CN115414378A - Application of immunopotentiator based on scutellaria baicalensis polysaccharide in eriocheir sinensis breeding - Google Patents
Application of immunopotentiator based on scutellaria baicalensis polysaccharide in eriocheir sinensis breeding Download PDFInfo
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- CN115414378A CN115414378A CN202210677322.4A CN202210677322A CN115414378A CN 115414378 A CN115414378 A CN 115414378A CN 202210677322 A CN202210677322 A CN 202210677322A CN 115414378 A CN115414378 A CN 115414378A
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- Prior art keywords
- polysaccharide
- scutellaria
- eriocheir sinensis
- immunopotentiator
- bait
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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- 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)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
- A61K36/539—Scutellaria (skullcap)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- General Health & Medical Sciences (AREA)
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Abstract
The invention discloses a preparation process of bait taking scutellaria baicalensis polysaccharide as an immunopotentiator and application of the bait in eriocheir sinensis cultivation. The prepared baical skullcap root polysaccharide can obviously improve the inherent immunity of the eriocheir sinensis, and comprises the steps of obviously inhibiting the proliferation of aeromonas hydrophila of eriocheir sinensis pathogenic bacteria and improving the survival rate of the eriocheir sinensis for resisting the infection of the aeromonas hydrophila; remarkably improves the total number of the eriocheir sinensis hemolymph cells, enhances the phagocytic activity of the hemolymph cells and inhibits the apoptosis of the hemolymph cells; obviously improve the expression quantity of immune effector molecules such as lysozyme, peroxidase and the like in the eriocheir sinensis hemolymph serum and inhibit the generation of inflammatory cytokines. The baicalein used in the invention is derived from organic wastewater generated in the baicalin extraction process, and the content of the baicalein in the wastewater is about 5.20 percent, so the invention has the characteristics of low cost, no toxic or side effect, remarkable antibacterial effect and immunity enhancement effect, improves the utilization rate of the baicalin and reduces the environmental pollution.
Description
Technical Field
The invention relates to a baicalein derived from baicalin extraction waste liquid and application thereof as an immunopotentiator in eriocheir sinensis baits.
Background
Chinese mitten crabs (Eriocheir sinensis), also called hairy crabs, river crabs, hairy crabs or clean water crabs, belong to the phylum Arthropod, crustacea, decapoda, archaedae and Eriocheir sinensis, and are important famous and special-quality fresh water economic breeding varieties in China. At present, eriocheir sinensis is cultivated all over the country (except for autonomous region in Tibet), wherein the yield of 6 provinces of Jiangsu, hubei, anhui, liaoning, jiangxi and Shandong accounts for about 93 percent of the total yield of the country. According to the report of 'Chinese fishery statistics yearbook', the total output of Chinese eriocheir sinensis in 2018 is as high as 75 ten thousand tons. However, with the continuous expansion of the culture scale, various diseases of the eriocheir sinensis frequently occur, which causes great loss to the river crab culture industry and seriously restricts the sustainable development of the river crab culture industry.
Antibiotics and chemical disinfectants are still the main prevention and control measures for preventing and controlling the diseases of the river crabs so far, but the prevention and control method not only pollutes the water body environment and destroys the diversity of aquatic organisms, but also can cause the generation of drug-resistant bacteria. Therefore, in the whole aquaculture industry, a prevention and treatment technology which is environment-friendly, has an obvious prevention and treatment effect and small toxic and side effects and can enhance the body immunity of the river crabs is urgently needed to be found.
The traditional Chinese medicine polysaccharide is a broad-spectrum and high-efficiency immunomodulator, and is an immunopotentiator which is most widely applied and deeply researched in the field of aquaculture. However, in the Eriocheir sinensis breeding industry, related products of the traditional Chinese medicine polysaccharide immunopotentiator are not yet seen on the market. The invention provides technical support for the development of the river crab immunopotentiator product and promotes the development and popularization of the river crab immunopotentiator product.
Disclosure of Invention
The invention aims at one: the method is comprehensive utilization of waste liquid in the baicalin extraction process, prolongs the product line of the baicalin extraction process, and reduces the content of organic matters in the baicalin extraction waste liquid; simultaneously, rapidly recovering and drying the scutellaria baicalensis polysaccharide recovered from the waste liquid;
object two of the present invention: provides a preparation method of bait based on scutellaria polysaccharide as an immunopotentiator, and definitely determines the application of the bait in the eriocheir sinensis breeding industry.
