerae sample is provided by Julu capital ancient cooking vessel Murad Flos Lonicerae industrial park.
The Active Components of leaf of Flos Lonicerae adopt HPLC-MS methods, and need testing solution is prepared as respectively extracting honeysuckle powder
End, leaf of Flos Lonicerae powder, gold silver bennet powder 0.5g, accurately weighed, in being placed in conical flask, precision adds water 50ml, weighed quality,
After immersion 30min, decocted by heating and refluxing extraction method and be allowed to the 15min that seethes with excitement, let cool, then weighed quality, add water and supply less loss
Quality, shake up, filter, precision measures subsequent filtrate 10ml, water bath method, and with 1ml water dissolution, 0.45 μm of Jing after centrifugation are distilled
Water film filtering is used for HPLC-MS Instrumental Analysis.Chlorogenic acid reference substance is 0.46mg, plus methanol is configured to 46.0 μ g/ml reference substances
Solution is for this experiment.Experiment Detection wavelength be 365nm, column temperature be 40 DEG C, sample size be 20 μ l, mobile phase：Acetonitrile-
0.05% glacial acetic acid, using linear gradient elution program：0-60min acetonitriles 1% -30%, 0.05% glacial acetic acid 99% -
70%, flow velocity is 0.9ml/min.
Experimental result is as shown in Figure 1.
It can be seen from figure 1 that as can be seen that in leaf of Flos Lonicerae in stem of Flos Lonicerae, the comparison of ingredients of the extract of Ye Hehua
Chemical composition content is significantly higher than the extract of flower and stem, and various active composition (referring to Fig. 2) is contained in leaf of Flos Lonicerae extract,
Wherein it is mainly caffeoylquinic acids (caffeoylquinic acid), chlorogenic acid (chlorogenic acid), sweroside
(sweroside) and luteoloside (galuteolin) etc. is antibacterial, antiviral and oxidation-resistant active ingredient (referring to Fig. 3),
Suitable for the anti-cause of disease purposes of feedstuff, and negative effect will not be formed to the health of domestic animal.
Additionally, according to HPLC liquid chromatograph area of pictural surface ratio, the chlorogenic acid and luteoloside content in Flos Lonicerae, stem, leaf
As shown in table 1.
The content of the stem and leaf of Flos Lonicerae Content of Chlorogenic Acid of table 1 and luteoloside
| Experiment group | Flos Lonicerae | Leaf of Flos Lonicerae | Stem of Flos Lonicerae |
| Chlorogenic acid (%) | 6.17±0.12 | 5.89±0.15 | 4.60±0.32 |
| Luteoloside (%) | 0.11±0.02 | 0.35±0.01 | 0.16±0.01 |
Therefore, stem and leaf of Flos Lonicerae has suitable medical value.
The preparation of the feed additive of the present invention of embodiment 2
The feed additive of the present invention can be two kinds of forms of micropowder and extract.
The preparation of 2.1 stem of Flos Lonicerae, leaf micropowder
The stem obtained when pruning honeysuckle plant using season or/and leaf are dried, through mechanical activation comminution as raw material, cleaning
Make into powder, the particle diameter of powder is less than 0.85mm (20 mesh), obtains stem and leaf of Flos Lonicerae micropowder of the present invention..
The preparation of 2.2 stem of Flos Lonicerae, leaf water extract
The stem obtained when pruning honeysuckle plant using season or/and leaf are prepared from according to the following steps as raw material：
Step (1), by raw material pulverizing into powder；
Step (2), the material powder that step (1) is obtained adds the water of 10 times of amounts, is heated to boiling, extracts 30 minutes,
Filter extracting solution；
Step (3), by the filtering residue after the filtration in step (2) water of 10 times of amounts is added, and is heated to boiling, is carried for the second time
Take, extract 30 minutes, filter second extracting solution；
The filtrate that step (4), combining step (2) and step (3) are obtained, contains crude drug 3.2g in every gram of concentrate, obtain
Stem of Flos Lonicerae of the present invention or/and leaf water extract..
The stem and leaf of Flos Lonicerae of the present invention can also use ethanol, aquiferous ethanol to be extracted as solvent, gold silver during extraction
The ratio of scape leaf and solvent is 1:1～20 scope is advisable；Extracting temperature is all applicable to solvent boiling point scope from room temperature.Extract
Method can using heating extraction, supersound extraction, microwave extraction etc., also can various methods combine application.Extraction time from
30 minutes to 24 hours scopes can be with.
