CN103919889B - A kind of Chinese medicine composition strengthening turbot immunity - Google Patents
A kind of Chinese medicine composition strengthening turbot immunity Download PDFInfo
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- CN103919889B CN103919889B CN201410181408.3A CN201410181408A CN103919889B CN 103919889 B CN103919889 B CN 103919889B CN 201410181408 A CN201410181408 A CN 201410181408A CN 103919889 B CN103919889 B CN 103919889B
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Abstract
The invention discloses a kind of Chinese medicine composition for improving turbot immunity and preparation method thereof, belonging to marine fishes technical field of pharmaceuticals.It is characterized in that forming according to certain proportion proportioning with Echinacea, Poria, Herba Hedyotidis Diffusae, Radix Isatidis.The turbot of medicinal application of the present invention after giving Vibrio anguillarum vaccine fundamental immunity, significantly can strengthen the immunity of turbot, thus effectively reduces the death of Fish, reduces the loss of turbot cultivation.
Description
Technical field
The invention belongs to marine fishes technical field of pharmaceuticals, be specifically related to a kind ofly strengthen Chinese medicine composition of turbot immunity and preparation method thereof.
Background technology
Turbot is under the jurisdiction of Pleuronectiformes, Bothidae, and brill belongs to, and is seawater fish of dwelling at the bottom of a kind of cold water, originates in the marine sites such as northeast, the Atlantic Ocean, the North Sea, Mediterranean and the Baltic Sea.Turbot is good because having meat, is of high nutritive value, and growth is fast, is applicable to the life of low water temperature, and culture benefit is high and deeply the advantage such as to like by consumer and in succession become one of high-quality marine fish developed various countries.Although the artificial Isolated from Diseased Scophthalmus maximus of China 4-5 more late than Europe, the annual production of China's turbot in recent years has reached annual more than 20,000 tons, and annual value of production breaks through 2,000,000,000, is one of main sea-farming kind of Northern Part of China.But along with the cultivation expansion of scale and the deterioration of water body environment, the harm that various disease causes is also gradually by raiser and fisheries expert are paid attention to.Wherein serious with bacterial disease harm.The Vibrios such as Vibrio anguillarum (Vibrioanguillarum), Vibro harveyi (Vibrioharvey), Vibrio splindidus (Vibriosplendidus), Fermi operator (Vibriofischeri) are mainly comprised sick in the turbot bacterial pathogen of domestic and international report.In vibriosis, most is representational is that Vibrio anguillarum is sick, and its virulence is the strongest, can cause the death of large-scale turbot seedling and adult fish.
Vibrio anguillarum endangers maximum pathogen in turbot pathogen, worldwide popular.Vibrio anguillarum is widely distributed at occurring in nature, is the normal flora in water environment, and when water temperature better nutritivity composition enriches very much, its quantity can sharply increase, and can cause the epidemic disease of aquatic animal particularly Fish.Vibrio anguillarum can cause the acute sepsis of adult turbot and fry, finally causes the massive mortality of disease fish.It is mainly by skin, the gill, side line and intestinal infection Fish, and the most obvious symptom of metainfective turbot is that body surface festers and fin severe haemorrhage, and sick fish is usually dead in 2-3 days.
And in current aquaculture, with treatment, various chemicals and antibiotic are importantly used to the prevention of fish disease.The medium-term and long-term medication of fish culture process makes pathogenic bacteria drug resistance strengthen, and a large amount of antibiotic use also can make the immunocompetence of Fish decline, and in aquatic products, residual medicine also has a strong impact on the safety of food and the health of the mankind simultaneously.The aeromonas salmonicida oral immunity rainbow trout (Salmo gairdneri) of Duff (1942) first Application deactivation succeeds, thus has started vaccine new era in Fish application.After entering the seventies, various immune formulation is developed in succession, such as Vibrio anguillarum vaccine, grass carp hemorrhage disease vaccine, Trionyx sinensis Wiegmann bacterial disease vaccine etc.Current vaccines for fish kind is more, but because some vaccine is because of the particularity of its preparation method or pathogen, causes its less immunogenic, and after immunity, protected effect is not good.Therefore be need to coordinate efficient adjuvant to use when developing vaccine.
