CN116831069A - Method for improving transportation survival rate of pelteobagrus fulvidraco - Google Patents
Method for improving transportation survival rate of pelteobagrus fulvidraco Download PDFInfo
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- CN116831069A CN116831069A CN202310782325.9A CN202310782325A CN116831069A CN 116831069 A CN116831069 A CN 116831069A CN 202310782325 A CN202310782325 A CN 202310782325A CN 116831069 A CN116831069 A CN 116831069A
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- polysaccharide
- pelteobagrus fulvidraco
- water
- composite
- fish
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
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-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
- A01K61/13—Prevention or treatment of fish diseases
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
- A01K63/02—Receptacles specially adapted for transporting live fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/10—Organic substances
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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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Abstract
The invention discloses a method for improving the transportation survival rate of pelteobagrus fulvidraco, which comprises the following steps: (1) Selecting healthy pelteobagrus fulvidraco juvenile fish, putting the juvenile pelteobagrus fulvidraco into a culture water vat, injecting water into the water vat, adding composite polysaccharide nanoparticles and composite probiotics, and fasted temporarily raising the pelteobagrus fulvidraco juvenile fish in the vat for 1-3 days; (2) Filling water and the juvenile pelteobagrus fulvidraco fasted and temporarily raised in the step (1) into an oxygen bag special for aquatic products, immediately oxygenating and sealing the bags, ensuring the oxygen content to be 6-7mg/L, and placing the bags into a carton with foam for transportation; the compound polysaccharide nanoparticle comprises chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide. The composite polysaccharide nanoparticle is beneficial to the absorption of polysaccharide and acts on tissues, increases the solubility and dispersibility of water and the absorption of fish bodies to the polysaccharide nanoparticle, effectively avoids side effects caused by the pollution of water due to the accumulation of polysaccharide, reduces the time and economic cost caused by the addition of the polysaccharide to feed, and further realizes the alleviation of fish stress under transportation stress.
Description
Technical Field
The invention belongs to the technical field of fish transportation, and particularly relates to a method for improving the transportation survival rate of pelteobagrus fulvidraco.
Background
Aquaculture is a rapidly growing industry worldwide providing one of the most sustainable forms of dietary protein and nutrition production. According to a report of the grain and agriculture organization 2020, the fish provides more than 33 hundred million people with 20% of their per-human animal protein intake worldwide, and the world aquaculture fish yield reaches 8210 ten thousand tons in 2018. With the development of economy, demands for live water products, particularly high-value varieties, are increasing, and fish transportation becomes a key to live fish consumption. Transportation involves capturing, loading, transporting, unloading and stocking, and many studies have demonstrated that aquatic animals are severely stressed during transportation, negatively affecting the aquaculture industry and economically losing. These stresses lead to physiological reactions such as the release of catecholamines and corticosteroids, and to increased blood glucose levels, or reduced immune system function, leading to disease and death. In addition, the continuous stress during the transportation of fish will lead to quality degradation.
Yellow catfish is also called Huang Jia fish, yellow bone fish, yellow thorn fish and Long Dun fish. Distributed in the river basin such as Yangtze river, yellow river, zhujiang river and Heilongjiang river in China, the Chinese catfish has sweet and flat nature, can benefit spleen and stomach, induce diuresis to alleviate edema, has tender meat quality, rich nutrition and extremely high economic value, and because the pectoral fin and dorsal fin of yellow catfish grow with hard spines, the skin of the fish body is easily scratched during transportation of live fish, secondary bacterial infection and mold infection are caused, and the survival rate and quality of yellow catfish are affected to a certain extent.
Disclosure of Invention
The invention aims to overcome the technical defects, and provides a method for improving the survival rate of pelteobagrus fulvidraco, which solves the technical problems that skin of the pelteobagrus fulvidraco is easy to scratch during transportation of live pelteobagrus fulvidraco, secondary bacterial infection and mould infection are caused, the survival rate and quality of pelteobagrus fulvidraco are affected to a certain extent, and the like in the prior art.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
a method for improving the transportation survival rate of pelteobagrus fulvidraco comprises the following specific steps:
(1) Selecting healthy pelteobagrus fulvidraco juvenile fish, putting the juvenile pelteobagrus fulvidraco into a culture water vat, injecting water into the water vat, adding composite polysaccharide nanoparticles and composite probiotics, and fasted temporarily raising the pelteobagrus fulvidraco juvenile fish in the vat for 1-3 days;
(2) Filling water and the juvenile pelteobagrus fulvidraco fasted and temporarily raised in the step (1) into an oxygen bag special for aquatic products, immediately oxygenating and sealing the bags, ensuring the oxygen content to be 6-7mg/L, and placing the bags into a carton with foam for transportation;
the compound polysaccharide nanoparticle comprises chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide.
