CN106942521A - What a kind of property of medicine was stablized is used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations and method - Google Patents
What a kind of property of medicine was stablized is used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations and method Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
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Abstract
A kind of property of medicine it is stable be used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations and method, astragalus polyose 20% 50%, chlorogenic acid 20% 50%, allicin 20% 50%, sodium hydrogensulfite 5% 10%, the dimethyl beta propionic acid thialdine 3% 6% in terms of mass fraction.Its method includes pulverizing and sieving the Radix Astragali, mixes and decocts with distilled water, concentrates filtrate, plus ethanol, to alcohol content 80%, suction filtration, obtains astragalus polyose overnight；The bark of eucommia is pulverized and sieved, mixed with ethyl acetate, leaching is stirred at reflux in 65 DEG C of water-baths, merging filtrate isolates ethyl acetate, obtains chlorogenic acid.Preparation process of the present invention is simple; cost is relatively low; using convenient; prawn immune defense ability and bactericidal action can be effectively improved after said preparation production application; effectively preventing disease of prawn; the use of antibiotic and chemicals is reduced, technical support is provided for prawn culturing sustainable health development, technical guarantee is provided for water area ecological environment protection.
Description
Technical field
It is used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations and method the present invention relates to a kind of property of medicine stabilization, belongs to
Aquatic product matched field of feed additive technology.
Background technology
At present, prawn culturing turns into the pillar industry held the balance in culture fishery.With prawn culturing density and
The raising of intensive degree, causes disease to be on the rise by virus, bacterium etc., and causing tremendous economic to shrimp culture industry damages
Lose.For these diseases, medical treatment is most direct, most efficient method, still, chronically using antibiotic and other chemistry
Class medicine prevents and treats the farming disease harms, result in series of negative influence such as medicament residue, generation of drug-fast bacteria etc..With people couple
The use management and control of the more attention of ecological environment and human health, antibiotic and chemicals is increasingly strict, and people start to seek
Highly effective and safe approach improves the premunition of aquiculture animal.Chinese herbal medicine is not easily caused resistance to due to natural, efficient, low-residual
The advantages of property of medicine, the study hotspot of current aquiculture disease preventing and treating is had become, wherein compound Chinese herb is comprising a variety of effective
Composition, and each composition can act synergistically, and greatly enhance drug action, therefore compound Chinese herb has in terms of shrimp disease is prevented and treated
Have wide practical use.
The content of the invention
It is used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations it is an object of the invention to provide a kind of property of medicine stabilization
And method, by active component and other two kinds of compositions of three kinds of Chinese herbal medicines of extraction, collectively as prawn feed additive, both
Enhancing prawn immune defense ability and bactericidal action can be played, shrimp disease is effectively prevented and treated, antibiotic and chemicals is reduced
Use, can improve pharmacodynamic stability again and promotion is ingested, only prawn culturing sustainable health development does not provide technology branch
Support, also provides technical guarantee for water area ecological environment protection.
To reach above-mentioned purpose, the concrete technical scheme that the present invention takes is：
What a kind of property of medicine was stablized is used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations, it is characterised in that with mass fraction
Meter, it includes astragalus polyose 20%-50%, chlorogenic acid 20%-50%, allicin 20%-50%, sodium hydrogensulfite 5%-
10%th, dimethyl-β-propionic acid thialdine 3%-6%.
What a kind of property of medicine was stablized is used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations, it is characterised in that the Chinese herbal medicine system
Agent is consisted of the following composition, astragalus polyose：Chlorogenic acid：Allicin：Sodium hydrogensulfite：The mass ratio of dimethyl-β-propionic acid thialdine
For 14：14：7：2：1.
The above-mentioned formula be mainly used in it is daily feed, prawn immunocompetence can be significantly increased.
What a kind of property of medicine was stablized is used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations, it is characterised in that the Chinese herbal medicine system
Agent is consisted of the following composition, astragalus polyose：Chlorogenic acid：Allicin：Sodium hydrogensulfite：The mass ratio of dimethyl-β-propionic acid thialdine
For 14：14：14：2：1.
The above-mentioned formula is more suitable for feeding before the Growth season of prawn bacterial infection disease arrives, and such as cultivates
Middle and later periods, this stage daily ration, feeding quantity is continued to increase, water quality gradually aging, and residual bait and row's excreta increase, and pathogen quantity increases
Plus, shrimp ponds substrate deteriorates, and water quality is difficult to control to, and now disease easily breaks out.The formula has more significant biocidal efficacies.