Feed additive of the present invention typically with the forms such as micropowder, extractum and microgranule be used for prevent and treat poultry antibacterial and
Disease of viral infection, its product can add conventional excipient, antioxidant, preservative, coloring agent in preparation process
Deng additive.Wherein, excipient can select starch, cellulose etc.；Preferred antioxidant has sulphite, ascorbic acid etc.；
Dihydroxy acetic acid, methaform etc. can be used as preservative.Stem and leaf of Flos Lonicerae micropowder or extract in the feed additive
Content is suitable 50～100%.
The anti-microbial pathogen effect of the feed additive stem of Flos Lonicerae leaf extract of the present invention of embodiment 3
Activity determination adopts the scraps of paper-agar diffusion method sensitization test (Kirby-Bauer methods), and its principle is according to antibacterial ring size
Diameter carries out activity rating with the dependency of minimum inhibitory concentration.Experiment uses Mueller-Hinton agar, prepares thick
Spend the agar plate for 4mm and pH7.2～7.4.After agar solidification, 4 DEG C of preservations are positioned over, and experiment is provided and use.Before experiment,
Lyophilizing bacterial strain is brought back to life with 1.5 × l08/ m1 seed agar pipes, prepare each bacterial suspension during experiment, in being suspended in normal saline,
It is 10 that vibration mixes post-equalization suspension concentration7CFU/ml.During inoculation flat board, bacterium solution even spread is dipped with the cotton swab of sterilizing whole
Individual media surface.Laboratory sample carries a diameter of 6mm of the scraps of paper, and leaf of Flos Lonicerae extraction prepared by extraction embodiment 2 is distinguished per piece
Thing, the μ l of stem extract amount about 20, are attached to drug sensitive test paper on the flat board for coating bacterium solution with aseptic nipper, middle heart septum between the scraps of paper
30mm, plate is placed in after cultivating 24 hours in 37 DEG C of calorstats, detects antibacterial circle diameter, and every kind of sample sets 6 samples and is averaged
Value.Experimental result is referring to table 1A, table 2.
The minimal inhibitory concentration (MIC) of table 1A leaf of Flos Lonicerae extract
The minimal inhibitory concentration (MIC) of the stem of Flos Lonicerae extract of table 2
From table 1A, 2 experimental results, leaf of Flos Lonicerae extract, stem extract can effectively suppress staphylococcus aureuses,
Candida albicans, bacillus pyocyaneus, escherichia coli, dysentery bacterium, enterobacteria.
The antiviral effect in vitro of the stem of Flos Lonicerae leaf extract of the present invention of embodiment 4
Experiment uses DMEM (Dulbecco ' s modified eagle medium) cell culture fluid D- containing 4500mg/L
Glucose, 300mg/L L-glutaminate, 110mg/L Sodium Pyruvates, 5% hyclone, 1% penicillin, 1% streptomycin.Will
After experiment Vero cells are with 0.25% pancreatin digestion, piping and druming disperses cell, and with DMEM culture fluid 1 × 10 is diluted to6Cell/
Ml, in adding to 96 porocyte culture plates, puts 5%CO2, cultivate in 37 DEG C of incubators, after cell grows up to cell monolayer, carry out body
Outer antivirus test.Cell dyeing adopts tetrazolium-based colorimetric assay (mtt assay), microplate reader to determine the extinction at 490nm
Angle value (OD), calculates suppression ratio.
The leaf of Flos Lonicerae extract of 4.1 present invention, the Vero cytotoxic effects of stem extract：
It is degerming with 0.22 μm of filter membrane after stem of Flos Lonicerae leaf extract PBS dissolvings of the invention prepared by embodiment 2, use
After cell culture fluid continuous gradient dilutions, in being respectively added to Vero cells (purchased from U.S. ATCC) culture plate, each concentration is carried
Take thing and set 4 culture holes, put 5%CO2, cultivate 72 hours under the conditions of 37 DEG C, observation of cell form under inverted microscope uses MTT
Method detects cytotoxicity, and determines drug action concentration.
Cytotoxic assay result shows that stem, leaf extract have not in the activity of more than 50 μ g/ml to cellular morphology
With the impact of degree, and in below 50 μ g/ml without obvious cytotoxicity.
