CN113016678A - Living body microalgae composition and application thereof in feeding nutrition-enhanced artemia and litopenaeus vannamei - Google Patents
Living body microalgae composition and application thereof in feeding nutrition-enhanced artemia and litopenaeus vannamei Download PDFInfo
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- CN113016678A CN113016678A CN202110246398.7A CN202110246398A CN113016678A CN 113016678 A CN113016678 A CN 113016678A CN 202110246398 A CN202110246398 A CN 202110246398A CN 113016678 A CN113016678 A CN 113016678A
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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
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
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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
- A01K61/50—Culture of aquatic animals of shellfish
- A01K61/59—Culture of aquatic animals of shellfish of crustaceans, e.g. lobsters or shrimps
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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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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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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Abstract
The invention discloses a living microalgae composition for feeding artemia, daphnia larvae of litopenaeus vannamei and shrimps, which comprises the following components: the inventor finds that the value of brine shrimp and young shrimp can be greatly improved when the fine horn hair microalgae and the schizochytrium limacinum are combined and fed to the young shrimp for feeding brine shrimp and litopenaeus vannamei, the metamorphosis rate and the survival rate of the daphnia larval can be obviously improved when the daphnia larval of the litopenaeus vannamei is fed, malondialdehyde, total antioxidant capacity, alkaline phosphatase and acid phosphatase activity are obviously improved, and discloses a specific application method of the living body microalgae composition in feeding the brine shrimp and the litopenaeus vannamei.
Description
Technical Field
The invention relates to the technical field of prawn feeding, in particular to a living microalgae composition and application thereof in nutrition-enhanced artemia, in the feeding of daphnia-shaped larvae of litopenaeus vannamei, and in the feeding of litopenaeus vannamei.
Background
The litopenaeus vannamei (Litopenaeus vannamei), commonly known as Penaeus vannamei, belongs to Arthropoda and Penaeus (Penaeus), has a shape similar to that of Penaeus chinensis (Fenneropenaeus chinensis), has thin shell, normal body color of light grey, and no speckle throughout the body. The litopenaeus vannamei is native to the middle of the pacific coast to the west coast, in gulf of mexico, has the advantages of high growth speed, strong stress resistance, high feed utilization rate, high meat yield, remarkable culture economic benefit and the like, and is called three high-quality varieties with highest culture yield in the world together with penaeus monodon (Penaeusmonodon) and Chinese prawns.
Since the introduction of the litopenaeus vannamei into China in 1988, the litopenaeus vannamei is gradually bred and popularized in large scale in coastal areas in south China. In 2013 and 2018, the total yield of the litopenaeus vannamei culture in China is increased year by year, accounts for over 70 percent of the shrimp culture yield, and is the breed with the widest range and the highest culture yield in China.
In the process of breeding the litopenaeus vannamei, along with the rise of breeding density, the problems of germplasm resource degradation, water quality deterioration, difficult shrimp seedling breeding, uneven feed quality and the like are caused, the healthy and continuous development of the litopenaeus vannamei breeding industry is seriously restricted, the growth and disease resistance of the litopenaeus vannamei larvae of different strains are different under different environmental conditions, the litopenaeus vannamei larvae are more active in the stage from the daphnia stage to the mysid stage, the growth is fast, and the litopenaeus vannamei larvae are easily influenced by the external environment and invaded by pathogenic bacteria, so that the survival rate of the. The problem that the seedling production is affected is common because proper initial baits are not fed in proper time, and the growth of the prawns is affected because most baits on the market have single nutrient components. Therefore, it is necessary to research and screen high-quality bait suitable for prawn growth.
Disclosure of Invention
In order to solve at least one technical defect, the invention provides the following technical scheme:
the first aspect of the present document discloses a live microalgae composition for feeding artemia, daphnia larvae of litopenaeus vannamei, and shrimp, comprising the following components: fine horn hair microalgae, schizochytrium limacinum.
The inventor finds that the values of the artemia and the young shrimps can be greatly improved when the gracillin microalgae and the schizochytrium limacinum are fed to the young shrimps for feeding the artemia and the litopenaeus vannamei in a combined mode, and the metamorphosis rate and the survival rate of the daphnia larvas, Malondialdehyde (MDA), total antioxidant capacity (T-AOC), alkaline phosphatase (AKP) and acid phosphatase (ACP) activities can be remarkably improved when the daphnia larvas are fed.