Described astragalus polyose extracts from the Radix Astragali, and described chlorogenic acid extracts from the bark of eucommia, and astragalus polyose is pure heavy using water extraction
Method is extracted；Chlorogenic acid is extracted using ethyl acetate.Described allicin is chemical industry synthesis product, is had purchased from the high biotechnology of Jinan moral
Limit company；Described sodium hydrogensulfite is chemical industry synthesis product, purchased from Shandong Kai Long chemical science and technologies Development Co., Ltd：Described two
Methyl-β-propionic acid thialdine is chemical industry synthesis product, purchased from the chemical Science and Technology Ltd. of the rich bamboo grove in Wuhan.
The advantages of the present invention：
(1) Environment of Litopenaeus vannamei Low haemocyte quantity is improved.In prawn nonspecific immunity system, haemocyte plays important
Role, be both the executor and the supplier of humoral immunity of cellular immunity.Feeding Chinese herbal medicine immunopotentiator
In 21 days, test group prawn haemocyte quantity is all remarkably higher than control group (P<0.05), and control group is without significant difference.
(2) Environment of Litopenaeus vannamei Low haemocyte phagocytic activity is improved.Haemocyte can be wrapped up by phagocytosis, destroyed and eliminated
Foreign substance, therefore, haemocyte phagocytic activity can be used for weighing the ability that Environment of Litopenaeus vannamei Low resists exotic invasive pathogen.
Feed in 21 days of Chinese herbal medicine immunopotentiator, test group prawn haemocyte phagocytic activity is all remarkably higher than control group (P<
0.05), and control group is without significant difference.
(3) Environment of Litopenaeus vannamei Low Phenoloxidase Activities are improved.Feeding 21 days of Chinese herbal medicine immunopotentiator in, it is all
Shore prawn serum Phenoloxidase Activities of receiving are significantly improved, significant difference (P between each test group and control group<0.05).
(4) Serum of Penaeus vannamei antalzyme activity is improved.Lysozyme can destroy bacteria cell wall, be accordingly regarded as
Important humoral indications.Feeding 21 days of Chinese herbal medicine immunopotentiator in, Serum of Penaeus vannamei bacteriolyze enzyme activity
Power is significantly improved, significant difference (P between each test group and control group<0.05).
(5) resistivity of the Environment of Litopenaeus vannamei Low to morbid vibrio is improved.Inject 7d, control group prawn after Vibrio harveyi
Cumulative mortality be 68.35%, the cumulative mortality of the prawn of test group 1 is 33.35%, and the accumulation of the prawn of test group 2 is dead
Rate is 30%, significant difference (P between test group and control group<0.05).
(6) stability of Chinese herb compound drug effect is improved.Chlorogenic acid is the ester formed by caffeic acid and chinic acid, its point
There are ester bond, three l fractions of unsaturated double-bond and polyhydric phenols in minor structure, because of oxidation active ingredient can be caused to contain
Amount reduction.Meanwhile, allicin less stable, under the influence of external condition, color easily deepens even to become sticky, is divided quickly
Solve and produce a variety of diallyl sulfides, garlic fabricated product is produced obvious garlic odour taste and green and brown change, biological function and
Feeding effect is also decreased.How pre- anti-oxidation, it is to influence the key of chlorogenic acid and allicin to improve stable effective ingredients
Application factor.Current most of Chinese herb compounds on chlorogenic acid or allicin pertain only to the evaluation of effect, do not refer to it
The stability of drug effect.Sodium hydrogensulfite has strong reproducibility, in the present invention can be as antioxidant, and this patent is in formula
Middle addition sodium hydrogensulfite, increases the stability of Chinese herb compound, is conducive to the storage of Chinese herb compound.
(7) utilization rate of compound is improved.Chlorogenic acid has bitter taste, and allicin has pungent taste, and adding these compositions certainly will
Influence feed palatability.Dimethyl-β-propionic acid thialdine (DMPT) has good attractant effect to crustacean, multiple in Chinese herbal medicine
The DMPT of supplement low concentration can greatly promote prawn and ingest in side, improve the utilization rate of compound.
Experiment, which demonstrates immunopotentiator provided by the present invention, can be remarkably reinforced the immunity of prawn, improve prawn and support
The ability of anti-disease；The reinforcing agent effectively carries out scientific compatibility from astragalus polyose, chlorogenic acid, three kinds of Chinese herbal medicines of allicin, and
Coordinate the two class materials that can be improved pharmacodynamic stability and promote to ingest, be added in feed qualitative, quantitative, be conducive to maintaining
The stability and standardized production of drug effect；It can substitute and reduce the use of antibiotic and chemicals, it is to avoid antibiotic is indiscriminate
With the ill effect to environment etc., while the formula has higher promotion ration size；The present invention is prepared simply, and cost is relatively low, energy
Enough improve the economic benefit of shrimp culture industry.