Inhibitory action of the leaf of Flos Lonicerae extract, stem extract of 4.2 present invention to PEDV cytopathogenic effects：
Test Porcine epidemic diarrhea virus (porcine epidemic diarrhea virus, PEDV-GDZ-2014)
(the tropical medicine research of Guangdong university of TCM is separated and give) is inoculated in Vero cell monolayers, puts 5%CO2, 37 DEG C of cultures
72 hours, Ultrasonic Pulverization after infection cell is harvested, -40 DEG C of preservations after virus liquid subpackage, PEDV strains titre is 5 × 107PFU/ml。
Vero cells are inoculated with after 10 times of gradient dilutions, are evaluated for external anti-PEDV virus activities.Comparison medicine ribavirin injection 100g/
L (Jiangsu Xinghua pharmaceutical factory).
4.2.1 leaf of Flos Lonicerae extract of the invention, stem extract are to viral direct effect：
The stem of Flos Lonicerae leaf extract of variable concentrations is mixed with PEDV viruses, after room temperature is acted on 4 hours, is respectively added to
In Vero Tissue Culture Plates, each concentration extract sets 4 culture holes, puts 5%CO2, after cultivating 30 minutes under the conditions of 37 DEG C, remove
Culture fluid is removed, is washed with PBS three times, and add cell culture fluid in 5%CO2, cultivate 72 hours under the conditions of 37 DEG C, use
The antiviral activity of mtt assay Detection and Extraction thing.
4.2.2 the inhibitory activity of of the invention leaf of Flos Lonicerae extract, stem extract to virus multiplication：
By in PEDV virus inoculations to Vero Tissue Culture Plates, each concentration extract sets 4 culture holes for experiment, and 5%
CO2, under the conditions of 37 DEG C act on 30 minutes after, remove culture fluid, wash with PBS three times, and add cell culture fluid with
The extract and medicine of variable concentrations is in 5%CO2, cultivate 72 hours under the conditions of 37 DEG C, with the antiviral of mtt assay Detection and Extraction thing
Activity.
4.2.3 the prophylactic activity of of the invention leaf of Flos Lonicerae extract, stem extract to virus multiplication：
Experiment is added to the extract of variable concentrations in Vero Tissue Culture Plates, 5%CO2, effect 8 is little under the conditions of 37 DEG C
Shi Hou, removes culture fluid, is washed with PBS three times, and each concentration extract sets 4 culture holes, and PEDV is viral for inoculation,
After acting on 30 minutes under the conditions of 37 DEG C, culture fluid is removed, washed with PBS three times, and add cell culture fluid 5%
CO2, cultivate 72 hours under the conditions of 37 DEG C, with prophylactic activity of the mtt assay Detection and Extraction thing to virus multiplication.
As shown in Table 3, 4, it is preferable that stem of Flos Lonicerae, leaf extract show in three kinds of model of action in vitro experimental result
Antiviral activity.
Inhibitory action (OD value) of the leaf of Flos Lonicerae extract of table 3 to PEDV cytopathogenic effects
Inhibitory action (OD value) of the stem of Flos Lonicerae extract of table 4 to PEDV cytopathogenic effects
Preventive and therapeutic effect of the stem and leaf of Flos Lonicerae feed additive of embodiment 5 to diarrhea of pigs
Pig feed experiment selects Semen Maydiss/bean cake/puffing Semen sojae atricolor type pig feed, Semen Maydiss, bean cake to be 1.8mm machinery sieve powder
It is broken.Its constituent is：It is Semen Maydiss 51.0%, puffing Semen sojae atricolor 16.2%, bean cake 13.0%, whey powder 6.4%, fish flour 6.2%, little
Testa Tritici 3.0%, stone powder 1.5%, dalcium biphosphate 1.0%, Sal 0.35%, lysine hydrochloride 0.22%, DL- protein acids
0.13%th, vitamin and microelement mixture 1.0%.Wherein, vitamin and microelement mixture contains in every 1 kilogram of feed
Zn180.0mg、Mn50.0mg、Se0.3mg、Fe150.0mg、Cu150.0mg、I 0.3mg、VD 20IU、VA 6500IU、VB1
2.8mg, VB2 6.2mg, VB6 1.1mg, VB12 35mg, VK3 3.5mg, nicotinic acid 33.0mg, Folic Acid 0.83mg, pantothenic acid
18.0mg etc..