Preferably, the preparation method of the composite polysaccharide nanoparticle comprises the following steps: respectively weighing chitosan, astragalus polysaccharide, xylooligosaccharide, roxburgh rose polysaccharide and matrimony vine polysaccharide, adding deionized water, stirring at 20-30deg.C for 40-70min to dissolve completely, and preparing into 5 polysaccharide solutions; then, under the condition of ultrasonic dispersion, the 5 polysaccharide solutions are sequentially and uniformly dripped into an ethanol solution, the mixture is stirred for 80-120min at 25-30 ℃, then the mixture is subjected to rotary evaporation concentration at 40-60 ℃, when the solvent is evaporated to 2/3 of the original volume, deionized water is added to complement the original volume, the rotary evaporation concentration and the deionized water supplementing operation are repeated for 5 times, and then the mixed polysaccharide solution is subjected to freeze drying for 24-32h, so that the composite polysaccharide nanoparticle is obtained.
Preferably, the weight of the juvenile pelteobagrus fulvidraco in the step (1) is 7.77+/-1.70 g, the height of a culture water tank is 78cm, the diameter of the culture water tank is 100cm, the temperature of the culture water is 20-25 ℃, and the dissolved oxygen is 6-7mg/L, ph 7.8.8-8.4.
Preferably, the concentration of the composite polysaccharide nanoparticle in the step (1) is 0.02-0.3g/L, the composite probiotics are lactobacillus and bacillus subtilis, and the concentration of the composite probiotics is 0.1-0.3g/L.
Preferably, the mass ratio of the chitosan to the astragalus polysaccharide to the xylooligosaccharide to the rosa roxburghii polysaccharide to the lycium barbarum polysaccharide is 1-12:3-23:1.5-6:1-8:8-30 parts; the concentration of polysaccharide solutions prepared from chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide are respectively as follows: 10-120mg/L, 30-230mg/L, 15-60mg/L, 10-80mg/L and 80-300mg/L.
Preferably, the ultrasonic dispersing time is 10-30min and the power is 40-60W.
Preferably, the transport temperature is 20-25 ℃.
Compared with the prior art, the invention has the following beneficial effects:
according to the invention, the composite polysaccharide nanoparticles are added into temporary culture water, and the nano drug-carrying system can be uniformly dispersed after being dissolved in water due to small particle size, large specific surface area and high solubility, so that the absorption of polysaccharide and the action on tissues are facilitated, the water solubility and dispersibility are increased, the absorption of fish bodies on the water are improved, the side effect caused by the pollution of water due to the accumulation of polysaccharide is effectively avoided, the time and economic cost caused by the addition into feed are reduced, the relief of fish stress under transportation stress is realized, and the survival rate of pelteobagrus fulvidraco in the transportation process is remarkably improved.
The traditional Chinese medicine polysaccharide has various biological activities and no toxic or side effect, and is a green, safe and residue-free fish feed additive with great development potential. The polysaccharide can promote the production of more than 50 cytokines such as interferon, tumor necrosis factor, interleukin, etc. Plant polysaccharides can also achieve an increase in immunity by increasing the level of immunoglobulins in the body. Different levels of wolfberry polysaccharide can obviously promote T, B lymphocyte proliferation, astragalus polysaccharide can also promote T lymphocyte proliferation, animal organism cellular immunity is enhanced, rosa roxburghii polysaccharide can improve the immunity of fish, and a proper amount of compound polysaccharide can be a safe and effective immunostimulant, so that the humoral immune response of fish can be obviously enhanced. Xylo-oligosaccharide has prebiotic characteristics in aquaculture, and prebiotics have higher stability in an acidic environment of animal intestinal tracts, can promote the sugar metabolism level in fish bodies while improving the immunity of organisms, and are widely used in aquaculture industry. The combination of the prebiotics and the Chinese herbal medicine polysaccharide can improve the active ingredients and biological titers of the Chinese herbal medicine polysaccharide, and fully exert the synergistic effect between the two. The combination of the traditional Chinese medicine polysaccharide and the prebiotics can improve the oxidation resistance and the immunity of fish more than the single use of the prebiotics.
