CN107455615B - Artemia nutrition strengthening method suitable for culturing octopus ocellatus planktonic larvae - Google Patents
Artemia nutrition strengthening method suitable for culturing octopus ocellatus planktonic larvae Download PDFInfo
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- CN107455615B CN107455615B CN201710597565.6A CN201710597565A CN107455615B CN 107455615 B CN107455615 B CN 107455615B CN 201710597565 A CN201710597565 A CN 201710597565A CN 107455615 B CN107455615 B CN 107455615B
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Abstract
The invention discloses a artemia nutrition strengthening method suitable for culturing octopus ocellatus planktonic larvae, which comprises the following steps: preparing tea polyphenol liposome, extracting fish visceral oil, modifying fish oil and enhancing nutrition. The beneficial effects are that: by extracting fish visceral oil and carrying out enzymatic modification, saturated fatty acid in the fish oil can be converted into unsaturated fatty acid, the content of unsaturated fatty acid such as DHA, EPA and the like in artemia bodies can be greatly improved, the nutrition of the reinforced artemia is more comprehensive and balanced, the nutrition requirement for culturing octopus ocellatus planktonic larvae can be met, the lipid and fatty acid composition of larvae can be changed, the growth speed, the survival rate, the immunity and the stress resistance of the larvae are improved, and therefore the method is an efficient and safe artemia nutrition reinforcing method suitable for culturing octopus ocellatus planktonic larvae.
Description
Technical Field
The invention relates to the field of aquatic larva culture, in particular to a artemia nutrition strengthening method suitable for octopus ocellatus planktonic larva culture.
Technical Field
The real kurtosis is one of the kurtosis animals with higher economic value, is one of the cephalopods which are most researched and have the highest popularity in the world, and has wide markets in the Mediterranean, Latin America and Asia. The true kurtosis has the following characteristics suitable for cultivation: 1) the food conversion rate is high, and the food can be converted to 40-60% of the digested food; 2) the growth rate is high, and the growth rate is about 3% per day; 3) the protein content is high and accounts for 70-90% of dry weight; 4) the amount of the eggs is high, and each mature individual has 10-50 ten thousand eggs. Due to the wide market space and excellent biological characters, a plurality of scholars conduct extensive researches on the ecology, physiology, behavior, reproduction, growth and cultivation of the scholars for years, and a good foundation is provided for developing the true kurtosis cultivation. The length of the body of the true kurtosis larva is 1-1.5mm when the true kurtosis larva is just hatched, the true kurtosis larva moves actively, the metabolism rate is high, and a large amount of living body bait with rich nutrition is needed. In the planktonic stage, their morphologies change significantly, and the palpation grows faster than the popliteal part, and can change from the cuttlefish shape during hatching to the adult shape. Bait is a crucial factor in larval rearing. Researchers are based on the fat composition of larvae and foods thereof, and suggest that foods rich in polyunsaturated fatty acids, especially 2 foods of DHA and EPA, are really needed, which is an important factor for measuring the high and low quality of the bait.
The artemia serving as an important food organism and a good experimental animal material is widely paid attention by people all the time, nauplii of the artemia serve as the best choice for the first meal of a plurality of tropical aquatic organisms, in the cultivation practice, the artemia serves as a very ideal octopus ocellatus planktonic larva bait, and researches show that the artemia has good palatability and obvious feeding effect, but biochemical components of the artemia lack PUFA and cannot completely meet the nutritional requirements of the octopus ocellatus, so that the octopus ocellatus planktonic larva has high death rate and low growth speed, and the strengthening of the octopus ocellatus planktonic larva bait becomes particularly important.
At present, few technical methods for culturing and enriching nutrition of octopus ocellatus planktonic larvae are provided, and the prior art, such as a Chinese patent invention with an authorized public number of CN 103141439B, discloses a method for enriching DHA, EPA and ARA in artemia, wherein the method is used for enriching nutrition of the artemia by feeding the artemia after superfine grinding of euphausia superba powder, and has the advantages of simple operation and low cost.
Disclosure of Invention
The invention aims to provide an efficient and safe artemia nutrition strengthening method suitable for culturing octopus ocellatus planktonic larvae.