Application Chinese herbal medicine not only can solve anti-(resistance to) property of medicine that chemicals, antibiotic etc. cause and the residual problem such as to exceed standard of cultured fishes medicine as immunostimulant, and meets development non-public-hazard fishery, produces the demand of green aquatic product.Lot of domestic and international scholar's research proves, Chinese medicine immunity enhancer obviously can strengthen the resistance against diseases of aquatic animal.If any research astragalus polysaccharides, the compound recipe such as Radix Achyranthis Bidentatae, Radix Isatidis, saponin etc. have enhancing immunization to turbot, but do not have the Vibrio anguillarum vaccine for turbot should be used for improving the report of immunity.
Summary of the invention
The object of this invention is to provide a kind of Chinese medicine composition strengthening turbot immunity, and the preparation method of said composition, Chinese medicine composition clinical application effect prepared by the present invention be good, cost is low, pure Chinese medicinal preparation, noresidue, there is good application prospect.
The present invention strengthens the Chinese medicine composition of turbot immunity, comprises the crude drug of following weight portion: Echinacea 80 ~ 120 parts, 40 ~ 60 parts, Poria, Herba Hedyotidis Diffusae 30 ~ 50 parts, Radix Isatidis 30 ~ 50 parts.
Above-mentioned Chinese medicine composition is for the preparation of the adjuvant for strengthening turbot immunity;
Described adjuvant is solid or liquid condition,
The preparation method of solid forms is by Echinacea, Poria, Herba Hedyotidis Diffusae, Radix Isatidis, is ground into fine powder and obtains;
The another kind of preparation method of the adjuvant of solid forms gets Echinacea, Poria, Herba Hedyotidis Diffusae, Radix Isatidis, adds 12 times of water gagings, ebuillition of heated extracts 2 times, each 1.5 hours, filters, measuring relative density during filtrate reduced in volume to 60 DEG C is 1.30 ~ 1.35, and drying under reduced pressure is pulverized obtained;
The preparation method of liquid form: get Echinacea, Poria, Herba Hedyotidis Diffusae, Radix Isatidis, add 12 times of water gagings, ebuillition of heated extracts 2 times, each 1.5 hours, filters, and measuring relative density during filtrate reduced in volume to 60 DEG C is 1.10 ~ 1.15, filters obtained:
In practical application, can fine medicinal material powder, extract fine powder or extracting solution be selected to combine the application of Vibrio anguillarum vaccine according to different situations respectively.Injection, immersion, the mode administration such as oral can be adopted.
Chinese medicine composition of the present invention is mainly used in the application of associating Vibrio anguillarum vaccine, to improve the immunity of turbot
Chinese medicine composition of the present invention has selected Echinacea, Poria, Herba Hedyotidis Diffusae, Radix Isatidis four Chinese medicine material, is mainly used in associating Vibrio anguillarum vaccine to improve the immunity of turbot.After Chinese medicine composition of the present invention and the coupling of Vibrio anguillarum vaccine, the immunity of turbot can be strengthened, the more alone Chinese medicine composition of its immune effect or Vibrio anguillarum vaccine good.
Detailed description of the invention
Echinacea medical material in Chinese medicine composition of the present invention, containing plurality of active ingredients such as polysaccharide, flavone, caffeinic derivant, quintessence oil, polyacetylene, alkylamine and alkaloids, has immunostimulation, reduces inflammation, can to antibacterium and virus.Experiment proves Echinacea stimulating immune system cellular activity.When these cells have activity, leukocyte causing the invader of disease to surround, making virus or bacterial death, preventing them from breeding, or only stopping their activity more rapidly.As far back as 17th century, U.S. original inhabitant just uses Echinacea, and they remove to treat the various diseases such as disease, flu, cough, septicemia, sore throat, gastroenteralgia in venom, gingiva and oral cavity by this kind of plant.