For the component proportion of the tenuifolius glauca microalgae and the schizochytrium limacinum, 1:1, 1:2,2:1, 3:1,1:3, and the like.
The second aspect of the application document discloses an application of the living microalgae composition in breeding of daphnoid larvae of litopenaeus vannamei, wherein the daphnoid larvae of litopenaeus vannamei are cultivated in a cultivation box containing seawater, the dissolved oxygen amount of the seawater is kept to be more than 6mg/L during the cultivation process, the water temperature is 25-30 ℃, the living composition of the tenuifolius cerasus microalgae and the schizochytrium limacinum is fed when the larvae begin to become daphnoid in the first period, and the feeding density is 2-4 x 104cells/ml。
The difference of the traditional scheme is that the combination of the micro-algae of the silique, the schizochytrium limacinum living bodies is directly used as initial bait to be directly fed to the daphnia-shaped larvae in the seawater, through detection, compared with the daphnia-shaped larvae fed in the form of algae powder, the metamorphosis rate, survival rate, MDA, T-AOC, AKP, ACP activity and the like of the daphnia-shaped larvae fed by the living microalgae composition are obviously improved, the direct feeding of the living microalgae composition is beneficial to controlling the cultivation environment, the direct feeding effect of the living bodies almost violates the industry common knowledge, the algae powder is beneficial to blending in the field of prawn feeding, the nutrition is more sufficient in unit weight, the direct feeding of the algae powder in general cognition is necessarily better than that of the feeding of the living bodies, but the result of the experiment is beneficial to correct bias.
Furthermore, the salinity of the seawater is 25-32, and the density of the daphnia larvae of the litopenaeus vannamei in the seawater is 0.2-0.5 tail/cm3E.g. of sea waterThe salinity is 25, 28, 30, 32, etc., and preferably, the seawater is filtered by a microporous filter membrane, for example, the filtering diameter of the microporous filter membrane is 0.45 mu m, for the density of the daphnia-shaped larvae of the litopenaeus vannamei in the seawater, for example, 5L of seawater is added into a 20L cultivation box, and 1 million daphnia-shaped larvae are added for cultivation, and the density can be properly adjusted within a limited daphnia-shaped larva density range.
In the third aspect of the present application, the application of the living microalgae composition in artemia breeding is disclosed, after artemia hatching, the artemia nauplii are placed in a culture container containing seawater, the density is 800-5cells/ml。
According to the scheme, the living microalgae composition is directly used for cultivating the artemia, the growth of the artemia, the DHA content in the artemia and the like are greatly improved through detection, the nutrition is enhanced, and the method is suitable for being used as bait for cultivating the litopenaeus vannamei or other species such as fish and shrimps.
Furthermore, the salinity of seawater in the culture container is 20-30%, such as 20%, 22%, 25%, 28%, etc.
The fourth aspect of the application document discloses an application of the living microalgae composition in raising the litopenaeus vannamei, wherein the raised artemia are continuously fed to the litopenaeus vannamei, each L of seawater in a culture box contains 2-10 litopenaeus vannamei, the feeding amount of the artemia is 700-.
According to the scheme, the artemia subjected to nutrition enhancement of the living microalgae composition is used as bait and the basic feed is used for cultivating the litopenaeus vannamei, after cultivation is finished, the body length and the growth rate of the litopenaeus vannamei, the total amount of amino acid, the total amount of essential amino acid, the total amount of umami amino acid, DHA and the like in muscle of the litopenaeus vannamei are greatly improved, and the comprehensive value of the cultivated litopenaeus vanname.
Furthermore, the water temperature in the culture box is 22-28 ℃, the salinity is 15-21 per thousand, the dissolved oxygen is more than 5mg/L, the pH is 7.0-8.5, if the water temperature is 28 ℃, the salinity is 18 per thousand, the pH is about 7.5, the dissolved oxygen is 6mg/L, or the warm water temperature is 25 ℃, the salinity is 20 per thousand, the pH is about 7.3, the dissolved oxygen is 7mg/L, and the like, and the temperature can be freely adjusted in the range.