The compound polysaccharide has advantages in regulating and controlling 4 kinds of fish intestinal probiotics metabolites relative to single polysaccharide, and realizing functional complementation of multiple polysaccharides. Each polysaccharide has respective advantages in promoting the growth and acid production of various microorganisms, wherein the lycium barbarum polysaccharide has stronger probiotic activity on lactobacillus and bifidobacterium, and can promote the accumulation of propionic acid and isovaleric acid; chitosan has weaker proliferation promoting activity on mixed bacteria, but has optimal life-benefiting property on FP bacteria. The astragalus polysaccharide, the medlar polysaccharide and other polysaccharides can promote the growth of different intestinal probiotics, have the characteristic of functional complementation, and the compound polysaccharide obtained by scientifically combining the polysaccharides can simultaneously exert the advantages of the polysaccharides, has better proliferation promoting activity on all probiotics, particularly the bacteroides, achieves the aim of synergy, and further maintains the physiological activity of fish.
Meanwhile, compared with single polysaccharide, the activity and the effect of the compound polysaccharide are more obvious, and the compound polysaccharide can play a role in antioxidation and protection by improving the enzyme activity of a defense system, reducing the accumulation of free radicals in the body, reducing the damage of the free radicals to cells and other ways. The single polysaccharide has certain probiotics activity, but the immunity enhancing activity of the single polysaccharide is weaker, and only a few bacteria in several intestinal probiotics can hydrolyze and utilize the polysaccharide, so that when the single polysaccharide is applied to the intestinal tracts of fish bodies, only a few bacteria take the growth advantage. And the compound polysaccharide prepared by mixing a plurality of polysaccharides in a specific proportion can play a role of synergy. The more complex polysaccharide combinations are more readily available to more microorganisms and thus exert better effects in maintaining fish intestinal homeostasis, regulating intestinal flora metabolism. Compared with single polysaccharide, the compound polysaccharide has more advantages in promoting microorganisms to generate active peptide, can accumulate more dipeptides with ACE inhibitory activity, DPP-IV inhibitory activity and alpha-glucosidase inhibitory activity, and has greater hypoglycemic or antioxidant potential.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
It is emphasized in particular, unless otherwise specified, that the materials or reagents of the invention are commercially available.
Example 1
A method for improving the transportation survival rate of pelteobagrus fulvidraco comprises the following specific steps:
(1) Selecting healthy pelteobagrus fulvidraco juvenile fish, putting the juvenile pelteobagrus fulvidraco into a culture water vat, injecting water into the water vat, adding composite polysaccharide nanoparticles and composite probiotics, and fasted temporarily raising the pelteobagrus fulvidraco juvenile fish in the vat for 1 day;
(2) Filling water and young pelteobagrus fulvidraco fasted and temporarily raised in the step (1) into an oxygen bag special for aquatic products, immediately oxygenating and sealing the bags, ensuring the oxygen content to be 6mg/L, putting the bags into a paper box with foam inside for transportation, wherein the transportation density is 1:4 of fish-water ratio (m fish: m water), the transportation time is 8 hours, and the transportation temperature is 25 ℃;
the compound polysaccharide nanoparticle comprises chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide.
Further, the preparation method of the composite polysaccharide nanoparticle comprises the following steps: respectively weighing chitosan, astragalus polysaccharide, xylooligosaccharide, roxburgh rose polysaccharide and matrimony vine polysaccharide, adding deionized water, stirring at 20deg.C for 40min to fully dissolve, and preparing 5 polysaccharide solutions; then, under the condition of ultrasonic dispersion, the 5 polysaccharide solutions are sequentially and uniformly dripped into an ethanol solution, the mixture is stirred for 80min at 25 ℃, then the mixture is subjected to rotary evaporation concentration at 40 ℃, when the solvent is evaporated to 2/3 of the original volume, deionized water is added to complement the original volume, the rotary evaporation concentration and the deionized water supplementing operation are repeated for 5 times, and then the mixed polysaccharide solution is subjected to freeze drying for 24h, so that the composite polysaccharide nanoparticle is obtained.