Poria medical material, is the sclerotium of On Polyporaceae Poria, comes from Shennong's Herbal.Ancients claim Poria to be " when four god medicine ", because its effect widely, regardless of the four seasons, by it and various compatibility of drugs, no matter cold, temperature, wind, wet all diseases, can play its unique effects.Poria sweet in the mouth, light, property are put down, and are used as medicine and have the function of promoting diuresis to eliminate damp pathogen, strengthening the spleen stomach function regulating, mind tranquilizing and the heart calming.Modern medicine study shows, Poria energy enhancing human body immunity function, pachyman has obvious antitumor and protects the liver dirty effect.Effect of latest find shows, the main component in Poria is pachyman, and content is very high.Inhibitory action is had to various bacteria; Can gastric acid be fallen, have preventive effect to digestive tract ulcer; Significant protective effect is had to hepatic injury; There is antineoplastic action; Many-sided immunologic function can be regulated; Leukopenia caused by chemotherapy can be made to accelerate; And have calm effect.
Herba Hedyotidis Diffusae medical material is bitter in the mouth, light, cold in nature.Cure mainly: dyspnea and cough due to lung-heat; Laryngopharynx swelling and pain; Intestinal carbuncle; Furuncle and phyma skin infection; Venom; The puckery pain of pyretic stranguria; Edema; Dysentery; Intestinal is scorching; Jaundice due to damp-heat; Be good at the multiple cancerous protuberance for the treatment of.Herba Hedyotidis Diffusae strengthens immunologic function successful, in vitro tests shows that this product rear anti-infectious function for oral administration is obvious, be by stimulating reticuloendothelial system, increase phagocytic activity of leukocytes to improve serum sterilizing power, the ability of administration group leukocytes phagocytic Congo red comparatively matched group improves 4 times.The remarkable hypertrophy of the visible reticuloendothelial system of microscopy, skein cell hypertrophy, endochylema enriches, and engulfs active, and in the tissue such as lymph node, spleen, liver, argentophile substance is that densification changes, and display body defense function is strengthened.Can humoral immune function be strengthened, hemolysis plaque digital display work in mouse spleen is increased.
Chromatogram of Radix Isatidis, bitter in the mouth is cold in nature, GUIXIN, stomach, liver, gallbladder meridian.Function heat-clearing and toxic substances removing, removing heat from blood sore-throat relieving.Cure mainly the disease caused by warm poison, as influenza, upper respiratory tract infection, epidemic encephalitis, parotitis, acute enteritis, bacillary dysentery, hepatitis, facial erysipelas, calentura send out speckle etc.Pharmacological research shows, Radix Isatidis has obvious inhibitory action to multiple virus and pathogenic bacteria, Radix Isatidis has external broad-spectrum antibacterial action (comprising dysentery antibacterial, salmonella bacteria, hemolytic rope-forming bacteria), be anti-inflammation, pain relieving, the Chinese traditional herbs of bringing down a fever.After its antiviral, anti-inflammation, enhance the immunity of body simultaneously.
The present invention, by long-term research, by filtering out four kinds of above-mentioned components and proportioning to the detection of turbot immune effect, thus facilitates the present invention.
With specific embodiment, Chinese medicine composition prepared by the present invention is described again below, following examples are only for illustration of the present invention, and the present invention is not limited to this.
Embodiment 1
Prescription: Echinacea 80g, Poria 40g, Herba Hedyotidis Diffusae 30g, Radix Isatidis 30g
Method for making: get above-mentioned each medical material, be ground into fine powder, sieve, subpackage every bag 500g, to obtain final product.
Embodiment 2:
Prescription: Echinacea 500g, Poria 250g, Herba Hedyotidis Diffusae 200g, Radix Isatidis 200g
Method for making: get above-mentioned each medical material, add 13.8L water, ebuillition of heated extracts 2 times, each 1.5 hours, filter, measuring relative density during filtrate reduced in volume to 60 DEG C is 1.30 ~ 1.35, drying under reduced pressure, and pulverize, subpackage every bag 500g, to obtain final product.