Aiming at the problems mentioned in the background technology, the invention adopts the technical scheme that: a method for enriching the nutrients of artemia suitable for culturing the planktonic larvae of octopus ocellatus includes the following steps:
preparing tea polyphenol liposome: weighing 850 parts of mixed solution of 7.5-7.8 parts of sodium dihydrogen phosphate and disodium hydrogen phosphate of 0.04-0.05mol/L, pH, slowly adding 8.0-8.1 parts of tea polyphenol, heating to 58-60 ℃ under strong stirring, and keeping constant temperature; weighing 300-320 parts of yolk lecithin, 58-60 parts of cholesterol and 16-17 parts of VE, adding the mixture into 600 parts of anhydrous ethanol, heating the mixture to 58-60 ℃, stirring and dissolving the mixture, adding the mixed solution into a tea polyphenol solution after the mixture is completely dissolved, maintaining the temperature and stirring the mixture for 30-45 minutes, performing rotary evaporation for 20-25 minutes at the temperature of 55-60 ℃ and under the vacuum of 0.1-0.15MPa to remove residual ethanol, performing ultrasonic treatment with the ultrasonic power of 200-230W and the frequency of 23-25KHz, performing ultrasonic treatment for 120-180 seconds at the frequency of 4 seconds at intervals of 4 seconds of ultrasonic 4 seconds, and storing the tea polyphenol liposome at the temperature of 3-4 ℃ after the ultrasonic treatment; the tea polyphenol liposome prepared by the method has high encapsulation rate, is easy to be absorbed by artemia, has simple preparation method, avoids the physicochemical treatment of potential harmful substances such as diethyl ether and the like in the preparation process, and is very suitable for industrialized large-scale production;
extracting fish visceral oil: taking fresh fish viscera, removing digestion residues in the viscera, cleaning, draining, homogenizing in a homogenizer, weighing 210 parts of the fish viscera, homogenizing in a beaker, adding 60-65 parts of distilled water, heating to 30-34 ℃ in a water bath under stirring, adjusting the pH to 8-9, continuously stirring, heating to 45-55 ℃, keeping the temperature for 25-35 minutes, adding potassium chloride accounting for 5-7% of the weight of the fish viscera, stirring, dissolving, salting out for 20-30 minutes, centrifuging while hot, and separating to obtain crude fish oil; extracting fish visceral oil by a potassium method to remove impurities, and preparing for further modification; the method has high fish oil yield and simple operation, and the waste water and the waste residue can be prepared into green high-efficiency fertilizer to be fully and comprehensively utilized and can also be used as raw materials for extracting nucleic acid to be further high-valued, thereby achieving the purpose of comprehensively utilizing wastes;
modifying the fish oil: weighing 110 parts of crude fish oil and 0.07-0.08 part of vitamin C, putting the crude fish oil and the vitamin C into a closed container, uniformly mixing, putting the reaction container into an ultrasonic instrument, heating to 70-73 ℃, keeping the temperature, continuously introducing oxygen for 5-8 minutes, then adding 0.3-0.4 part of desaturase and 0.05-0.06 part of reduced coenzyme II, and reacting for 140 seconds under the action of high-ultrasonic waves with the power of 400-500W and the frequency of 80-90KHz to obtain the crude fish oil and the vitamin C for later use; performing siphon enzymatic on the fish oil, washing with hot water for 5-3 times, centrifuging while hot, collecting the upper layer liquid, and drying in a vacuum drying oven to obtain fish visceral oil; the enzymatic reaction can deprive hydrogen of a carbon-carbon double bond at the middle position of saturated fatty acid by oxygen to generate water, so that long-chain saturated fatty acid loses hydrogen and is converted into corresponding unsaturated fatty acid, the fish visceral oil is rich in polyunsaturated fatty acid such as DHA (docosahexaenoic acid), EPA (eicosapentaenoic acid) and the like and is very suitable for serving as an artemia reinforcing component, and when the mass ratio of the desaturase to the reduced coenzyme is 6-7:1, the reaction activity is highest;
and (3) nutrition fortification: weighing 10-15 parts of schizochytrium limacinum, 20-25 parts of tea polyphenol liposome, 20-25 parts of fish visceral oil, 28-33 parts of fish protein hydrolysate, 10-15 parts of chlorella, 20-23 parts of urea, 18-20 parts of ammonium nitrate, 15-20 parts of calcium phosphate, 60-80 parts of fertilizer and 2-4 parts of phagostimulant, fully and uniformly mixing, crushing into particles with the size of 10-45 microns to obtain nutrition-enhanced bait, feeding 4-6g of bait with 10g of polypide to enhance artemia salina, diluting the bait in a small amount of pool water, uniformly sprinkling the bait in the whole pool, feeding once every 2-3 hours, wherein the feeding amount is equal every time, and feeding the octopus ocellatus plankton larvae after enhancing feeding for 20-24 hours; the reinforced bait is rich in HUFA, protein and other nutrient substances such as DHA, EPA and the like, can promote the digestion and absorption of artemia under the combined action of the feeding promoting components and the fertilizer, so that the artemia can accelerate the growth, the saturated fatty acid content in the bait is extremely low, the survival rate of the artemia is greatly improved compared with that of the artemia fed by common fish oil bait, the HUFA content of the artemia such as EPA, DHA and the like is obviously improved after the reinforced artemia is reinforced, and the lipid and fatty acid composition of the artemia can be changed after the enriched artemia is fed to octopus ocellatus planktonic larvae, so that the growth speed, the survival rate, the immunity and the stress resistance of the larvae are improved, therefore, the method is an effective artemia nutrition reinforcing method.