The third purpose of the invention is that: the immunity enhancing mechanism of the scutellaria baicalensis polysaccharide to the Eriocheir sinensis is determined, and the mechanism comprises antibacterial activity and the regulating effect of the scutellaria baicalensis polysaccharide on innate immunity.
The invention adopts the following technical scheme:
(1) And (3) waste liquid treatment: the baicalin extraction waste liquid is a strongly acidic solution, the baicalin is completely retained in the waste liquid, and in order to prevent the waste liquid from corroding subsequent polysaccharide extraction equipment, the pH value is firstly adjusted to be weakly acidic or neutral;
(2) And (3) concentrating: in the process of extracting the scutellaria baicalensis, after multiple filtrates are combined, the generated waste liquid is about more than 30 times of the dry weight of the scutellaria baicalensis, according to the content of the scutellaria baicalensis polysaccharide, weakly acidic or neutral waste liquid needs to be concentrated, and the waste liquid is heated in a water bath and is subjected to reflux concentration;
(3) Extraction: adding 3-4 times of edible alcohol by conventional ethanol precipitation method to obtain Scutellariae radix polysaccharide precipitate;
(4) And (3) filter pressing: discarding supernatant, and pressing Scutellariae radix polysaccharide into block solid by filter pressing method;
(5) And (3) quick drying: slicing the block-shaped radix Scutellariae polysaccharide with a slicer, and placing in an oven to obtain dried radix Scutellariae polysaccharide;
(6) The formula design is as follows: the eriocheir sinensis bait with good stability in water and remarkable immunity enhancing effect is prepared by taking fish meal, river snail meat powder, peanut cake powder, rice flour starch and the like as basic nutritional formulas and taking scutellaria polysaccharide as an immunopotentiator.
The pH value in the step (1) is between 6.0 and 7.0;
in the step (2), the concentration temperature is 50-70 ℃, the concentration vacuum degree is 0.4-0.8 Mpa, and the volume of the concentrated solution is about 10-30% of the total volume before concentration;
in the step (3), the edible alcoholic strength is more than 90% (v/v), the alcoholic strength in the mixed solution of the alcohol and the baical skullcap root polysaccharide is 70-80% (v/v), the precipitation time is 3-8 hours, and the precipitation temperature is 4-10 ℃;
in the step (4), the water content in the massive solid scutellaria baicalensis polysaccharide is between 15 and 35 percent;
in the step (5), the thickness of the scutellaria baicalensis polysaccharide sheet is about 0.5-2 mm, the temperature of the oven is set to be 40-60 ℃, and the drying time is 0.5-3 h;
in the step (6), the content of the scutellaria baicalensis polysaccharide in the river crab bait accounts for about 1.0-3.0%, and the stability of the bait in water is more than 2 hours.
Preparation of scutellaria polysaccharide coated pellet for river crab bait
Preparation of basal pellets
The scutellaria polysaccharide river crab bait comprises the following components: 38% of white fish meal, 15% of river snail meat powder, 10% of peanut cake powder, 5% of bean cake powder, 4% of sweet corn flour, 3% of bone meal, 6% of green fish meal, 17% of shrimp meat powder and 2% of astragalus polysaccharide. Pouring the substances into a grinder according to a certain proportion, grinding, taking out, putting into a stirrer, stirring for 30 minutes, uniformly mixing, adding purified water, continuously stirring to prepare a wet material which is 'lightly held into a mass and is immediately scattered by contact', putting into an extruder, and adjusting the extrusion speed to 200 r.min -1 Extruding the mixture through a sieve plate (with the aperture of 1.5 mm) of an extruder, putting the strip-shaped material into a spheronizer, and carrying out spheronization at the rotating speed of 400 r.min < -1 > for 4 min so as to ensure that the particles are completely spheronized. Taking out the pellets, drying, carrying out heat treatment, and screening out 12-30 meshes of pellets for later use, wherein the yield is 94.3%.