Brief description of the drawings
Fig. 1 is the result figure of prawn haemocyte quantity in embodiment 2.
Fig. 2 is the result figure of prawn haemocyte phagocytic activity in embodiment 2.
Fig. 3 is the result figure of prawn blood plasma point oxidase active in embodiment 2.
Fig. 4 is the result figure of prawn blood plasma lysozyme activity in embodiment 2.
Fig. 5 is the result figure of challenge viral dosage prawn cumulative mortality in embodiment 2.
Embodiment
Embodiment 1：
The preparation method of Chinese herbal medicine immunopotentiator and basal feed, comprises the following steps：
Chinese herbal medicine active component is prepared first, after the sheet Radix Astragali is uniformly crushed, 80 mesh sieves is crossed, with distilled water according to quality
Volume ratio is 1:10 mix, and decoct 2 times, 2 hours every time, merge 2 filtrates, be concentrated under reduced pressure, plus appropriate 95% ethanol is so that most
Whole alcohol content is 80%, and overnight, suction filtration obtains astragalus polyose；
After the sheet bark of eucommia is uniformly crushed, 80 mesh sieves are crossed, with ethyl acetate (hydrochloric acid containing 0.05mol/L) according to quality volume
Than for 1:8 are mixed, and leaching 3 hours is stirred at reflux in 65 DEG C of water-baths, repeat to extract 2 times, merging filtrate isolates ethyl acetate,
Obtain chlorogenic acid；
Allicin, sodium hydrogensulfite and dimethyl-β-propionic acid thialdine use chemical industry synthesis product, buy high public from moral respectively
Department, the chemical Science and Technology Ltd. of the rich bamboo grove of Kai Long chemical science and technologies Development Co., Ltd and Wuhan；
Said herbal medicine extract, allicin, sodium hydrogensulfite and dimethyl-β-propionic acid thialdine is molten according to aforementioned proportion
Yu Shuihou is uniformly sprayed at basal feed surface, recycles fish oil parcel feed, dry obtain it is immune containing compound Chinese herb
The basal feed of reinforcing agent.
Reinforcing agent and corresponding breeding feed are prepared respectively according to above-mentioned preparation method, it is different by both the above
Specific proportioning obtains two kinds：In terms of mass fraction, a kind of immunopotentiator includes addition astragalus polyose 1g, green original per kilogram of feed
Sour 1g, allicin 0.5g, sodium hydrogensulfite 0.15g and dimethyl-β-propionic acid thialdine 0.07g；Another immunopotentiator includes
Addition astragalus polyose 1g, chlorogenic acid 1g, allicin 1g, sodium hydrogensulfite 0.15g and dimethyl-β-propionic acid thialdine per kilogram of feed
0.07g。
The method that the content of said herbal medicine active ingredient can be provided according to existing literature is measured.Astragalus polyose
The assay method reference of content：The measure of astragalus polyose content in the Radix Astragali, contemporary Chinese Chinese medicine, the 07th phase of volume 13 in 2011；
The assay method reference of chlorogenic acid content：Journal of Pharmaceutical Analysis, 2011 volume 1 the 04th
Phase；The assay method reference of Allicin Content：HPLC methods determine garlic in alliin and allicin content, 2009 volume 35
02nd phase；The measure of bisulfite sodium content uses oxidimetry；Dimethyl-β-propionic acid thialdine content uses sulphur weight
Method is detected.
Embodiment 2：
Influence of the Chinese herbal medicine immunopotentiator to Environment of Litopenaeus vannamei Low non-specific immunity
In this experiment, two kinds of Chinese medicine immunity enhancers that inventor prepares embodiment 1 feed Environment of Litopenaeus vannamei Low, to it
Salivary lysozyme is measured, it is intended to filter out raising Environment of Litopenaeus vannamei Low non-specific immune function and germ resistance energy
The Chinese herbal medicine immunopotentiator of power.
1 materials and methods
1.1 materials
Experiment is purchased from Qingdao City's Laoshan District sand mouthful prawn culturing with Environment of Litopenaeus vannamei Low (Litopenaeus vannamei)
Factory, average weight is 7.45 ± 0.41 grams, and experiment is with the composition of basal feed：In terms of mass fraction, fish meal 34%, dregs of beans
20%th, peanut meal 16.4%, wheat flour 22%, soybean lecithin 5%, multivitamin 0.6%, composite mineral matter 2%.Through experiment
Calculating nutritional ingredient in feed is：In terms of mass fraction, crude protein 42.4%, crude fat 7.2%, ash content 8.4%.