Further, in the step (1), the weight of the juvenile pelteobagrus fulvidraco is 6.07g, the height of a culture water vat is 78cm, the diameter of the culture water vat is 100cm, the temperature of the culture water is 20 ℃, and the dissolved oxygen is 6mg/L, ph and is 7.8.
Further, the adding concentration of the composite polysaccharide nanoparticle in the step (1) is 0.02g/L, the composite probiotics are lactobacillus and bacillus subtilis, and the adding concentration of the composite probiotics is 0.1g/L.
Further, the concentrations of polysaccharide solutions prepared from chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide are respectively as follows: 10mg/L, 30mg/L, 15mg/L, 10mg/L and 80mg/L.
Further, the ultrasonic dispersing time is 10min, and the power is 40W.
Further, the transport temperature is 20 ℃.
Example 2
A method for improving the transportation survival rate of pelteobagrus fulvidraco comprises the following specific steps:
(1) Selecting healthy pelteobagrus fulvidraco juvenile fish, putting the juvenile pelteobagrus fulvidraco into a culture water vat, injecting water into the water vat, adding composite polysaccharide nanoparticles and composite probiotics, and fasted temporarily raising the pelteobagrus fulvidraco juvenile fish in the vat for 2 days;
(2) Filling water and young pelteobagrus fulvidraco fasted and temporarily raised in the step (1) into an oxygen bag special for aquatic products, immediately oxygenating and sealing the bags, ensuring the oxygen content to be 6.5mg/L, putting the bags into a paper box with foam inside for transportation, wherein the transportation density is 1:4 (m fish: m water), the transportation time is 8 hours, and the transportation temperature is 25 ℃;
the compound polysaccharide nanoparticle comprises chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide.
Further, the preparation method of the composite polysaccharide nanoparticle comprises the following steps: respectively weighing chitosan, astragalus polysaccharide, xylooligosaccharide, roxburgh rose polysaccharide and matrimony vine polysaccharide, adding deionized water, stirring at 25deg.C for 55min to fully dissolve, and preparing 5 polysaccharide solutions; then, under the condition of ultrasonic dispersion, the 5 polysaccharide solutions are sequentially and uniformly dripped into an ethanol solution, stirred at 27 ℃ for 100min, concentrated by rotary evaporation at 50 ℃, when the solvent is evaporated to 2/3 of the original volume, deionized water is added to complement the original volume, the rotary evaporation concentration and the deionized water supplementing operation are repeated for 5 times, and then the mixed polysaccharide solution is freeze-dried for 28h, so that the composite polysaccharide nanoparticle is obtained.
Further, in the step (1), the weight of the juvenile pelteobagrus fulvidraco is 7.77g, the height of a culture water vat is 78cm, the diameter of the culture water vat is 100cm, the temperature of the culture water is 23 ℃, and the dissolved oxygen is 6.5mg/L, ph and is 8.7.
Further, the adding concentration of the composite polysaccharide nanoparticle in the step (1) is 0.17g/L, the composite probiotics are lactobacillus and bacillus subtilis, and the adding concentration of the composite probiotics is 0.2g/L.
The concentration of polysaccharide solutions prepared from chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide are respectively as follows: 65mg/L, 130mg/L, 37.5mg/L, 45/L and 190mg/L.
Further, the ultrasonic dispersing time is 20min, and the power is 50W.
Further, the transport temperature was 23 ℃.