Coating of basal pellets
Adding 5% (w/v) of talcum powder into the ethyl cellulose water dispersion (25%), stirring the purified water with the same volume uniformly, homogenizing the mixture for 3min by using a high-speed dispersion homogenizer, and sieving the homogenized mixture by using a 100-mesh sieve to obtain a coating solution, wherein the coating solution is continuously stirred in the coating process, and the coating weight is increased by 10%. Weighing 1000g of base pills, preheating the base pills at 30 ℃ for 3min, placing the base pills in a fluidized bed coating machine, and adopting bottom spraying, wherein the spraying speed is 1.0-1.5 mL/min < -1 >, the air inlet frequency is 30-40 Hz, and the temperature in the fluidized bed coating machine is always kept at 25-30 ℃. Keeping the pellets from being bonded in the operation process, mixing the pellets with 0.2% (w) of micropowder silica gel after coating, drying in a 50 ℃ oven, measuring the moisture of the pellets by using a moisture meter until the moisture is less than or equal to 1%, and collecting the coated pellets of 12-30 meshes.
Measurement of Baikal skullcap root polysaccharide Release degree
The second method of Chinese pharmacopoeia 2020 annual edition release degree determination (0931 dissolution and release degree determination), taking scutellaria polysaccharide river crab bait coating pellet (about equivalent to 1.0g of astragalus polysaccharide), respectively taking 1000ml of sodium phosphate buffer solution (pH8.0), phosphate buffer solution (pH6.8) and acetate buffer solution (pH4.0) as release medium, rotating speed is 75 r/min, operating according to the method, taking 5ml of solution 2h, filtering, taking subsequent filtrate, and determining absorbance A1 at 490nm wavelength; grinding Scutellariae radix polysaccharide coated pellet (about 0.1g of Astragalus polysaccharides) in river crab bait in a mortar, adding 100ml of water, ultrasonic treating for 10min, filtering, and measuring absorbance A2 at 490 nm. The 2h release of astragalus polysaccharide was calculated as follows:
the results show that the three release media release 2h, the scutellaria polysaccharide river crab bait coated pellet is complete, the cracking phenomenon does not occur, and the astragalus polysaccharide release degree is lower than 5% in 2h.
The invention carries out a series of immune challenge experimental researches. Research results show that the scutellaria polysaccharide can obviously inhibit the proliferation of aeromonas hydrophila; after the aeromonas hydrophila infects the eriocheir sinensis, the scutellaria baicalensis polysaccharide can improve the survival rate of the eriocheir sinensis; in the aspect of cellular immunity, the baical skullcap root polysaccharide can obviously improve the number and phagocytosis activity of the eriocheir sinensis hemolymph cells and inhibit apoptosis of the hemolymph cells; in the aspect of humoral immunity, the scutellaria polysaccharide can activate a TGF-beta signal pathway of the Eriocheir sinensis, inhibit the expression of inflammatory cytokines and simultaneously activate immune cells of an organism to generate the expression of a large number of genes such as lysozyme, peroxidase, antibacterial peptide and the like. This indicates that the scutellaria polysaccharide is an excellent immunopotentiator in regulating the immune response of eriocheir sinensis.
The invention has the beneficial effects that: antibacterial activity, can effectively inhibit the proliferation of aeromonas hydrophila; the individual level improves the survival rate of the eriocheir sinensis; in the aspect of innate immunity, the expression level of immune effector molecules can be obviously improved, and the expression of inflammatory cytokines can be inhibited, so that the immune enhancement function of the Eriocheir sinensis can be realized. The natural immunopotentiator for the eriocheir sinensis to resist the invasion of pathogens such as aeromonas hydrophila and the like has the characteristics of antibacterium, antivirus, no toxic or side effect, low pollution, low cost and simple preparation method, reduces the pollution in the baicalin extraction process, improves the additional value of the baicalin extraction industry, and meets the national requirements of energy conservation, emission reduction and carbon neutralization and carbon peak reaching.
Drawings
FIG. 1 is a graph showing the effect of Scutellaria baicalensis Georgi polysaccharides on the total number of Eriocheir sinensis hemolymphocytes;
FIG. 2 is a graph showing the effect of the Scutellariae radix polysaccharide on phagocytic activity of Eriocheir sinensis hemolymph cells;
FIG. 3 is a graph showing the effect of Scutellaria baicalensis Georgi polysaccharides on phagocytic index of Eriocheir sinensis blood lymphocytes.