Impact of the stem and leaf of Flos Lonicerae feed additive of 5.1 present invention to weaned piglets：
The using effect of stem and leaf of Flos Lonicerae feed additive prepared by the present embodiment research embodiment 2 in piglet, selects
Ju He plants health ablactational baby pig 250, statistical analysis body weight is randomly divided into 5 groups, per group 50 without significant difference by hero is female
Only.Experiment is carried out in slatted floor piglet nursing shed, and temperature control is 25～28 DEG C, and relative humidity is controlled at 50-70 DEG C.Piglet
Daily 8:0,12:00,16:00 and 20:00 timing feeding.Feedstuff mixes wet feeding manner using powder.And each forage volume
Slightly have clout for hopper to be advisable.Water is freely taken the photograph during experiment.House room is cleaned daily once.Every 4 days spraying disinfections once.Other
Carry out by pig farm feeding and management.
Record experiment starts with during terminating, and inventory and surpluses calculate body weight and feed intake.Test result indicate that,
With solvent control group, chlorogenic acid crude extract (>10%) and luteoloside crude extract (>10%) administration group is compared, containing 1.25% gold medal
Flos Lonicerae stem and leaf feed additive can improve piglet daily gain, reduce feedstuff score, promote cultivation quality and benefits.
Impact (kilogram) of the stem and leaf of Flos Lonicerae feed additive of the present invention of table 5 to weaned piglets
Preventive and therapeutic effect of the stem and leaf of Flos Lonicerae feed additive of 5.2 present invention to diarrhea of pigs：
Natural infection piglet 260, appetite is low, there is yellow-white loose stool.By daily early 7:30 to evening 21:30 continuous sights
Examine the defecation situation of piglet.Stools scored standard is：(1) 0 point：Bar shaped or particle shape, (2) 1 points：Soft excrement/shaping, (3) 2 points：It is thick
Shape/shaping/liquid dung without segregation phenomenon, (4) 3 points：Liquid/shapeless/liquid dung has segregation phenomenon.Abdomen is confirmed as by 2 points with 3 points
Rush down.Statistics piglet diarrhea number, calculates diarrhea rate.By laboratory diagnosiss, be diagnosed as yellow scour of piglet that escherichia coli cause and
Piglet pujos blancos.Sick pig is randomly divided into 5 groups by experiment, observes the phenomenons such as the feeding of each group, drinking-water, body weight, feces, and death.
Curative effect judging standard is that (1) cures：Sick pig diarrhoea disappears, and appetite is normal, without recurrence after being discontinued two weeks.(2) have
Effect：Diarrhoea mitigates or disappears, and fecal character takes a turn for the better, and moisture is significantly reduced, and the course of disease shortens.(3) it is invalid：Diarrhoea number of times and character without
Change, or the phenomena of mortality occur.Test result indicate that, with solvent control group, chlorogenic acid crude extract (>And luteoloside is thick 10%)
Extract (>10%) administration group is compared, and stem and leaf of Flos Lonicerae feed additive has on preventing and treating piglet yellow-white dysentery significantly to be controlled
Therapeutic effect.
Therapeutic effect of the stem and leaf of Flos Lonicerae feed additive of the present invention of table 6 to diarrhea of pigs
From the results show of table 6, the stem and leaf of Flos Lonicerae micropowder or extract of the present invention are added into after feedstuff, to pig abdomen
Rush down with significant therapeutic effect.Its therapeutic effect is more preferable compared with the chlorogenic acid of higher amount, luteoloside sterling effect.
The above is only the preferred embodiments of the present invention, and any pro forma restriction is not done to the present invention, though
So the present invention is disclosed above with preferred embodiment, but is not limited to the present invention, any to be familiar with this professional technology people
Member, in the range of without departing from technical solution of the present invention, when using the technology contents of the disclosure above make it is a little change or repair
The Equivalent embodiments for equivalent variations are adornd, as long as being the content without departing from technical solution of the present invention, according to the technology reality of the present invention
Any simple modification, equivalent variations and modification that confrontation above example is made, still fall within the scope of technical solution of the present invention
It is interior.
Claims (11)
1. it is a kind of preventing and treating Intestinum Sus domestica road transmission disease and improve efficiency of feed utilization feed additive, it is characterised in that the feeding
Feed additives raw material is made up of the stem or/and leaf of Flos Lonicerae；Gold containing percentage by weight 50～100% in the feed additive
Flos Lonicerae stem and leaf micropowder or extract.