Example 3
A method for improving the transportation survival rate of pelteobagrus fulvidraco comprises the following specific steps:
(1) Selecting healthy pelteobagrus fulvidraco juvenile fish, putting the juvenile pelteobagrus fulvidraco into a culture water vat, injecting water into the water vat, adding composite polysaccharide nanoparticles and composite probiotics, and fasted temporarily raising the pelteobagrus fulvidraco juvenile fish in the vat for 3 days;
(2) Filling water and young pelteobagrus fulvidraco fasted and temporarily raised in the step (1) into an oxygen bag special for aquatic products, immediately oxygenating and sealing the bags, ensuring the oxygen content to be 7mg/L, putting the bags into a paper box with foam inside for transportation, wherein the transportation density is 1:4 of fish-water ratio (m fish: m water), the transportation time is 8 hours, and the transportation temperature is 25 ℃;
the compound polysaccharide nanoparticle comprises chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide.
Further, the preparation method of the composite polysaccharide nanoparticle comprises the following steps: respectively weighing chitosan, astragalus polysaccharide, xylooligosaccharide, roxburgh rose polysaccharide and matrimony vine polysaccharide, adding deionized water, stirring at 30deg.C for 70min to fully dissolve, and preparing 5 polysaccharide solutions; then, under the condition of ultrasonic dispersion, the 5 polysaccharide solutions are sequentially and uniformly dripped into an ethanol solution, the mixture is stirred for 120min at 30 ℃, then the mixture is subjected to rotary evaporation concentration at 60 ℃, when the solvent is evaporated to 2/3 of the original volume, deionized water is added to complement the original volume, the rotary evaporation concentration and the deionized water supplementing operation are repeated for 5 times, and then the mixed polysaccharide solution is subjected to freeze drying for 32h, so that the composite polysaccharide nanoparticle is obtained.
Further, in the step (1), the weight of the juvenile pelteobagrus fulvidraco is 9.47g, the height of a culture water vat is 78cm, the diameter of the culture water vat is 100cm, the temperature of the culture water is 25 ℃, and the dissolved oxygen is 7mg/L, ph 8.4.4.
Further, the adding concentration of the composite polysaccharide nanoparticle in the step (1) is 0.3g/L, the composite probiotics are lactobacillus and bacillus subtilis, and the adding concentration of the composite probiotics is 0.3g/L.
The concentration of polysaccharide solutions prepared from chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide are respectively as follows: 120mg/L, 230mg/L, 60mg/L, 80mg/L and 300mg/L.
Further, the ultrasonic dispersing time is 30min, and the power is 60W.
Further, the transport temperature is 25 ℃.
Comparative example 1
A method for improving the transportation survival rate of pelteobagrus fulvidraco comprises the following specific steps:
(1) Selecting healthy pelteobagrus fulvidraco juvenile fish, putting the juvenile pelteobagrus fulvidraco into a culture water vat, injecting water into the water vat, adding composite polysaccharide nanoparticles and composite probiotics, and fasted temporarily raising the pelteobagrus fulvidraco juvenile fish in the vat for 1 day;
(2) Filling water and young pelteobagrus fulvidraco fasted and temporarily raised in the step (1) into an oxygen bag special for aquatic products, immediately oxygenating and sealing the bags, ensuring the oxygen content to be 6mg/L, putting the bags into a paper box with foam inside for transportation, wherein the transportation density is 1:4 of fish-water ratio (m fish: m water), the transportation time is 8 hours, and the transportation temperature is 25 ℃;
the compound polysaccharide nanoparticle comprises chitosan and astragalus polysaccharide.
Further, the preparation method of the composite polysaccharide nanoparticle comprises the following steps: respectively weighing chitosan and astragalus polysaccharide, adding deionized water, stirring at 20deg.C for 40min to dissolve completely, and preparing into 2 polysaccharide solutions; then, under the condition of ultrasonic dispersion, the 2 polysaccharide solutions are sequentially and uniformly dripped into an ethanol solution, the mixture is stirred at 25 ℃ for 80min, then the mixture is subjected to rotary evaporation concentration at 40 ℃, when the solvent is evaporated to 2/3 of the original volume, deionized water is added to complement the original volume, the rotary evaporation concentration and the deionized water supplementing operation are repeated for 5 times, and then the mixed polysaccharide solution is subjected to freeze drying for 24h, so that the composite polysaccharide nanoparticle is obtained.
Further, in the step (1), the weight of the juvenile pelteobagrus fulvidraco is 6.07g, the height of a culture water vat is 78cm, the diameter of the culture water vat is 100cm, the temperature of the culture water is 20 ℃, and the dissolved oxygen is 6mg/L, ph and is 7.8.