Further, the ammonia nitrogen in the culture box is less than 0.50mg/L, and the nitrite is less than 0.15 mg/L.
Further, the feeding amount of the basic feed is equal to the quality of the artemia, the basic feed is fed for the same times, for example, the basic feed is fed for 4 times every day, and the basic feed is fed for the same quality after the artemia are fed.
Further, the basic feed components include water, crude protein, crude fat, and crude ash, although other components may be added to the above.
Compared with the prior art, the invention has the beneficial effects that:
1. when the living microalgae composition is applied to the daphnia hyaline larvae, the metamorphosis rate survival rate, MDA, T-AOC, AKP, ACP activity and the like of the daphnia hyaline larvae can be greatly improved.
2. When the living microalgae composition is applied to artemia cultivation, the growth of the artemia and the DHA content in the body can be greatly improved, and the nutritional components of the artemia are enhanced.
3. When the living microalgae composition is applied to the cultivation of litopenaeus vannamei, the body length growth rate and the specific growth rate of the litopenaeus vannamei, the total amount of amino acid, essential amino acid, the total amount of umami amino acid, DHA and the like in the muscle of the litopenaeus vannamei can be greatly improved, and the comprehensive value of the cultivated litopenaeus vannamei is high.
Drawings
FIG. 1 is a graph of the effect of different microalgae on survival of litopenaeus vannamei larvae;
FIG. 2 is a graph showing the effect of microalgae on the activity of the immune enzyme of daphnia larvae of litopenaeus vannamei;
FIG. 3 is a graph of the effect of different microalgae on artemia growth;
FIG. 4 is a graph of the effect of microalgal nutrient enrichment of artemia on the growth of litopenaeus vannamei;
FIG. 5 is a graph of the effect of microalgal nutrient enrichment artemia on litopenaeus vannamei digestive enzymes;
FIG. 6 is a graph of the effect of living schizochytrium limacinum and algae meal baits on artemia growth;
in the figure: control represents an uninforced artemia group, SL represents a schizochytrium limacinum nutrition-enhanced artemia group, IG represents an isoflagellate nutrition-enhanced artemia group, CG represents a chaetoceros gracilis nutrition-enhanced artemia group, CV represents a chlorella nutrition-enhanced artemia group, IG + SL represents isoflagellate + schizochytrium limacinum nutrition-enhanced artemia group, CG + SL represents a chaetoceros gracilis + schizochytrium limacinum nutrition-enhanced artemia group, and CV + SL represents chlorella + schizochytrium limacinum nutrition-enhanced artemia group.
Detailed Description
The present invention will be further described with reference to the following specific examples.
Schizochytrium incarnatum ATCC 1381(Schizochytrium incarnum ATCC 1381).
Chaetoceros gracilis is supplied by Ningbo Katsumada Biotechnology Ltd.
Litopenaeus vannamei larvae was supplied by guangdong haixing agriculture group ltd.
Artemia (Artemia sp.) are bought from imperial crown brand brine shrimp eggs, produced in *** plateau.
Post-litopenaeus vannamei (Litopenaeus vannamei) was provided by Haxingnong group of agriculture, Inc., Guangdong.
First, culture of microalgae
1.1, f/2 culture medium was sterilized by filtration through a fibrous filter having a pore size of 0.22. mu.m, and the formulation of the f/2 culture medium is shown in the following table
TABLE 1 culture Medium formulation
Schizochytrium limacinum fermentation medium: glucose 50 g.L-15 g.L yeast extract-1Sodium glutamate 4 g.L-125 per mill of natural seawater, and the pH value is 6.8.
1.2 cultivation method of Chaetoceros gracilis and Chaetoceros gracilis
1.21, filtering the fermentation medium of schizochytrium limacinum with a 0.45-micron microporous membrane, sterilizing at 121 ℃ and 15psi for 30min, and cooling to room temperature. Placing the inoculated mixture in a full-temperature oscillation incubator for dark culture at the temperature of 25 ℃ and the rotating speed of a shaking table of 170rpm min-1. Conical flasks used for culturing algae cells are soaked in dilute acid for 24h, washed and dried.