Preferably, the fish is weever, bighead carp or silver carp, the viscera of the fish are mostly discarded, and the fatty acid content of the fish is high.
Preferably, the mass ratio of desaturase to reduced coenzyme used is 6-7:1, and the enzymatic reaction proceeds most thoroughly while maintaining this ratio.
Preferably, the fertilizer is cow dung or chicken dung or pig dung or bean cake or cottonseed meal or rapeseed meal, and effectively provides organic matters and medium and trace elements such as nitrogen, phosphorus, potassium, calcium, iron, magnesium and the like for the artemia.
Preferably, the used phagostimulant is citrus peel powder, taurine, yeast powder, silkworm chrysalis powder or brewer's grain, and the natural phagostimulant is helpful for promoting the digestion and absorption of the artemia.
Compared with the prior art, the invention has the advantages that: 1) the method uses waste fish viscera produced in fishery as raw materials to extract fish visceral oil, accurately controls enzymatic reaction conditions, and converts saturated fatty acid in the visceral oil into unsaturated fatty acid, thereby not only reducing the harm of the saturated fatty acid to artemia, but also greatly improving the content of the unsaturated fatty acid in bait, and being suitable for being used as the bait for strengthening the artemia; 2) the method can greatly improve the content of HUFAs such as DHA, EPA and the like in artemia bodies, enables the nutrition of the reinforced artemia to be more comprehensive and balanced, can meet the nutritional requirement of culture of octopus ocellatus planktonic larvae, can change the lipid and fatty acid composition of the larvae, and improves the growth speed, survival rate, immunity and stress resistance of the larvae, so the nutrition-reinforced formula is an efficient and safe nutrition-reinforced formula suitable for culture of octopus ocellatus planktonic larvae.
Drawings
FIG. 1 is a graph showing the survival and weight gain of artemia at various boosting times for examples 1, 2 and 3 of the present invention and a control group;
FIG. 2 is a graph showing the survival rate of planktonic larvae of Octopus vulgaris in different time periods according to the present invention;
FIG. 3 is a graph showing the carcass length growth rate of octopus ocellatus planktonic larvae at different time periods in the present invention;
FIG. 4 is a graph showing the content of unsaturated fatty acids in the fish oil of examples 1 to 3 of the present invention.