Further, the adding concentration of the composite polysaccharide nanoparticle in the step (1) is 0.02g/L, the composite probiotics are lactobacillus and bacillus subtilis, and the adding concentration of the composite probiotics is 0.1g/L.
Further, the concentration of the polysaccharide solution prepared from the chitosan and the astragalus polysaccharide is respectively as follows: 10mg/L, 30mg/L.
Further, the ultrasonic dispersing time is 10min, and the power is 40W.
Further, the transport temperature is 20 ℃.
The difference between this comparative example and example 1 is that the comparative example did not incorporate xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide.
Comparative example 2
A method for improving the transportation survival rate of pelteobagrus fulvidraco comprises the following specific steps:
(1) Selecting healthy pelteobagrus fulvidraco juvenile fish, putting the juvenile pelteobagrus fulvidraco into a culture water vat, injecting water into the water vat, adding composite polysaccharide nanoparticles and composite probiotics, and fasted temporarily raising the pelteobagrus fulvidraco juvenile fish in the vat for 1 day;
(2) Filling water and young pelteobagrus fulvidraco fasted and temporarily raised in the step (1) into an oxygen bag special for aquatic products, immediately oxygenating and sealing the bags, ensuring the oxygen content to be 6mg/L, putting the bags into a paper box with foam inside for transportation, wherein the transportation density is 1:4 of fish-water ratio (m fish: m water), the transportation time is 8 hours, and the transportation temperature is 25 ℃;
the compound polysaccharide nanoparticle comprises astragalus polysaccharide and xylooligosaccharide.
Further, the preparation method of the composite polysaccharide nanoparticle comprises the following steps: respectively weighing astragalus polysaccharide and xylooligosaccharide, adding deionized water, stirring at 20deg.C for 40min to dissolve completely, and preparing into 2 polysaccharide solutions; then, under the condition of ultrasonic dispersion, the 2 polysaccharide solutions are sequentially and uniformly dripped into an ethanol solution, the mixture is stirred at 25 ℃ for 80min, then the mixture is subjected to rotary evaporation concentration at 40 ℃, when the solvent is evaporated to 2/3 of the original volume, deionized water is added to complement the original volume, the rotary evaporation concentration and the deionized water supplementing operation are repeated for 5 times, and then the mixed polysaccharide solution is subjected to freeze drying for 24h, so that the composite polysaccharide nanoparticle is obtained.
Further, in the step (1), the weight of the juvenile pelteobagrus fulvidraco is 6.07g, the height of a culture water vat is 78cm, the diameter of the culture water vat is 100cm, the temperature of the culture water is 20 ℃, and the dissolved oxygen is 6mg/L, ph and is 7.8.
Further, the adding concentration of the composite polysaccharide nanoparticle in the step (1) is 0.02g/L, the composite probiotics are lactobacillus and bacillus subtilis, and the adding concentration of the composite probiotics is 0.1g/L.
Further, the concentration of the polysaccharide solution prepared from astragalus polysaccharide and xylooligosaccharide is respectively as follows: 30mg/L, 15mg/L.
Further, the ultrasonic dispersing time is 10min, and the power is 40W.
Further, the transport temperature is 20 ℃.
The difference between this comparative example and example 1 is that chitosan, rosa roxburghii polysaccharide and lycium barbarum polysaccharide are not introduced.
The juvenile pelteobagrus fulvidraco transported in examples 1-3 and comparative examples 1-2 were subjected to the following index determination:
the oxygen consumption of the organism is increased during transportation, the generation of Reactive Oxygen Species (ROS) is promoted, the organism is damaged, and the antioxidation defense plays a key role in the removal of the ROS. Superoxide dismutase (SOD) plays an important role in the reaction to O2-and Catalase (CAT) relieves oxidative stress by degrading hydrogen peroxide. The pelteobagrus fulvidraco fry of the temporary rearing group added with the composite additive is stressed less than that of the group without the composite additive. Similarly, the overall CAT level is increased after transportation, and the temporary rearing group has minimum change, which indicates that temporary rearing of various additives can relieve the pelteobagrus fulvidraco fry against transportation stress.