The schizochytrium limacinum was centrifuged at 8000rpm at 4 ℃ for 5min and the supernatant was discarded. Washing algae cells with distilled water, centrifuging under corresponding conditions, removing supernatant, repeating the operation for 3 times, placing the obtained algae mud into a freeze dryer, and freeze drying to constant weight. And (4) storing the collected algae cells at-20 ℃ for later use.
1.22 the Chaetoceros gracilis is prepared by settling seawater (pH 7.8 + -0.1, salinity 28), filtering with 0.45 μm microporous membrane, sterilizing at 121 deg.C and 15psi for 30min, and cooling to room temperature. Preparing algae culture solution with f/2 improved formula under aseptic condition, inoculating, and culturing in MGC-BPY-2 type illumination incubator at 25 deg.C and illumination intensity of 100 μmol-2s-1And D is 12:12 in light dark cycle.
Chaetoceros gracilis was centrifuged at 11000rpm at 4 ℃ for 10min and the supernatant was discarded. Washing algae cells with distilled water, centrifuging under corresponding conditions, removing supernatant, repeating the operation for 3 times, placing the obtained algae mud into a freeze dryer, and freeze drying to constant weight. And (4) storing the collected algae cells at-20 ℃ for later use.
Secondly, culturing daphnia larvae of the litopenaeus vannamei by using living microalgae composition
2.1 Selective culture of daphnia larvae
The cultivation of the litopenaeus vannamei larvae, in a 20L cultivation box (41 multiplied by 29 multiplied by 23.5cm), 5L seawater with the salinity of 30 filtered by a 0.45 mu m microfiltration membrane is respectively added into each cultivation box, 3 test groups are arranged, each group is divided into three groups, and 1 ten thousand daphnia-shaped larvae with good activity and synchronous metamorphosis to the first stage are respectively randomly placed into each test group for cultivation. During the cultivation process, the water body is continuously aerated, and the dissolved oxygen is more than 6 mg.L-1The temperature of the cultivation water is 28 +/-1 ℃.
2.2 feeding of Living microalgae compositions
The test groups are 7 groups, which are respectively: schizochytrium Limacinum (SL), Isochrysis Galbana (IG), Chaetoceros Gracilis (CG), Chlorella (CV), isochrysis galbana + schizochytrium limacinum (IG + SL), chaetoceros gracilis + schizochytrium limacinum (CG + SL), chlorella + schizochytrium limacinum (CV + SL), with three in each group. Each test group was fed with single microalgae or mixed algae solution.
When the larva begins to change into a daphnoid I phase, correspondingly feeding living microalgae compositions in the chaetoceros gracilis and chaetoceros gracilis test group, wherein the mass of the living microalgae compositions is equal to 1:1, feeding the rest test groups with microalgae corresponding to the test group name respectively, and feeding the rest test groups for 4 times every day, wherein the feeding amounts of the chaetoceros gracilis, the flagellates such as globulina, the chaetoceros gracilis and the chlorella are respectively: 1X 104cells·ml-1、3.5×104cells·ml-1、4×104cells·ml-1、1.5×105cells·ml-1The mixed algae was fed according to 1/2 for individual algae. And observing the growth and ingestion of the larva, and checking the content of microalgae in the water to increase or decrease the content of microalgae in the water. The indexes of pH value, dissolved oxygen, temperature, salinity and the like in the aquaculture water body are measured regularly every day.
2.3 determination of larval growth Performance indicators
During the seedling raising period, a certain number of larvae are randomly selected at 16 points every day for microscopic examination, the survival rate and the metamorphosis rate of the larvae are recorded, and the growth condition is observed.
The cultivation period is five days, and statistics shows that the survival rate of the larvae fed by different microalgae shows a decreasing trend along with the increase of the cultivation time, as shown in figure 1.