Embodiment 3:
Prescription: Echinacea 12kg, Poria 6kg, Herba Hedyotidis Diffusae 5kg, Radix Isatidis 5kg
Method for making: get above-mentioned each medical material, add 336L water, ebuillition of heated extracts 2 times, each 1.5 hours, filter, measuring relative density during filtrate reduced in volume to 60 DEG C is 1.10 ~ 1.15, and filter, fill, every bottle of 500ml, to obtain final product.
The immune effect of Chinese medicine composition of the present invention to the turbot after fundamental immunity Vibrio anguillarum vaccine compares
(1) laboratory animal
Choose turbot 300 tail, be divided into 5 groups.Turbot body is long is 7.5-9.5cm, body weight 8.0 ± 1.5g.Supporting temporarily in cement pit before experiment is placed in the circular glass steel drum of the diameter 1.0m of customization for 2 weeks, flowing water culture.Breeding water is that fresh water and underground sea water are blent, constant temperature 18 ± 1 DEG C, salinity 32, pH8.0.Remove contamination every day 2 times, throw something and feed 3 times, daily ration of feeding accounts for 1.5% of weight.
(2) Experimental agents
Chinese medicine of the present invention: the medicated powder in Example 2, get 1g add water 150ml dissolve;
Vibrio anguillarum vaccine: adopt commercially available Vibrio anguillarum vaccine (be qualified vaccine through steriling test and safety examination).
(3) experiment content
Immunity test: be divided into 5 groups at random to turbot in experiment, often organizes 60 tail fishes, adopts infusion method to carry out immunity, is respectively medicine group of the present invention, Vibrio anguillarum vaccine group, medicine of the present invention+Vibrio anguillarum vaccine group, astragalus polysaccharides+Vibrio anguillarum vaccine group and sea water group.First by each group with corresponding drug immersion 10min, then drain, add vaccine carry out immersion 30min (vaccine concentration 1 × 108CFU/ml, without vaccine group second time with sea water immersion, with sea water group for contrast).
Counteracting toxic substances infects to be tested with immune relative protection ratio: exempted from fish be divided into 3 groups at random by often organizing, often organize 20 tails, respectively at immunity the 7th, 15, after 30d, carry out immersion counteracting toxic substances with Vibrio anguillarum MN, counteracting toxic substances bacteria concentration is according to the result of LD50.Dissect dead turbot and also carry out pathogenicbacteria separation and qualification, to confirm death whether caused by infection pathogen.Within after counteracting toxic substances 14 days, record death condition day by day, Computation immunity relative protection ratio (RPS).
Experimental group data acquisition SPSS12.0 software carries out statistical analysis.
(3) experimental result
1. the cumulative mortality respectively organized after immunity 7 days counteracting toxic substances and RPS
Within 7th day, carry out first time counteracting toxic substances in immunity, adopt the mode of soaking counteracting toxic substances.The concentration of counteracting toxic substances Vibrio anguillarum used is 15 times of LD
50dosage, infects 12 days, record each group every day cumulative mortality, and calculate RPS.The results are shown in Table 1.
The immunity of table 1 turbot respectively organizes mortality rate and RPS in 7 days afterwards
Note:
*p, compares with sea water group;
△p, compares with Vibrio anguillarum vaccine group;
#p, compares with astragalus polysaccharides group
2. the cumulative mortality respectively organized after immunity 15 days counteracting toxic substances and RPS
Within 15th day, carry out second time counteracting toxic substances in immunity, adopt the mode of immersion counteracting toxic substances, infect 12 days, record each group every day cumulative mortality, and calculate RPS.The results are shown in Table 2.
The immunity of table 2 turbot respectively organizes mortality rate and RPS in 15 days afterwards
Note:
*p, compares with sea water group;
△p, compares with Vibrio anguillarum vaccine group;
#p, compares with astragalus polysaccharides group
3. the cumulative mortality respectively organized after immunity 30 days counteracting toxic substances and RPS
Within 30th day, carry out third time counteracting toxic substances in immunity, adopt the mode of immersion counteracting toxic substances, infect 12 days, record each group every day cumulative mortality, and calculate RPS.The results are shown in Table 3.