Malondialdehyde (MDA) is a product of peroxidation of body lipids, and the content of MDA can be used as an important index of peroxide accumulation and degree of oxidative stress. MDA has been found to cause damage to the protein structure, primarily because it reacts with some amino acids within itself; MDA also has a cellular damage, mainly in the sense of destroying the respiratory function in mitochondria and also the associated dehydrogenases. Thus, the severity of the attack of ROS and the destruction of cells can be indirectly reflected by the detection of MDA content. Temporary rearing of the two composite additives also plays a role in reducing MDA level, wherein the MDA level of astragalus polysaccharide and chitosan group is lower.
IgM is a specific immune factor which is generated by B lymphocyte differentiation, is combined with antigen specificity, and has the functions of inhibiting pathogenic activity and regulating antigen. Immunoglobulin M (IgM) is a specific immune factor in fish, but it does not directly exert its immune function, but rather acts to produce immunoglobulin when the fish is stimulated, so as to exert its immune function. The compound polysaccharide nanoparticle temporary rearing group promotes the IgM level in the pelteobagrus fulvidraco body, and can stimulate the fish body to play an immune function.
Lysozyme is one of the most important nonspecific immune factors, one of the key elements of innate immunity against bacterial invasion. Is also a critical humoral component of the innate immune system. The activity level of lysozyme in fish body is measured, and the state of nonspecific humoral immunity of fish can be reflected to a certain extent. Complement is an important component of the fish immune system, with C3 and C4 occupying important roles in the Complement system. The level of C3 not only can reflect the immune pathological damage condition of the organism, but also is an important physiological defending system of the organism, and has biological functions of sterilization and conditioning, immunoadsorption and the like after the activation. Complement is also a mediator that can link antibodies to phagocytes, can enhance humoral and specific immune functions, and can also serve an indispensable role in nonspecific immunity of organisms. The C3 and lysozyme levels in the transported pelteobagrus fulvidraco fries are increased. The levels of the compound polysaccharide nanoparticle temporary culture group C3 and lysozyme are higher than those of a control group without addition.
In fish, apoptosis can be induced by invasive bacteria, accompanied by activation of many intracellular proteases and endonucleases, and increased apoptosis compromises intestinal structural integrity. Depending on the involvement of caspases, the apoptotic pathway is divided into caspase-dependent and caspase-independent transduction pathways. Caspases involved in apoptosis include both initiating caspases (e.g., casp-2, casp-8, and Casp-9) and effector caspases (e.g., casp-3 and Casp-7), which play a major role in regulating apoptosis. The intestinal apoptosis level of the pelteobagrus fulvidraco fries after transportation is increased, and the transportation has adverse effects on the pelteobagrus fulvidraco fries. However, after the juvenile pelteobagrus fulvidraco with the compound polysaccharide nanoparticle temporarily-cultured group is transported, the apoptosis level of intestinal cells is lower than that of the juvenile pelteobagrus fulvidraco without the control group.
According to the invention, the composite polysaccharide nanoparticles are added into temporary culture water, and the nano drug-carrying system can be uniformly dispersed after being dissolved in water due to small particle size, large specific surface area and high solubility, so that the absorption of polysaccharide and the action on tissues are facilitated, the water solubility and dispersibility are increased, the absorption of fish bodies on the water are improved, the side effect caused by the pollution of water due to the accumulation of polysaccharide is effectively avoided, the time and economic cost caused by the addition into feed are reduced, the relief of fish stress under transportation stress is realized, and the survival rate of pelteobagrus fulvidraco in the transportation process is remarkably improved.
Transportation survival rate experiment:
1500 healthy pelteobagrus fulvidraco juvenile fish (weight of 7.77+/-1.70 g) are selected and put into a culture water vat (height of 78cm, diameter of 100 cm) at 20 ℃ with dissolved oxygen of 6-7mg/L, pG 7.8.8-8.4. Injecting 500L of water into four water tanks respectively, putting the fish into a fish tank according to the mass ratio of 1:4 (m fish: m water), respectively setting 4 groups (120 g polysaccharide nanoparticle group (example 1), 120g chitosan+astragalus polysaccharide (comparative example 1), 120g xylooligosaccharide+astragalus polysaccharide (comparative example 2) and no addition group), adding 50g composite probiotics (lactobacillus and bacillus subtilis), fasted and temporarily reared in the tanks for 3 days, filling the tanks into nylon oxygenation bags, sealing and filling pure oxygen, transporting in a water transportation mode, putting the transport bags into a simulated transportation incubator, simulating transportation vibration once every half hour, collecting samples after transportation for 0h, 4h, 8h, 12h, 16h and 24h, and recording survival rate.