Detailed Description
The following detailed description will describe specific embodiments of the present invention.
Example 1
Bacteriostatic experiments: aeromonas hydrophila (provided by Song Lin university of Dalian sea university) is provided,
making filter paper into circular paper sheet with diameter of 6mm with a puncher, sterilizing at high temperature, soaking Scutellariae radix polysaccharide water extractive solution, drying, spreading the paper sheet on a uniform plate coated with bacteria, and soaking the paper sheet with sterile distilled water as contrast. After marking, the mixture is placed upside down in a constant temperature illumination incubator at 37 ℃ for culture, and the size of the inhibition zone is observed and recorded after 24 hours (Table 1).
TABLE 1 zone of inhibition of Aeromonas hydrophila by Scutellaria polysaccharides
Toxicity attack experiment
The eriocheir sinensis is divided into Blank group, PBS group, aeromonas hydrophila treatment group (Ah group) and scutellaria polysaccharide treatment group (the concentrations of scutellaria polysaccharide are respectively 0.5%, 1.0% and 2.0%, and are respectively marked as HQ0.5, HQ1.0 and HQ 2.0), and each group contains 50 river crabs. Wherein, the group HQ0.5, HQ1.0 and HQ2.0 are injected with baical skullcap root polysaccharide with different concentrations 24 hours in advance, after 24 hours, the group Ah, HQ0.5, HQ1.0 and HQ2.0 are respectively injected with aeromonas hydrophila to be treated, and after the injection, 6 river crabs are randomly sampled from each experimental group at 0h, 3h, 6h, 12h, 24h and 48h respectively.
Mortality test
According to the requirements of the challenge experiment, the death rate of the eriocheir sinensis in each experimental group is measured at 0h, 3h, 6h, 12h, 24h and 48h respectively (Table 2). After 48 hours of infection of aeromonas hydrophila, the death rate of river crabs is 0 in Blank group and PBS group, the death rate of river crabs is 76% in Ah group, and the death rates of HQ0.5, HQ1.0 and HQ2.0 are 32%, 26% and 18% respectively along with the increase of the concentration of the scutellaria baicalensis polysaccharide, which shows that the survival rate of the eriocheir sinensis can be obviously improved by the scutellaria baicalensis polysaccharide after the aeromonas hydrophila is infected.
TABLE 2 Effect of different concentrations of Scutellaria baicalensis polysaccharides on Eriocheir sinensis mortality following Aeromonas hydrophila infection
Note: 50 Eriocheir sinensis were selected from each experimental group.
Scutellaria baicalensis polysaccharide for regulating humoral immunity of Eriocheir sinensis
According to the requirements of the challenge experiment, the change of the immune index in the Eriocheir sinensis Henlingba is analyzed by using a Nanjing-constructed lysozyme, phenol oxidase and TNF-alpha detection kit.
Experimental results show that the lysozyme activity, the phenol oxide enzyme activity and the TNF-alpha concentration of the Eriocheir sinensis in the haemolymph are not obviously changed in the Blank group and the PBS group within 0-48 h after the aeromonas hydrophila is attacked.
Lysozyme: after the river crabs are treated by aeromonas hydrophila for 6 hours, 12 hours and 24 hours, the activities of lysozyme are respectively improved by 56.72 percent, 52.18 percent and 42.53 percent; after the HQ0.5 group of river crabs are treated by aeromonas hydrophila for 6 hours and 12 hours, the activities of lysozyme are respectively improved by 66.34 percent and 58.77 percent; after the HQ1.0 group of river crabs are treated by aeromonas hydrophila for 6 hours and 12 hours, the activities of lysozyme are respectively improved by 76.54 percent and 83.48 percent; after the HQ2.0 group of river crabs are treated by aeromonas hydrophila for 6h and 12h, the activities of lysozyme are respectively improved by 89.87 percent and 95.46 percent (Table 3).