Detailed Description
The scheme of the invention is further illustrated by the following examples:
example 1:
a method for enriching the nutrients of artemia suitable for culturing the planktonic larvae of octopus ocellatus includes the following steps:
1) preparing a phosphate solution of tea polyphenol, weighing 300 parts of egg yolk lecithin, 58 parts of cholesterol and 16 parts of VE, adding the mixture into 500 parts of absolute ethyl alcohol, heating the mixture to 58 ℃, stirring and dissolving the mixture, adding the mixed solution into the tea polyphenol solution after complete dissolution, maintaining the temperature, stirring the mixture for 30 minutes, performing rotary evaporation at the temperature of 55 ℃ to remove residual ethanol, performing ultrasonic treatment, and storing the tea polyphenol liposome at the temperature of 3 ℃; the tea polyphenol liposome prepared by the method has high encapsulation rate and is easy to be absorbed by artemia; 2) weighing 200 parts of fresh weever viscera, homogenizing the fresh weever viscera into a beaker, adding 60 parts of distilled water, heating the water bath to 30 ℃ under stirring, adjusting the pH to 8, continuing stirring and heating to 45 ℃, keeping the temperature for 25 minutes, adding 10 parts of potassium chloride, stirring and dissolving, salting out for 20 minutes, centrifuging the mixture while the mixture is hot, and separating to obtain crude fish oil; 3) weighing 100 parts of crude fish oil and 0.07 part of vitamin C, putting the crude fish oil and the vitamin C into a closed container, uniformly mixing, putting the reaction container into an ultrasonic instrument, heating to 70 ℃, keeping the temperature, continuously introducing oxygen for 5 minutes, then adding 0.3 part of desaturase and 0.05 part of reduced coenzyme II, and reacting for 110 seconds under the action of high ultrasonic waves for later use; performing siphon enzymatic on the fish oil, washing with hot water for 5 times, centrifuging while hot, taking the upper layer liquid, and drying in a vacuum drying oven to obtain fish visceral oil; the enzymatic reaction can deprive hydrogen of a carbon-carbon double bond at the middle position of saturated fatty acid by oxygen to generate water, so that long-chain saturated fatty acid loses hydrogen and is converted into corresponding cis-form unsaturated fatty acid, and the fish visceral oil is rich in polyunsaturated fatty acid such as DHA (docosahexaenoic acid), EPA (eicosapentaenoic acid) and the like; 4) weighing 10 parts of schizochytrium limacinum, 20 parts of tea polyphenol liposome, 20 parts of fish visceral oil, 28 parts of fish protein hydrolysate, 10 parts of chlorella, 20 parts of urea, 18 parts of ammonium nitrate, 15 parts of calcium phosphate, 60 parts of peanut meal and 2 parts of silkworm chrysalis meal, fully and uniformly mixing, crushing into 10-micron particles to obtain nutrition-enhanced bait, feeding 4g of the bait with 10g of artemia at a feed rate, feeding once every 3 hours with the same feed rate, and feeding the octopus ocellatus planktonic larvae after 21 hours of enhanced feeding; the reinforced bait is rich in HUFA such as DHA, EPA and the like, protein and other nutrient substances, and can promote the digestion and absorption of the artemia under the combined action of the feeding promoting ingredients and the fertilizer, so that the artemia can accelerate growth.
Example 2:
a method for enriching the nutrients of artemia suitable for culturing the planktonic larvae of octopus ocellatus includes the following steps:
1) 850 parts of phosphate solution with the tea polyphenol content of 1% is prepared, and the phosphate solution is heated to 60 ℃ under strong stirring and kept at a constant temperature; weighing 320 parts of egg yolk lecithin, 60 parts of cholesterol and 17 parts of VE, adding the egg yolk lecithin, the cholesterol and the VE into 600 parts of absolute ethyl alcohol, heating to 60 ℃, stirring for dissolving, adding the mixed solution into a tea polyphenol solution after complete dissolution, maintaining the temperature, stirring strongly for 45 minutes, carrying out rotary evaporation for 25 minutes at the temperature of 60 ℃ and under the vacuum of 0.15MPa to remove residual ethanol, and storing the tea polyphenol liposome at the temperature of 3-4 ℃ after the ultrasonic treatment is finished; the preparation method of the tea polyphenol liposome is simple, physical and chemical treatment of potential harmful substances such as ether and the like is avoided in the preparation process, and the tea polyphenol liposome is very suitable for industrial large-scale production;
2) weighing 210 parts of fresh chub viscera, homogenizing in a beaker, adding 65 parts of distilled water, heating the mixture in a water bath to 34 ℃ under stirring, adjusting the pH to 9, continuing stirring and heating the mixture to 55 ℃, keeping the temperature for 35 minutes, adding 14.7 parts of potassium chloride, stirring and dissolving, salting out for 30 minutes, centrifuging while hot, and separating to obtain crude fish oil; extracting fish visceral oil by a potassium method to remove impurities, and preparing for further modification;