2. it is according to claim 1 preventing and treating Intestinum Sus domestica road transmission disease and improve efficiency of feed utilization feed additive, its
It is characterised by, the feed additive contains 100% stem and leaf of Flos Lonicerae micropowder, the particle diameter of the stem and leaf of Flos Lonicerae micropowder powder is less than
0.85mm；Or stem and leaf of Flos Lonicerae water extract of the feed additive containing percentage by weight 50～100%, the stem of Flos Lonicerae
Per gram of leaf water extract contains crude drug>3g.
3. it is a kind of prepare described in claim 1 preventing and treating Intestinum Sus domestica road transmission disease and improve efficiency of feed utilization feed additive
Method, wherein, methods described includes：Powder by the raw material through mechanical activation comminution into particle diameter less than or equal to 0.85mm, obtains
To the feed additive.
4. it is a kind of prepare described in claim 1 preventing and treating Intestinum Sus domestica road transmission disease and improve efficiency of feed utilization feed additive
Method, wherein, methods described includes：
Step (1), by the raw material pulverizing into powder；
Step (2), by the step (1) obtain material powder addition solvent, the solvent be water, ethanol or aquiferous ethanol,
The solvent is water, ethanol or aquiferous ethanol, and the weight ratio of described material powder and solvent is 1:1～20, the solution of acquisition
The feed additive is obtained by heating extraction, microwave extraction or supersound extraction.
5. it is a kind of prepare described in claim 1 preventing and treating Intestinum Sus domestica road transmission disease and improve efficiency of feed utilization feed additive
Method, wherein, methods described includes：
Step (1), by the raw material pulverizing into powder；
Step (2), the material powder addition water that the step (1) is obtained, described material powder and the weight ratio of water is 1:1
～20, heating extraction, extraction time is 30 minutes to 24 hours, filters the extracting solution；
Step (3), by the filtering residue addition water after the filtration in the step (2), described filtering residue and the weight ratio of water is 1:1～
20, heating is extracted for second, and extraction time is 30 minutes to 24 hours, filters second extracting solution；
Step (4), merges the filtrate that the step (2) and step (3) are obtained, and is concentrated to give described feed additive.
6. method according to claim 5, wherein, heating-up temperature is boiling temperature in the step (2) and step (3),
Heat time heating time is 30 minutes.
7. it is a kind of preventing and treating Intestinum Sus domestica road transmission disease and improve efficiency of feed utilization feedstuff, it is characterised in that the feedstuff includes
Feed additive described in claim 1 of the weight than 1～5%.
8. feedstuff according to claim 7, it is characterised in that the feed additive is added by mixing with the feedstuff
Plus, or the feed additive is by being coated to the feed surface addition.
9. feed additive according to claim 1 is preparing the medicine of the prevention and treatment Intestinum Sus domestica road transmission disease
In application, it is characterised in that the Intestinum Sus domestica road transmission disease includes the popular diarrhoea of yellow and white dysentery of piglet, pig and pathogenic
Coli-infection or rotavirus infection and because of secondary digestive tract disease of suffering from diarrhoea.
10. application according to claim 9, it is characterised in that the infective pathogen of the Intestinum Sus domestica road transmission disease includes
Porcine epidemic diarrhea virus, transmissible gastro-enteritis viruss, rotavirus illness, Escherichia coli, staphylococcus aureuses,
Candida albicans, bacillus pyocyaneus, dysentery bacterium, enterobacteria.
11. feed additives according to claim 1 are improving Gaster Sus domestica intestinal microbial population balance, improve to feedstuff absorbability
With the application in terms of efficiency of feed utilization.
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| CN201610821184.7A CN106562037A (en) | 2016-09-13 | 2016-09-13 | Honeysuckle stem and leaf feed additive for preventing and treating pig intestinal tract infectious diseases and improving feed utilization ratio as well as preparation method and application thereof |
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| CN201610821184.7A CN106562037A (en) | 2016-09-13 | 2016-09-13 | Honeysuckle stem and leaf feed additive for preventing and treating pig intestinal tract infectious diseases and improving feed utilization ratio as well as preparation method and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107259159A (en) * | 2017-08-03 | 2017-10-20 | 湖南省产商品质量监督检验研究院 | Strengthen live pig immunity and fast well-fattened antimicrobial-free feed additive |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107259159A (en) * | 2017-08-03 | 2017-10-20 | 湖南省产商品质量监督检验研究院 | Strengthen live pig immunity and fast well-fattened antimicrobial-free feed additive |
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