TABLE 3 Effect of Scutellaria baicalensis polysaccharide on Eriocheir sinensis hemolymph lysozyme activity after Aeromonas hydrophila infection
Phenol oxidizing enzyme: after the river crabs are processed by Aeromonas hydrophila for 3h, 6h and 12h, the activity of the phenol oxidase is respectively improved by 60.99 percent, 52.99 percent and 43.46 percent; the activity of the lysozyme of HQ0.5 group river crabs is respectively improved by 84.12 percent, 69.38 percent and 52.86 percent; the activity of the lysozyme of HQ1.0 group of river crabs is respectively improved by 91.19 percent, 56.67 percent and 51.46 percent; the activity of the lysozyme of HQ2.0 group of river crabs is respectively improved by 106.99 percent, 81.32 percent and 57.31 percent; (Table 4).
TABLE 4 Effect of Scutellaria baicalensis Georgi polysaccharide on Eriocheir sinensis hemolymph phenol oxidase Activity after Aeromonas hydrophila infection
TNF-alpha concentration determination: after the aeromonas hydrophila is treated, the concentration of an inflammatory cytokine TNF-alpha in the river crab blood lymph of the HQ0.5 group, the HQ01.0 group and the HQ2.0 group is obviously lower than that of the river crab of the Ah group, which indicates that the aeromonas hydrophila can initiate the inflammatory reaction of the river crab, and the scutellaria polysaccharide can obviously inhibit the inflammatory reaction of the river crab initiated by the aeromonas hydrophila (Table 5).
TABLE 5 Change in the concentration of haemolymph TNF-alpha from Eriocheir sinensis by Scutellaria baicalensis polysaccharides following Aeromonas hydrophila infection
Scutellaria baicalensis polysaccharide for regulating immunity of Eriocheir sinensis cells
According to the requirements of the toxicity attacking experiment, the ratio of hemolymph and anticoagulant is 1:1, extracting 500ml of hemolymph from the base of the chelicea sinensis, quickly putting into a 1.5ml centrifuge tube, centrifuging at 4 ℃,1000rpm for 3min, removing supernatant, adding 100ul 4% paraformaldehyde, slightly blowing to disperse cells, and fixing at room temperature for 15min.
The fixed hemolymphocytes are dropped into the counting interval of a blood counting chamber, a cover glass is covered, the total number of all types of hemocytes is counted under a microscope, and each sample 2 is repeated 5 times (figure 1). Experimental results show that the total number of hemolymph cells of the Eriocheir sinensis is obviously reduced by the infection of the Aeromonas hydrophila, and the quantity of the hemolymph cells in the river crab infected by the Aeromonas hydrophila can be obviously increased by the Scutellaria polysaccharide, so that the apoptosis of the hemolymph cells caused by the Aeromonas hydrophila is inhibited, and the cellular immunity of the river crab is provided.
The Aeromonas hydrophila in logarithmic culture phase is subjected to heat inactivation treatment (85 ℃,30 min), then is centrifuged, and is added with NaHCO containing 1 mg/mL Fluorescein Isothiocyanate (FITC) 3 (0.1 mol/L, pH 9.0), resuspension, incubation at 25 ℃ for 3h, and PBS washing of the bacteria to remove excess FITC. The labeled bacteria were resuspended in PBS and adjusted to a concentration of 1X 10 8 CFU/mL, stored at-20 ℃ until use. Hemolymphocytes were mixed with FITC-labeled aeromonas hydrophila at a ratio of 1. 200 mu L of the mixture is placed on glass slides, and is placed in a wet box for sedimentation for 30min, and then is washed with sterilized normal saline for 2-3 times, and then trypan blue is added to quench the extracellular FITC, and after 10min, is washed with sterilized normal saline for 2-3 times, after drying at room temperature, glycerol is added for sealing, and the phagocytosis activity of the hemolymph cells is observed under a fluorescence microscope, and about 100 blood cells are counted on each glass slide. Phagocytic activity includesPhagocytic Rate (PR) and phagocytic index (PL), where PR = number of phagocytic cells/total number of cells x 100%; PL = total number of bacteria in phagocytic cells/total number of phagocytic cells.
Experimental results show that after the aeromonas hydrophila is infected, the phagocytosis activity of the eriocheir sinensis hemolymph cells is remarkably improved within 6-24 hours, in the time period, the baicalein can further improve the phagocytosis activity of the hemolymph cells, and in the baicalein treatment group, the phagocytosis rate of the eriocheir sinensis hemolymph cells is remarkably higher than that of the eriocheir sinensis hemolymph cells in the Ah group at the same time point (figure 2).