3) weighing 110 parts of crude fish oil and 0.08 part of vitamin C, putting the crude fish oil and the vitamin C into a closed container, uniformly mixing, putting the reaction container into an ultrasonic instrument, heating to 73 ℃, keeping the temperature, continuously introducing oxygen for 8 minutes, then adding 0.4 part of desaturase and 0.06 part of reduced coenzyme II, reacting for 140 seconds under the action of high ultrasonic waves, siphoning enzymatic fish oil, washing the enzymatic fish oil for 3 times by hot water, centrifuging the fish oil while hot, taking upper-layer liquid, and drying the liquid by using a vacuum drying oven to obtain fish visceral oil; the method has the advantages that the content of unsaturated fatty acid in the fish oil is high, the operation is simple, the waste water and waste residue can be prepared into green high-efficiency fertilizer to be fully and comprehensively utilized, and the waste water and waste residue can be used as raw materials for extracting nucleic acid to be further high-valued, so that the aim of comprehensively utilizing waste is fulfilled;
4) weighing 15 parts of schizochytrium limacinum, 25 parts of tea polyphenol liposome, 25 parts of fish visceral oil, 33 parts of fish protein hydrolysate, 15 parts of chlorella, 23 parts of urea, 20 parts of ammonium nitrate, 20 parts of calcium phosphate, 40 parts of cow dung, 40 parts of rapeseed meal and 4 parts of brewer's grain, fully and uniformly mixing, crushing into particles of 45 micrometers to obtain nutrition-enriched bait, feeding 6g of the bait with 10g of polypide to enhance artemia salina, diluting the bait in a small amount of pond water during feeding, uniformly sprinkling the bait in the whole pond, feeding once every 3 hours, wherein the feeding amount is equal every time, and feeding the octopus ocellatus planktonic larvae after 24 hours of enhanced feeding; the contents of HUFAs such as EPA, DHA and the like in the reinforced artemia are obviously improved, the immunity of the artemia is enhanced, the nutritive value is richer, and after the reinforced artemia is fed to octopus ocellatus planktonic larvae, the composition of lipid and fatty acid of the planktonic larvae can be changed, the growth speed, the survival rate, the immunity and the stress resistance of the larvae are improved, so the method is an effective artemia nutrition reinforcing method.
Example 3:
a method for enriching the nutrients of artemia suitable for culturing the planktonic larvae of octopus ocellatus includes the following steps:
preparing tea polyphenol liposome: weighing 840 parts of mixed solution of 7.8 parts of sodium dihydrogen phosphate and disodium hydrogen phosphate at 0.05mol/L, pH, slowly adding 8.0 parts of tea polyphenol, heating to 58 ℃ under strong stirring, and keeping the temperature constant; weighing 300 parts of yolk lecithin, 58 parts of cholesterol and 17 parts of VE, adding the yolk lecithin, 58 parts of cholesterol and 17 parts of VE into 580 parts of absolute ethyl alcohol, heating to 60 ℃, stirring for dissolving, adding the mixed solution into a tea polyphenol solution after complete dissolution, maintaining the temperature, stirring strongly for 40 minutes, carrying out rotary evaporation for 25 minutes at the temperature of 60 ℃ and under the vacuum of 0.12MPa to remove residual ethanol, then carrying out ultrasonic treatment, wherein the ultrasonic power is 220W, the frequency is 24 KHz, carrying out ultrasonic treatment for 160 seconds at the frequency of 4 seconds at intervals of ultrasonic treatment, and preserving the tea polyphenol liposome at the temperature of 3 ℃ after the ultrasonic treatment is finished; the tea polyphenol liposome prepared by the method has high encapsulation rate, is easy to be absorbed by artemia, has simple preparation method, avoids the physicochemical treatment of potential harmful substances such as diethyl ether and the like in the preparation process, and is very suitable for industrialized large-scale production;
extracting fish visceral oil: taking fresh weever viscera, removing digestion residues in the viscera, cleaning, draining, homogenizing in a homogenizer, weighing 210 parts of the viscera, homogenizing in a beaker, adding 60 parts of distilled water, heating the water bath to 33 ℃ under stirring, adjusting the pH to 9, continuously stirring, heating to 50 ℃, keeping the temperature for 30 minutes, adding 12 parts of potassium chloride, stirring for dissolving, salting out for 25 minutes, centrifuging while hot, and separating to obtain crude fish oil; extracting fish visceral oil by a potassium method to remove impurities, and preparing for further modification; the method has high fish oil yield and simple operation, and the waste water and the waste residue can be prepared into green high-efficiency fertilizer to be fully and comprehensively utilized and can also be used as raw materials for extracting nucleic acid to be further high-valued, thereby achieving the purpose of comprehensively utilizing wastes;