Finally, the combination of Chaetoceros gracilis and Chaetoceros gracilis is obviously higher than that of the other two groups, the survival rate is 30.22%, and the survival rate of the other groups is between 12 and 15%; the metamorphosis process of daphnia larva is daphnia I phase (Z)1) Daphnia stage II (Z)2) Daphnia III (Z)3) The effect on the metamorphosis rate is shown in the following table
TABLE 2 influence of different microalgae on metamorphosis rate of daphnia-shaped larvae of Litopenaeus vannamei at different stages
Note: different letters in the same column indicate significant difference (P < 0.05).
It can be seen that the number of larvae of the prawn from Z2To Z3During the development process, the schizochytrium limacinum, Chaetoceros gracilis and mixed algae group are gradually shown to be more beneficial to the metamorphosis of prawn larvae, and Z is the group that chlorella cannot support the larvae2Develop to Z3Is more beneficial to the growth and metamorphosis of the larva.
And detecting the influence of microalgae on the activity of the superoxide dismutase (SOD), the total antioxidant capacity T-AOC, the MDA (malonaldehyde) content, the AKP (alkaline phosphatase) activity and the ACP (acid phosphatase) activity of the Litopenaeus vannamei Boone, and determining the specific operation steps of the kit for establishing the bioengineering institute by Nanjing, as shown in figure 2.
It can be seen that the T-AOC, AKP and ACP activities of the Chaetoceros gracilis and the schizochytrium mixed fed to the daphnia larvae of the prawns are significantly higher than those of other groups.
Application of living microalgae composition in cultivation of artemia
3.1 incubation of artemia
The artemia cysts are incubated in a 50L culture box, natural seawater with the salinity of 25 per mill is added, the culture box is placed at the room temperature of 25 ℃ for continuous aeration incubation for 24 hours, and artemia nauplii are separated for experiments.
3.2 enhanced culture of artemia by the live microalgae composition
After the artemia are incubated for 48 hours, selecting the artemia nauplii with good activity for carrying out nutrition enrichment experiment, culturing the artemia in a 5L beaker with the density of 1000 ind.L-1The temperature is 25 ℃, and the salinity is 25 per mill.
In the experiment, 7 kinds of microalgae are treated, and 3 parallel groups are respectively arranged without feeding the microalgae as control groups. The treatment of 7 groups of microalgae respectively comprises: the densities of the cells of the schizochytrium limacinum, the Isochrysis Galbana (IG), the Chaetoceros Gracilis (CG), the Chlorella (CV), the isochrysis galbana + Schizochytrium limacinum (IG + SL), the Chaetoceros gracilis + Schizochytrium limacinum (CG + SL), and the chlorella + Schizochytrium limacinum (CV + SL) fed by the parachryma, the isochrymala, the Chaetoceros gracilis and the chlorella are respectively as follows according to the biomass and the size of the microalgae: 1X 105cells·ml-1、3.5×105cells·ml-1、4×105cells·ml-1、1.5×106cells·ml-1The mixed algae was fed according to 1/2 for individual algae.
After the nutrition is enhanced for 12h, artemia larvae are collected, algae cells remained on the larvae are washed for many times by sterile water, and the larvae are stored in a freezing storage tube after being washed and are stored at the temperature of minus 80 ℃ to be tested.
Meanwhile, 4 groups of test groups were set, and cultured with Schizochytrium Sp (SL), Isochrysis Galbana (IG), Chaetoceros Gracilis (CG) and Chlorella (CV), respectively, and the feeding density was 1 × 104cells·ml-1、3.5×104cells·ml-1、4×104cells·ml-1、1.5×105cells·ml-1The density of the artemia is 500ind & L-1Culturing in 1L beaker at 25 deg.C and salinity of 25 ‰. The culture period is one week, aeration is carried out in the culture process, water is changed by 50% every other day, feeding is carried out for three times every day, the body length of the artemia is measured, and the four groups are normal culture to form contrast with intensified culture.
In addition, the algae powder and living body bait feeding test groups are respectively a schizochytrium limacinum living body feeding group, a schizochytrium limacinum powder feeding group and a control group without bait, wherein the algae powder and the schizochytrium limacinum living body are fed for the same times with equal mass and the like, the environment is consistent, as can be seen from figure 6, artemia hardly grow in the control group (without bait feeding) and almost die in the fourth day, the living schizochytrium limacinum is used for feeding the artemia, the growth of the artemia is obviously higher than that of the schizochytrium limacinum powder feeding group, and the living schizochytrium limacinum is more favorable for the growth of the artemia.