1.2 methods
1.2.1 experimental design
Experiment is randomly divided into 3 groups after temporarily being supported 7 days with Environment of Litopenaeus vannamei Low, respectively control group, test group 1 and test group 2, often
70 tails of group, every group 3 parallel, raises in PVC barrel of 100L, ventilation raising in 24 hours.Control group feeds basal feed, tests 1 He
2 groups of two kinds of immunopotentiator basal feeds feeding respectively prepared by embodiment 1, feed 2 times, are feeding volume of being satiated with food daily.
Culture experiment continues 21 days, during experiment altogether, water temperature, salinity, PH, respectively 25-27 DEG C, 34 ‰, 7.8-8.0.
1.2.2 testing index and method
In on-test the 0th, 1,3,6,10,15,21 days take blood.It is direct using No. 5 syringe needles and 1mL syringes of sterilization
3mm or so is taken a blood sample in insertion prawn carapace trailing edge cardiocoelom, and it is improved to have sucked 0.3mL in advance in the preceding syringe of blood drawing
Precooling Environment of Litopenaeus vannamei Low anti-coagulants (0.34M NaCl, 0.01M KCl, 0.01M EDTA-Na2,0.01M HEPES, pH 7.45,
Osmotic pressure is 780mOsm.kg-1), it is final so that the ratio of anti-coagulants and hemolymph is 1:1.A part of hemolymph sample enters promoting circulation of blood
The measure of cell quantity and phagocytic rate, remaining hemolymph sample is centrifuged after 10min in 800 × g at 4 DEG C, takes blue supernatant, is
Plasma sample, is preserved under the conditions of -80 DEG C.
The measure of haemocyte quantity is counted under an optical microscope using blood counting chamber；The measure side of phagocytic activity
Method is operated according to the method for existing literature, is referred to：Oxidative stress,DNA damage and antioxidant
enzyme gene expression in the Pacific white shrimp,Litopenaeus vannamei when
exposed to acute pH stress.Comparative Biochemistry and Physiology Part A；Phenol
Oxidase active assay method is referred to：White shrimp Litopenaeus vannamei that received
the hot-water extract of Gracilaria tenuistipitata showed earlier recovery in
immunity after a Vibrio alginolyticus injection.Fish&Shellfish Immunology；It is molten
Bacterium enzymatic activity builds up biological Co., Ltd using Nanjing and produces kit measurement.
2 results
2.1 haemocyte quantity
As shown in Figure 1：The feed containing Chinese herbal medicine immunopotentiator is fed after 21 days, Environment of Litopenaeus vannamei Low haemocyte quantity
It is significantly increased, after off-test, compared with control group, the haemocyte quantity of test group 1 and 2 increases by 14.2% He respectively
16.5%, test group and control group significant difference (p<0.05).But test group 2 is compared with test group 1, the rise of Allicin Content
Fail the quantity of further increase haemocyte.
2.2 haemocyte phagocytic activity
As shown in Figure 2：The feed containing Chinese herbal medicine immunopotentiator is fed after 6 days, the phagocytosis of Environment of Litopenaeus vannamei Low haemocyte
Rate starts to be significantly higher than control group, but the not notable (p of test group 1 and the difference of test group 2>0.05).In cultivation middle and later periods, test group
2 prawn haemocyte phagocytic activity increase is more obvious, is significantly higher than control group and test group 1, shows with culturing time
Extension, the allicin of high concentration is conducive to the phagocytic activity of further lifting Environment of Litopenaeus vannamei Low haemocyte.
2.3 phenoloxidase activities
As shown in Figure 3：The feed containing Chinese herbal medicine immunopotentiator is fed after 3 days, the oxidation of Serum of Penaeus vannamei phenol
Enzyme activity is significantly increased, and reaches peak in the 6th day test group 1 and the serum Phenoloxidase Activities of test group 2, with compareing
Group significant difference (p<0.05).From the 6th day to off-test, two test group serum Phenoloxidase Activities are declined slightly, but still are shown
Write and be higher than control group.The whole test period, test group 1 and the serum Phenoloxidase Activities difference of test group 2 is not notable.In summary
As a result it can obtain, astragalus polyose, chlorogenic acid and allicin are added in feed can improve Serum of Penaeus vannamei Phenoloxidase Activities, but
The further lifting of Allicin Content can not continue to strengthen prawn serum Phenoloxidase Activities.