modifying the fish oil: weighing 0.16g of vitamin E, 0.50g of ethylenediamine tetraacetic acid, 0.14g of thio-lauric anhydride, 0.02g of L- (+) -2, 3-dihydroxypropionic acid, 2.3g of acrylic resin modified casein, 2.5g of polyurethane modified nitrocellulose and 11g of propylene glycol ether in sequence, placing the mixture in a high-speed mixer at 8000r/min, mixing for 3 minutes to obtain a coating preparation, uniformly coating the coating preparation on the inner bottom surface and the inner wall of a closed reaction container, and drying for 25 minutes at the temperature of 25 ℃ for later use; weighing 100 parts of crude fish oil and 0.08 part of vitamin C, putting into a closed container, uniformly mixing, and placing a reaction container into the closed containerHeating to 72 ℃ in an ultrasonic instrument, keeping the temperature, continuously introducing oxygen for 6 minutes, then adding 0.4 part of desaturase and 0.06 part of reduced coenzyme II, and reacting for 130 seconds under the action of high ultrasonic waves with the power of 450W and the frequency of 87KHz for later use; performing siphon enzymatic on the fish oil, washing with hot water for 4 times, centrifuging while hot, taking the upper layer liquid, and drying in a vacuum drying oven to obtain fish visceral oil; the film-forming components in the coating preparation have synergistic effect, so that the coating property is more stable, the coating texture is more uniform, the permeability of the film is more uniform, the release of effective substances in the film is facilitated, and the effective substances can react with Mg2+、Ca2+、Mn2+、Zn2+、Fe2+、Al3+Combining the secondary and trivalent metal ions, thereby reducing the activity inhibition of the metal ions on the reduced coenzyme II and the desaturase; particularly, when the mass ratio of the thio-lauric anhydride to the L- (+) -2, 3-dihydroxypropionic acid is 7:1, the product of the enzymatic reaction is mainly cis-unsaturated fatty acid, the generation rate of trans-unsaturated fatty acid is reduced, and the toxic and side effects of the unsaturated fatty acid on organisms are reduced; the effective substances released by the coating also have the effect of removing redundant free radicals in the fish oil, so that the oxidative degradation of the free radicals to cis-unsaturated fatty acid is minimized, and the stability of enzymatic products is enhanced; the enzymatic reaction can deprive hydrogen of a carbon-carbon double bond at the middle position of saturated fatty acid by oxygen to generate water, so that long-chain saturated fatty acid loses hydrogen and is converted into corresponding cis-form unsaturated fatty acid, and the fish visceral oil is rich in polyunsaturated fatty acid such as DHA (docosahexaenoic acid), EPA (eicosapentaenoic acid) and the like and is very suitable for serving as an artemia strengthening component;
and (3) nutrition fortification: weighing 12 parts of schizochytrium limacinum, 24 parts of tea polyphenol liposome, 24 parts of fish visceral oil, 30 parts of fish protein hydrolysate, 10 parts of chlorella, 20 parts of urea, 20 parts of ammonium nitrate, 16 parts of calcium phosphate, 30 parts of chicken manure, 40 parts of cottonseed meal and 3 parts of yeast powder, fully mixing uniformly, crushing into particles with the size of 30 microns to obtain nutrition-enhanced bait, feeding 5g of the bait with 10g of polypide to enhance artemia salina, diluting the bait in a small amount of pond water during feeding, uniformly sprinkling the bait in the whole pond, feeding once every 2 hours, wherein the feeding amount is equal every time, and feeding the octopus ocellatus planktonic larvae after 24 hours of enhanced feeding; the reinforced bait is rich in HUFA, protein and other nutrient substances such as DHA, EPA and the like, can promote the digestion and absorption of artemia under the combined action of the feeding promoting components and the fertilizer, so that the artemia can accelerate the growth, the saturated fatty acid content in the bait is extremely low, the survival rate of the artemia is greatly improved compared with that of the artemia fed by common fish oil bait, the HUFA content of the artemia such as EPA, DHA and the like is obviously improved after the reinforced artemia is reinforced, and the lipid and fatty acid composition of the artemia can be changed after the enriched artemia is fed to octopus ocellatus planktonic larvae, so that the growth speed, the survival rate, the immunity and the stress resistance of the larvae are improved, therefore, the method is an effective artemia nutrition reinforcing method.