3.3, detection
3.31 Effect of different microalgae on artemia growth
After intensive culture and normal culture, the test groups fed with the chaetoceros gracilis (SL), the Isochrysis Galbana (IG), the Chaetoceros Gracilis (CG) and the Chlorella Vulgaris (CV) have the greatest increase in the body length of the artemia from 0.8-0.9mm to about 2.4mm, the chaetoceros gracilis group from 0.8-0.9mm to about 1.5mm, and the artemia body length of the chaetoceros gracilis and the chaetoceros gracilis group from 0.8-0.9mm to about 2.0mm, as shown in FIG. 3.
After the enhanced culture, fatty acid detection is shown in Table 3
TABLE 3 Effect of microalgal Nutrition fortification on artemia fatty acid composition
The above table shows that the content of DHA + EPA in a single artemia in the schizochytrium limacinum group accounts for 2.6517 +/-0.1570% of the total fatty acid content, the content of DHA/EPA is 0.1364 +/-0.0045, the content of DHA + EPA in a single artemia in the chaetoceros gracilis group accounts for 2.0332 +/-0.0191 of the total fatty acid content, the content of DHA/EPA is 0.0255 +/-0.0009, the content of DHA + EPA in a single artemia in the living microalgae combination group accounts for 2.1119 +/-0.0428 of the total fatty acid content, but the chaetoceros gracilis and the schizochytrium limacinum in the living microalgae combination group can be selected to be in a better state according to the needs, the total mass of DHA is increased under the combination, and the value is improved.
Application of living microalgae composition in litopenaeus vannamei
4.1, setting 8 groups of tests, respectively feeding dinoflagellate chrysosporium (SL), Isochrysis Galbana (IG), Chaetoceros Gracilis (CG), Chlorella (CV), isochrysis galbana + schizochytrium (IG + SL), chaetoceros gracilis + schizochytrium (CG + SL) and chlorella + schizochytrium (CV + SL) reinforced artemia, feeding non-reinforced artemia to a control group, repeating the steps by 3 groups, placing 50 postlarvae shrimp larvae in each breeding box, and after feeding the artemia each time, properly feeding a basic shrimp feed with the same quality and the constant source brand on the market as an auxiliary feed to each experimental group, wherein the components of the basic feed are shown in tables 4 and 5.
Before the experiment, P is firstly added15Temporarily culturing the prawns in a 50L culture box for one week, and feeding a constant source shrimp larva basic feed during the temporary culture. After the temporary culture is finished, healthy and active shrimp fries with consistent specifications are selected to a culture box (41 multiplied by 29 multiplied by 23.5cm) containing 10L seawater for formal test. The initial weight of the young litopenaeus vannamei is 0.05 plus or minus 001g (mean. + -. SD), initial body length 1.14. + -. 0.15cm (mean. + -. SD).
During the experiment, feeding is carried out for 4 times every day at regular time, the initial feeding amount of the artemia is 1000ind per group, the auxiliary feed is 2% of the initial weight of the prawns, the feeding amount is adjusted in time every day according to the ingestion condition, redundant feed and excrement are removed, 20% of seawater is replaced, all seawater is replaced in 5-7 days, and the seawater used in the experiment is settled and filtered by a 0.45-micrometer microporous filter membrane. The culture period is 6 weeks, the water quality is measured regularly during the test period, the water temperature is 28 + -1 deg.C, the salinity is 18 ‰, and the dissolved oxygen is more than 5 mg.L-1pH of 7.8-8.2, nitrite content less than 0.15 mg.L-1The ammonia nitrogen is less than 0.50 mg.L-1。
TABLE 4 basic Nutrition ingredients of feed
TABLE 5 composition and content of amino acids and fatty acids in feed
And 4.2, detecting various performances of the cultured shrimps in each test group.