Table 2 pelteobagrus fulvidraco fry transport survival rate
Table 2 records the survival rate of pelteobagrus fulvidraco fries before and after simulated transportation. After 20h of transportation, the survival rate after temporary culture of polysaccharide nanoparticle group is highest (100%), followed by chitosan+astragalus polysaccharide (55%). The survival rate of the xylooligosaccharide plus astragalus polysaccharide and the non-added group after 24 hours of transportation is 0.
The above description describes a preferred embodiment of the invention and should not be viewed as limiting the scope of the claims. Any modification, equivalent replacement and improvement without departing from the principle and spirit of the present invention should be considered as being within the scope of the claims of the present invention.
Claims (7)
1. A method for improving the transportation survival rate of pelteobagrus fulvidraco is characterized by comprising the following specific steps:
(1) Selecting healthy pelteobagrus fulvidraco juvenile fish, putting the juvenile pelteobagrus fulvidraco into a culture water vat, injecting water into the water vat, adding composite polysaccharide nanoparticles and composite probiotics, and fasted temporarily raising the pelteobagrus fulvidraco juvenile fish in the vat for 1-3 days;
(2) Filling water and the juvenile pelteobagrus fulvidraco fasted and temporarily raised in the step (1) into an oxygen bag special for aquatic products, immediately oxygenating and sealing the bags, ensuring the oxygen content to be 6-7mg/L, and placing the bags into a carton with foam for transportation;
the compound polysaccharide nanoparticle comprises chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide.
2. The method according to claim 1, wherein the preparation method of the composite polysaccharide nanoparticle comprises the following steps: respectively weighing chitosan, astragalus polysaccharide, xylooligosaccharide, roxburgh rose polysaccharide and matrimony vine polysaccharide, adding deionized water, stirring at 20-30deg.C for 40-70min to dissolve completely, and preparing into 5 polysaccharide solutions; then, under the condition of ultrasonic dispersion, the 5 polysaccharide solutions are sequentially and uniformly dripped into an ethanol solution, the mixture is stirred for 80-120min at 25-30 ℃, then the mixture is subjected to rotary evaporation concentration at 40-60 ℃, when the solvent is evaporated to 2/3 of the original volume, deionized water is added to complement the original volume, the rotary evaporation concentration and the deionized water supplementing operation are repeated for 5 times, and then the mixed polysaccharide solution is subjected to freeze drying for 24-32h, so that the composite polysaccharide nanoparticle is obtained.
3. The method according to claim 1, wherein the weight of the juvenile pelteobagrus fulvidraco in the step (1) is 7.77+/-1.70 g, the height of the culture water tank is 78cm, the diameter is 100cm, the temperature of the culture water is 20-25 ℃, and the dissolved oxygen is 6-7mg/L, ph 7.8.8-8.4.
4. The method according to claim 1, wherein the composite polysaccharide nanoparticle in the step (1) is added at a concentration of 0.02-0.3g/L, and the composite probiotics are lactic acid bacteria and bacillus subtilis, and the composite probiotics are added at a concentration of 0.1-0.3g/L.
5. The method according to claim 2, wherein the mass ratio of chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide is 1-12:3-23:1.5-6:1-8:8-30 parts; the concentration of polysaccharide solutions prepared from chitosan, astragalus polysaccharide, xylooligosaccharide, rosa roxburghii polysaccharide and lycium barbarum polysaccharide are respectively as follows: 10-120mg/L, 30-230mg/L, 15-60mg/L, 10-80mg/L and 80-300mg/L.
6. The method of claim 2, wherein the ultrasonic dispersion has an ultrasonic time of 10 to 30 minutes and a power of 40 to 60W.
7. The method of claim 1, wherein the transport temperature is 20-25 ℃.
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