Control group:
crushing small fish feed purchased from Tianjin Chenghui feed Co., Ltd into 45-micron particles, feeding artemia with the feed amount of 1/2, diluting the bait in a small amount of pool water during feeding, uniformly sprinkling the bait in the whole pool, feeding once every 2 hours with the same feed amount, and feeding octopus ocellatus planktonic larvae after feeding for 22 hours.
The average values of the survival rate and the weight gain rate of the reinforced artemia are measured respectively when the artemia are fed for different time periods, the experimental data are shown in figure 1, and as can be seen from figure 1, the reinforced artemia of examples 1-3 are superior to a control group in the aspects of the survival rate and the weight gain rate, so that the nutrition of the artemia nutrition reinforced formula disclosed by the invention is more comprehensive and balanced, the immunity, the antibacterial and disease-resistant capability of the artemia can be obviously improved after the artemia is fed, and the survival rate of the artemia is higher.
The artemia strengthened in the examples 1, 2 and 3 and the control group are fed to octopus vulgaris planktonic larvae, under the same feeding condition, the survival rate and the carcass length increase rate of the octopus vulgaris planktonic larvae in different time periods are taken for data analysis, as shown in fig. 2 and 3, as can be seen from fig. 2 and 3, the examples 1-3 are obviously superior to the control group in the aspects of the survival rate and the growth rate of the octopus vulgaris planktonic larvae, and along with the prolonging of the feeding time, the larva death rate in the examples 1-3 can be gradually reduced, the growth rate is stably increased, and the artemia strengthened by management and cultivation are proved to be highly effective in the aspects of improving the growth rate, the survival rate, the immunity, the stress resistance and the like of the larvae.
Respectively measuring the content of fatty acid in the fish oil obtained in the embodiment 1-3, wherein the content of unsaturated fatty acid is determined by measuring the iodine value, and the specific measuring method of the iodine value can be found in GB/T5532-2008; trans-unsaturated fatty acid has an absorption peak at a specific position of an infrared spectrogram, cis-unsaturated fatty acid does not have the characteristic, a linear regression equation is established by utilizing the peak area of a characteristic peak and the corresponding concentration of the characteristic peak, and the trans-unsaturated fatty acid can be quantitatively analyzed, so that the content of the cis-unsaturated fatty acid is obtained, as shown in fig. 4. As can be seen from FIG. 4, after purification and desaturation enzymatic reaction, the content of unsaturated fatty acids in the fish oil of examples 1-3 is much higher than that of the control group, and after the coating is added to the reaction vessel for enzymatic reaction in example 3, not only the content of unsaturated fatty acids in the fish oil is increased, but also the content of cis-unsaturated fatty acids is significantly increased, which indicates that the coating not only promotes the generation of cis-unsaturated fatty acids, but also is beneficial to desaturation of saturated fatty acids; after the artemia is fed intensively, the content of polyunsaturated fatty acid in the artemia can be greatly improved, so that the artemia is more comprehensive and balanced in nutrition, the nutrition requirement for culturing octopus ocellatus planktonic larvae can be met, the lipid and fatty acid composition of larvae can be changed, the growth speed, the survival rate, the immunity and the stress resistance of larvae are improved, and therefore the method has important significance for strengthening the artemia and further culturing the octopus ocellatus planktonic larvae.
Conventional operations in the operation steps of the present invention are well known to those skilled in the art and will not be described herein.
The embodiments described above are intended to illustrate the technical solutions of the present invention in detail, and it should be understood that the above-mentioned embodiments are only specific embodiments of the present invention, and are not intended to limit the present invention, and any modification, supplement or similar substitution made within the scope of the principles of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A method for enriching the nutrients of artemia suitable for culturing the planktonic larvae of octopus ocellatus includes the following steps: the preparation method of the tea polyphenol liposome comprises the steps of preparation of the tea polyphenol liposome, extraction of fish visceral oil, modification of fish oil and nutrition enhancement, and is characterized in that in the step of modification of the fish oil, desaturase and reduced coenzyme II are used for carrying out enzymatic modification on the fish oil.