4.21 sample Collection
After the 6-week experiment period, all prawns in each group of experiments were counted, weighed, and weighed. 10 shrimps were randomly selected from each experimental group and dissected under ice bath conditions, surgical scissors and forceps for dissection were sterilized, liver, pancreas and intestinal tract were separated after dissection, and after washing with sterile physiological saline, the shrimps were placed in a 2ml cryopreservation tube and stored at-80 ℃ for measurement of related enzyme activities.
4.22, the influence of the microalgae nutrition-enriched artemia on the muscle nutrient composition of the litopenaeus vannamei is shown in the following table and the figure.
TABLE 6 influence of microalgae nutrition-enriched artemia on the muscle amino acid composition of Litopenaeus vannamei
Note: different letters on the same row indicate significant difference (P < 0.05). The ones marked with x are umami amino acids.
It can be seen that the total amount of muscle amino acids of the microalgae nutrition-enhanced artemia cultured litopenaeus vannamei is as follows: chaetoceros gracilis group (837.29mg g)-1)>Isochrysis galbana schizochytrium group (825.26mg g)-1)>Isochrysis galbana (809.14mg g)-1)>Artemia group (802.23mg g) without nutrient enrichment-1)>Chaetoceros gracilis schizochytrium group (800.63mg g)-1)>Chlorella group (795.69mg g)-1)>Schizochytrium limacinum group (795.1mg g)-1)>Chlorella and Chlamydomonas split group (750.96mg g)-1). The total amount of amino acid, the total amount of essential amino acid, the total amount of semi-essential amino acid, the total amount of nonessential amino acid and the total amount of delicious amino acid in the muscle of the litopenaeus vannamei cultivated by the dinoflagellate group, the dinoflagellate group and the schizochytrium sp nutrition-enhanced artemia are obviously higher than those of other experimental groups.
TABLE 7 influence of microalgae nutrition-enriched artemia on the fatty acid composition of Litopenaeus vannamei
Note: different letters in the same row indicate significant differences (P < 0.05). The symbol "- -" indicates no detection.
It can be seen that the control group and schizochytrium limacinum group detected C22:1 and C22:5, while the other experimental groups did not. The muscle fatty acid composition and content of different microalgae nutrition-enhanced artemia fed to litopenaeus vannamei are different. The total 15 kinds of fatty acid detected in the prawn muscle are higher in content of C16:0 and C20: 5. The total fatty acid content, the saturated fatty acid content and the polyunsaturated fatty acid content of each experimental group are all higher than those of a control group, and the content of C20:4 in the control group is also lower than those of other experimental groups. The content of EPA and DHA in the muscles of the prawns is different, the content of DHA in the muscles of the schizochytrium limacinum group and the prawns mixed with the schizochytrium limacinum is obviously higher than that of DHA in the muscles of other experimental groups (P <0.05), and the content of DHA in the muscles of the prawns and the muscles of the prawns mixed with the schizochytrium limacinum is more than 6% of the total fatty acid content, and is increased by 1..
4.3, determination of influencing factors of the weight, body length and growth of the prawns
Wherein the determination of body weight and body length is: at the end of the experiment, all prawns from each group were subjected to body weight and length measurements. Absorbing water on the body surface of the prawns by using absorbent paper, weighing by using balance, and recording the body weight (g). The total length of the prawns, i.e. the length from the front end of the frontal angle to the end of the tail section of the prawns, was measured using a ruler and the body length (cm) was recorded.
The influence of the microalgae nutrition-enhanced artemia on the growth of litopenaeus vannamei is shown in fig. 4, wherein the specific calculation method is as follows: survival rate (Survival rate,%) 100 × (final prawn tail)/(initial prawn tail)
Weight gain ratio (WGR,%) 100 × (final average Weight-initial average Weight)/initial average Weight
Growth rate (GR,%) 100 × (final body length-initial body length)/initial body length Specific Growth rate (Specific Growth rate, SGR% >, day)-1) 100 × (ln (final average weight) -ln (initial average weight))/days of experiment.