2.4 lysozyme activities
As shown in Figure 4：The feed containing Chinese herbal medicine immunopotentiator is fed after 3 days, Serum of Penaeus vannamei lysozyme
Vigor is significantly increased, and reaches maximum at the 10th day, compared with control group, the increase of the serum lysozyme level of test group 1
2.05 times, the serum lysozyme level of test group 12 increases by 2.23 times, but phase each test group antalzyme activity has small after experiment
Amplitude declines and the significant difference (p between control group<0.05).
2.5 vibrios resistivities
As shown in Figure 5：Attack poison after 0.5 day each group prawn start dead, every group of death 1-3 tail occur, during by 1 day, compare
Group prawn adds up death toll apparently higher than each experimental group, during by 3 days, the reduction of each group death toll, after 5 days, each group prawn death toll
Reduce and tend towards stability.At the end of challenge viral dosage, control group cumulative mortality is up to 68.35%, the cumulative mortality of test group 1
For 33.35%, the cumulative mortality 30% of test group 2 shows that the Feed Energy for feeding addition astragalus polyose, chlorogenic acid and allicin is carried
The high anti-vibrios ability of Environment of Litopenaeus vannamei Low, and anti-vibrios ability improves with the increase of Allicin Content.
Claims (5)
- What 1. a kind of property of medicine was stablized is used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations, it is characterised in that with mass fraction Meter, it includes astragalus polyose 20%-50%, chlorogenic acid 20%-50%, allicin 20%-50%, sodium hydrogensulfite 5%- 10%th, dimethyl-β-propionic acid thialdine 3%-6%.
- What 2. a kind of property of medicine was stable as claimed in claim 1 is used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations, its spy It is that the Chinese herbal and crude drugs preparations is consisted of the following composition to levy, astragalus polyose：Chlorogenic acid：Allicin：Sodium hydrogensulfite：Dimethyl-β- The mass ratio of propionic acid thialdine is 14：14：7：2：1.
- What 3. a kind of property of medicine was stable as claimed in claim 1 is used for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations, its spy It is that the Chinese herbal and crude drugs preparations is consisted of the following composition to levy, astragalus polyose：Chlorogenic acid：Allicin：Sodium hydrogensulfite：Dimethyl-β- The mass ratio of propionic acid thialdine is 14：14：14：2：1.
- 4. a kind of stable preparation method for prawn Immune-enhancing effect and sterilized Chinese herbal and crude drugs preparations of property of medicine, it is characterised in that bag Include following steps：Chinese herbal medicine active component is prepared first, after the sheet Radix Astragali is uniformly crushed, 80 mesh sieves is crossed, with distilled water according to quality volume Than for 1:10 mix, and decoct 2 times, 2 hours every time, merge 2 filtrates, be concentrated under reduced pressure, plus appropriate 95% ethanol is so as to finally contain Alcohol amount is 80%, and overnight, suction filtration obtains astragalus polyose；After the sheet bark of eucommia is uniformly crushed, 80 mesh sieves are crossed, are according to mass volume ratio with ethyl acetate (hydrochloric acid containing 0.05mol/L) 1:8 are mixed, and leaching 3 hours is stirred at reflux in 65 DEG C of water-baths, are repeated to extract 2 times, merging filtrate isolates ethyl acetate, are obtained Chlorogenic acid；Allicin, sodium hydrogensulfite and dimethyl-β-propionic acid thialdine use chemical industry synthesis product,By above-mentioned astragalus polyose, chlorogenic acid, allicin, sodium hydrogensulfite, dimethyl-β-propionic acid thialdine according to claim 1 or 2 Or the ratio described in 3 is matched, and is produced.
- 5. a kind of preparation method of the basal feed containing Chinese herbal medicine immunopotentiator, it is characterised in that including following step Suddenly：Astragalus polyose, chlorogenic acid, allicin, sodium hydrogensulfite, dimethyl-β-the third are obtained in accordance with the method for claim 4 Sour thialdine, by above-mentioned astragalus polyose, chlorogenic acid, allicin, sodium hydrogensulfite, dimethyl-β-propionic acid thialdine according to claim 1 Or the ratio described in 2 or 3 is dissolved in after water and is uniformly sprayed at basal feed surface, fish oil parcel feed is recycled, dries and obtains A kind of basal feed containing Chinese herbal medicine immunopotentiator.
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| CN106942521B (en) | 2020-09-15 |
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