2. The method for enriching the nutrition of artemia salina suitable for the culture of octopus ocellatus planktonic larvae as claimed in claim 1, wherein: the preparation steps of the tea polyphenol liposome are as follows: weighing 850 parts of mixed solution of 7.5-7.8 parts of sodium dihydrogen phosphate and disodium hydrogen phosphate of 0.04-0.05mol/L, pH, slowly adding 8.0-8.1 parts of tea polyphenol, heating to 58-60 ℃ under strong stirring, and keeping constant temperature; weighing 300-320 parts of yolk lecithin, 58-60 parts of cholesterol and 16-17 parts of VE, adding the mixture into 600 parts of anhydrous ethanol, heating to 58-60 ℃, stirring and dissolving, adding the mixed solution into a tea polyphenol solution after complete dissolution, maintaining the temperature and stirring strongly for 30-45 minutes, performing rotary evaporation for 20-25 minutes at the temperature of 55-60 ℃ and under the vacuum of 0.1-0.15MPa to remove residual ethanol, performing ultrasonic treatment at the ultrasonic power of 200-230W and the frequency of 23-25KHz, performing ultrasonic treatment for 2-3 minutes at the ultrasonic frequency of 4 seconds, and storing the tea polyphenol liposome at the temperature of 3-4 ℃ after the ultrasonic treatment is finished.
3. The method for enriching the nutrition of artemia salina suitable for the culture of octopus ocellatus planktonic larvae as claimed in claim 1, wherein: the method for extracting the fish visceral oil comprises the following steps: taking fresh fish viscera, removing digestion residues in the viscera, cleaning, draining, homogenizing in a homogenizer, weighing 210 parts of the fish viscera, homogenizing in a beaker, adding 60-65 parts of distilled water, heating to 30-34 ℃ in a water bath under stirring, adjusting the pH to 8-9, continuously stirring, heating to 45-55 ℃, keeping the temperature for 25-35 minutes, adding potassium chloride accounting for 5-7% of the weight of the fish viscera, stirring, dissolving, salting out for 20-30 minutes, centrifuging while hot, and separating to obtain crude fish oil.
4. The method for enriching the nutrition of artemia salina suitable for the culture of octopus ocellatus planktonic larvae as claimed in claim 1, wherein: the fish used in the step of extracting the fish visceral oil comprises the following steps: weever, bighead carp or silver carp.
5. The method for enriching the nutrition of artemia salina suitable for the culture of octopus ocellatus planktonic larvae as claimed in claim 1, wherein: the fish oil modification steps are as follows: weighing 110 parts of crude fish oil and 0.07-0.08 part of vitamin C, putting the crude fish oil and the vitamin C into a closed container, uniformly mixing, putting the reaction container into an ultrasonic instrument, heating to 70-73 ℃, keeping the temperature, continuously introducing oxygen for 5-8 minutes, then adding 0.3-0.4 part of desaturase and 0.05-0.06 part of reduced coenzyme II, and reacting for 140 seconds under the action of high-ultrasonic waves with the power of 400-500W and the frequency of 80-90KHz to obtain the crude fish oil and the vitamin C for later use; and (3) siphoning enzymatic fish oil, washing with hot water for 5-3 times, centrifuging while hot, taking the upper layer liquid, and drying in a vacuum drying oven to obtain the fish visceral oil.
6. The method for enriching the nutrition of artemia salina suitable for the culture of octopus ocellatus planktonic larvae as claimed in claim 1, wherein: in the step of modifying the fish oil, the mass ratio of the desaturase to the reduced coenzyme II is 6-7: 1.
7. The method for enriching the nutrition of artemia salina suitable for the culture of octopus ocellatus planktonic larvae as claimed in claim 1, wherein: the nutrition enhancing step comprises the following steps: weighing 10-15 parts of schizochytrium limacinum, 20-25 parts of tea polyphenol liposome, 20-25 parts of fish visceral oil, 10-15 parts of chlorella, 30-40 parts of fertilizer and 2-4 parts of phagostimulant, fully and uniformly mixing, crushing into particles with the size of 10-45 microns to obtain nutrition-enhanced bait, feeding 4-6g of the bait with 10g of polypide to enhance artemia, diluting the bait in a small amount of pool water during feeding, uniformly sprinkling the bait in the whole pool, feeding once every 2-3 hours, wherein the feeding amount is equal every time, and feeding the octopus ocellatus planktonic larvae after 20-24 hours of enhanced feeding.
8. The method for enriching the nutrition of artemia salina suitable for the culture of octopus ocellatus planktonic larvae as claimed in claim 7, wherein: the fertilizer is cow dung or chicken dung or pig dung or bean cake or cottonseed meal or rapeseed meal, and the phagostimulant is citrus peel powder or taurine or yeast powder or silkworm chrysalis powder or brewer's grains.
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