It can be seen that, after the feeding experiment of 6 weeks, the survival rates of the nutrition-enhanced artemia fed to the prawns by the dinoflagellate group such as the chlorella and the chlorella + schizochytrium group (fig. 4A) are 65.33% and 63.33% respectively, which are significantly higher than those of other experimental groups (p <0.05), the survival rate of the chlorella group is lower and is 42.67%, and the difference between the other experimental groups is not significant (p > 0.05). In addition, the body length (fig. 4E), the terminal average weight (fig. 4F), the weight gain rate (fig. 4D), the growth rate (fig. 4C) and the specific growth rate (fig. 4B) of the schizochytrium limacinum group of the shrimp are all significantly higher than those of other groups (p <0.05), and are respectively improved by 18.17%, 28.87%, 30.13%, 11.68% and 7.30% compared with the control group, and the influence of each experimental group on the body length and the growth rate of the litopenaeus vannamei is mainly shown as follows: schizochytrium limacinum, chaetoceros gracilis and control group.
And the influence of the microalgae nutrition-enhanced artemia on intestinal digestive enzyme of the litopenaeus vannamei is detected, as shown in fig. 5, it can be seen that the chymotrypsin activity of the Chaetoceros gracilis and schizochytrium limacinum-enhanced artemia group is obviously higher than that of other experimental groups, namely digestion is promoted, absorption is enhanced, and the body length, the growth rate and the like are greatly improved; the effects on the weight average, the weight gain rate and the specific growth rate are: schizochytrium sp.sp.gracilis > control group > chaetoceros gracilis plus schizochytrium gracilis > chaetoceros gracilis.
For review, the combination of Chaetoceros gracilis and Chaetoceros gracilis greatly improves the body length, the growth rate, the muscle amino acid content, the DHA and the like of the prawns, and the comprehensive value of the cultured prawns is high.
The above is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above-mentioned embodiments, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the invention may occur to those skilled in the art without departing from the principle of the invention, and are considered to be within the scope of the invention.
Claims (10)
1. The living microalgae composition is used for feeding artemia, daphnia larvae of litopenaeus vannamei and shrimps, and is characterized in that: comprises the following components: fine horn hair microalgae, schizochytrium limacinum.
2. The application of the living microalgae composition in the feeding of the daphnia larvae of the litopenaeus vannamei is characterized in that: culturing daphnia larva of Litopenaeus vannamei in a culture tank containing seawater, maintaining seawater dissolved oxygen amount at more than 6mg/L and water temperature at 25-30 deg.C, feeding micro-algae and schizochytrium living body composition when larva begins to become daphnia, and feeding at density of 2-4 × 104cells/ml。
3. Use of the live microalgae composition according to claim 2 for feeding the daphnia larvae of litopenaeus vannamei: the salinity of seawater is 25-32, and the density of the daphnia-shaped larva of Litopenaeus vannamei in seawater is 0.2-0.5 tail/cm3。
4. The application of the living microalgae composition in the feeding of artemia is characterized in that: after the artemia are hatched, the artemia nauplii are placed in a culture container containing seawater, the density is 800-5cells/ml。
5. Use of a live microalgae composition according to claim 4 in artemia feeding, wherein: the salinity of the seawater in the culture container is 20-30 per mill.
6. The application of the living microalgae composition in the breeding of the litopenaeus vannamei is characterized in that: the artemia bred in the claim 4 are continuously fed to the litopenaeus vannamei, 2-10 litopenaeus vannamei are contained in each L of seawater in the breeding box, the feeding amount of the artemia is 700-.
7. Use of the live microalgae composition of claim 6 in the breeding of post-litopenaeus vannamei, characterized in that: the water temperature in the culture box is 22-28 ℃, the salinity is 15-21 per mill, the dissolved oxygen is more than 5mg/L, and the pH is 7.0-8.5.
8. Use of the live microalgae composition of claim 6 in the breeding of post-litopenaeus vannamei, characterized in that: the ammonia nitrogen in the culture box is less than 0.50mg/L, and the nitrite is less than 0.15 mg/L.
9. Use of the live microalgae composition of claim 6 in the breeding of post-litopenaeus vannamei, characterized in that: the feeding amount of the basic feed is equal to the quality of the artemia, and the basic feed is fed for the same times.
10. Use of the live microalgae composition of claim 9 in the breeding of post-litopenaeus vannamei, characterized in that: the basic feed components comprise water, crude protein, crude fat and crude ash.
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