The result of phagocytosis indexes of the hemolymph cells shows that the phagocytosis indexes of the hemolymph cells of the Eriocheir sinensis are obviously improved within 6-12 hours after the Aeromonas hydrophila is infected, the phagocytosis rate of the hemolymph cells can be further improved by the baicalein in the period of time, and the phagocytosis rate of the hemolymph cells of the Eriocheir sinensis is obviously higher than that of the hemolymph cells of the Eriocheir sinensis in an Ah group at the same time point in a baicalein treatment group (figure 3).
Claims (8)
1. An application of an immunopotentiator based on scutellaria baicalensis polysaccharide in Eriocheir sinensis cultivation is characterized by comprising the following steps of: adding hydrochloric acid into the scutellaria baicalensis extracting solution for precipitation, filtering to obtain filtrate (pH = 1.0-2.0), and adjusting the pH value to 6.0-7.0; concentrating at low temperature (50-70 deg.C) and vacuum degree (0.4-0.8 Mpa) to obtain concentrated solution.
2. The use of the scutellaria polysaccharide-based immunopotentiator according to claim 1 in eriocheir sinensis cultivation, wherein the immunopotentiator is characterized in that: alcohol precipitation of scutellaria polysaccharide: adding edible alcohol (alcohol content greater than 90% (v/v)) into the concentrated solution until the alcohol content reaches about 70% (v/v), precipitating at low temperature for 3-8 hr, and filtering to obtain Scutellariae radix polysaccharide precipitate; filter-pressing the precipitate to obtain a solid, then slicing (slice thickness: 0.5 mm-2.0 mm), placing the polysaccharide slices in an oven at 40-60 ℃ for 0.5-3.0 h, and obtaining the dried scutellaria baicalensis polysaccharide.
3. The use of the scutellaria polysaccharide-based immunopotentiator according to claim 1 in eriocheir sinensis cultivation, wherein the immunopotentiator is characterized in that: the volume of the baicalin concentrated solution is 10-30% of the volume of the baicalin filtrate; the vacuum degree of the concentration tank is adjusted at any time according to the concentration temperature.
4. The use of the scutellaria polysaccharide-based immunopotentiator according to claim 2 in eriocheir sinensis cultivation, wherein: the density and hardness of the filter-pressed scutellaria polysaccharide meet the slicing requirements, and the thickness of the slices is adjusted according to the density and hardness; and adjusting the temperature and time parameters of the dryer according to the slice thickness.
5. The use of the scutellaria polysaccharide-based immunopotentiator according to claim 2 for Eriocheir sinensis cultivation, wherein the immunopotentiator comprises: preparing scutellaria polysaccharide bait: adding the dried scutellaria polysaccharide into the river crab basic feed, including fish meal, river snail meat powder, peanut cake powder, rice flour starch, etc., wherein the content percentage of the scutellaria polysaccharide is 1.0% -3.0%.
6. The use of the scutellaria polysaccharide-based immunopotentiator according to claim 5 in Eriocheir sinensis cultivation, wherein the immunopotentiator comprises: the stability of the scutellaria polysaccharide bait is as follows: adjusting the content proportion of the scutellaria polysaccharide and other components in the river crab bait, and controlling the water content of the raw material to be 10-15% and the crude fiber component to be 3.0-6.0%; the stability of the bait in water is more than 2h.
7. The use of the scutellaria polysaccharide-based immunopotentiator according to claim 6 for Eriocheir sinensis cultivation, wherein the immunopotentiator comprises: feeding river crab with the processed river crab bait, analyzing the effect of the content of the scutellaria baicalensis polysaccharide in the bait on the immunity enhancement of the river crab, and finally determining the optimal proportion of the scutellaria baicalensis polysaccharide in the bait to be about 2.0%.
8. Use of the scutellaria polysaccharide-based immunopotentiator according to claims 1-7 in eriocheir sinensis cultivation, wherein: the baicalein derived from the baicalin extraction waste liquid can be used as an immunopotentiator to be added into Eriocheir sinensis bait for regulating the immunity of Eriocheir sinensis.
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