The immunity of table 3 turbot respectively organizes mortality rate and RPS in 30 days afterwards
Note:
*p, compares with sea water group;
△p, compares with Vibrio anguillarum vaccine group;
#p, compares with astragalus polysaccharides group
Result shows, medicine of the present invention+Vibrio anguillarum vaccine group can significantly improve the immunity of turbot, compares have pole significant difference with blank group; Compare with alone Vibrio anguillarum vaccine group, there is significant difference; Chinese medicine composition of the present invention, after the coupling of Vibrio anguillarum vaccine, after 30 days, still there is good immunity in turbot immunity, and astragalus polysaccharides group declines fast in the immunity of turbot immunity after 30 days, compare with bibliographical information effect good astragalus polysaccharides+Vibrio anguillarum vaccine group, there is significant difference.And, in the process using vaccine of the present invention, the feedstuff picked-up of turbot can not be reduced, the growth rate of turbot can not be affected when enhancing immunity.Visible, medicine of the present invention has higher using value, has good competitiveness in Fish industry.Medicinal application of the present invention has a extensive future.
Embodiment 4: practical application effect
There is turbot fry 3.5 ten thousand tail fish culture field, Huangdao District, Qingdao of Shandong province.Wherein 2.1 ten thousand tails give the immunity of the medicine in Vibrio anguillarum vaccine and embodiment 3 respectively, and 0.4 ten thousand tails in another fishpond, only give Vibrio anguillarum vaccine immunity because medicine of the present invention is finished.As a result, in 30 days, the turbot cumulative mortality giving medicine of the present invention and Vibrio anguillarum vaccine immunity is 13.4%, and the turbot cumulative mortality only giving Vibrio anguillarum vaccine immunity is 35.7%.
Show that Chinese medicine composition prepared by the present invention effectively can improve the immunity of turbot through multiple embodiment, instead of because the case that causes of the difference on individual or Therapeutic Method.Chinese medicine composition of the present invention has good application and popularization value.
Claims (6)
1. for strengthening a Chinese medicine composition for turbot immunity, it is characterized in that, described Chinese medicine composition is made up of the crude drug of following weight portion: Echinacea 80 ~ 120 parts, 40 ~ 60 parts, Poria, Herba Hedyotidis Diffusae 30 ~ 50 parts, Radix Isatidis 30 ~ 50 parts.
2. for strengthening an adjuvant for turbot immunity, it is characterized in that, prepared by described adjuvant Chinese medicine composition according to claim 1.
3. adjuvant as claimed in claim 2, it is characterized in that, described adjuvant is pressed powder form, is by the Echinacea in Chinese medicine composition described in claim 1, Poria, Herba Hedyotidis Diffusae, Radix Isatidis, is ground into fine powder obtained.
4. adjuvant as claimed in claim 2, it is characterized in that, described adjuvant is solid forms, that the Echinacea in Chinese medicine composition described in claim 1, Poria, Herba Hedyotidis Diffusae, Radix Isatidis are added 12 times of water gagings, ebuillition of heated extracts 2 times, each 1.5 hours, filters, measuring relative density during filtrate reduced in volume to 60 DEG C is 1.30 ~ 1.35, and drying under reduced pressure is pulverized obtained.
5. adjuvant as claimed in claim 2, it is characterized in that, described adjuvant is liquid form, that the Echinacea in the described Chinese medicine composition of claim 1, Poria, Herba Hedyotidis Diffusae, Radix Isatidis are added 12 times of water gagings, ebuillition of heated extracts 2 times, each 1.5 hours, filters, measuring relative density during filtrate reduced in volume to 60 DEG C is 1.10 ~ 1.15, and filtration obtains.
6. improve a preparation for turbot immunity, it is characterized in that, described preparation is that adjuvant according to claim 2 combines preparation with Vibrio anguillarum vaccine.
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