CN112425537A - Method for improving survival rate and attachment rate of young apostichopus japonicas by adding alpha-amylase and application of method - Google Patents
Method for improving survival rate and attachment rate of young apostichopus japonicas by adding alpha-amylase and application of method Download PDFInfo
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- 102000004139 alpha-Amylases Human genes 0.000 title claims abstract description 46
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- 238000000034 method Methods 0.000 title claims abstract description 21
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- 241000965254 Apostichopus japonicus Species 0.000 claims description 22
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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
- A01K61/30—Culture of aquatic animals of sponges, sea urchins or sea cucumbers
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/189—Enzymes
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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 survival rate and attachment rate of young stichopus japonicas by adding alpha-amylase and application thereof. The method is characterized in that a certain dose of alpha-amylase is added into young stichopus japonicas larva bait from the initial stage of development of young stichopus japonicas into large ear larva, compared with a control group without alpha-amylase, the survival rate of the planktonic larvae of the young stichopus japonicas in the test group is improved by 24.2%, the attachment rate of the larvae of the young stichopus japonicas is improved by 4.3 times, the survival rate and the attachment rate of the larvae of the small stichopus japonicas are obviously improved, and therefore the bottleneck problem of high larva death rate in the artificial breeding process of the young stichopus japonicas is solved.
Description
Technical Field
The invention belongs to the technical field of mariculture biology, and particularly relates to a method for improving survival rate and attachment rate of young apostichopus japonicas by adding alpha-amylase and application thereof.
Background
The Xiaoverruca stichopus is commonly called as 'yellow meat ginseng', and becomes a representative high-economic-value variety in tropical sea cucumbers in China due to high nutritional value and good mouthfeel.
The artificial breeding of the stichopus japonicas is initially successful in recent years, but the death rate of the stichopus japonicas in the larval development period is high, and the stichopus japonicas becomes a bottleneck of the artificial breeding of the stichopus japonicas. The development of the young apostichopus japonicus is mainly carried out in the development stages of blastocyst, gastral embryo, ear larva (small ear larva and big ear larva), goblet-shaped larva, five tentacle larva, young stichopus japonicus and the like. Our previous studies showed that the mortality rate of young apostichopus japonicus from big ear larvae (planktonic larvae) to goblet-shaped larvae (planktonic larvae) and from goblet-shaped larvae to pentahapto larvae (adherent larvae) is extremely high, with only 2-3% of planktonic larvae successfully metamorphosing to pentahapto larvae and successfully adhering. Therefore, how to improve the survival rate and the attachment rate of the young stichopus japonicas becomes a key problem which needs to be solved urgently by the artificial breeding of the stichopus japonicas.
Alpha-amylase is one of the most common extracellular enzymes that hydrolyze starch and glycogen molecules. The main hydrolysis mode of the enzyme is the disruption of the alpha-1, 4-glucosidic bonds on long chains of starch, producing glucose, maltose, maltotriose and alpha-limiting dextrins. There is currently no relevant research on the use of alpha-amylase to improve survival and attachment rates of young apostichopus japonicas.
Disclosure of Invention
The first purpose of the invention is to provide a method for improving the survival rate and the attachment rate of young stichopus japonicas, which is to add a certain dose of alpha-amylase to the bait of young stichopus japonicas from the initial stage of the young stichopus japonicas developing into big ear young stichopus japonicas so as to improve the survival rate and the attachment rate of the young stichopus japonicas, thereby solving the bottleneck problem of high death rate of young stichopus japonicas in the artificial breeding process of the young stichopus japonicas.
Experiments prove that the survival rates of the parvalonia verrucosa planktonic larvae (69.2 percent and 61.86 percent respectively) added with the alpha-amylase with the final concentrations of 20U/L and 10U/L are respectively improved by 24.2 percent and 16.86 percent compared with a control group (the survival rate is 45 percent) not added with the alpha-amylase.
The adhesion rate (13.14%) of the young stichopus parviflorus added with the alpha-amylase with the final concentration of 10U/L is remarkably higher than that of a control group (the adhesion rate is 2.48%) without the alpha-amylase and an experimental group added with the alpha-amylase with other concentrations, and the adhesion rate is improved by 4.3 times compared with that of the control group.
Therefore, the final concentration of the alpha-amylase is preferably 5U/L to 20U/L.
Preferably, the method for improving the survival rate and the attachment rate of the young apostichopus japonicas is characterized in that alpha-amylase with the final concentration of 5-20U/L is added into the culture water body in the first 10 days from the initial stage of the young apostichopus japonicas developing into the young auricle, and the alpha-amylase with the final concentration of 5-10U/L is added in the last 10 days to improve the survival rate and the attachment rate of the young apostichopus japonicas.
More preferably, the method for improving the survival rate and the attachment rate of the young apostichopus japonicas is to add alpha-amylase with the final concentration of 20U/L into the aquaculture water body 10 days before the young apostichopus japonicas develops into the young auricle, and add the alpha-amylase with the final concentration of 10U/L into the aquaculture water body 10 days after the young apostichopus japonicas develops into the young auricle, so that the survival rate and the attachment rate of the young apostichopus japonicas are improved.
Preferably, the bait comprises algae powder, yeast and young sea cucumber feed.
Preferably, the algae powder comprises chlorella powder, sargassum thunbergii powder and sargassum powder.
The second purpose of the invention is to provide the application of alpha-amylase in preparing the preparation for improving the survival rate and the attachment rate of young stichopus japonicas.
The third purpose of the invention is to provide a preparation for improving the survival rate and the attachment rate of young stichopus japonicas, which contains alpha-amylase.
The invention has the following beneficial effects:
the invention provides a method for improving survival rate and attachment rate of young stichopus japonicas by adding alpha-amylase. The method is characterized in that a certain dose of alpha-amylase is added into young stichopus japonicas bait from the initial stage of development of young stichopus japonicas into large ear larvae, compared with a control group without alpha-amylase, the survival rate of planktonic larvae of the young stichopus japonicas is improved by 24.2 percent, the attachment rate of the larvae of the young stichopus japonicas is improved by 4.3 times, the survival rate and the attachment rate of the larvae of the small stichopus japonicas are obviously improved, and therefore the bottleneck problem of high larva death rate in the artificial breeding process of the young stichopus japonicas is solved.
Drawings
FIG. 1 is a graph showing the effect of various concentrations of alpha-amylase on survival of planktonic larvae of Acanthopanax parviflorus. Note: the different lower case letters between experimental groups in the figure indicate significant difference (P <0.05) and the same letter indicates no significant difference.
FIG. 2 is a graph showing the effect of various concentrations of alpha-amylase on the rate of adhesion of juveniles of Apostichopus parviflora. Note: the different lower case letters between experimental groups in the figure indicate significant difference (P <0.05) and the same letter indicates no significant difference.
Detailed Description
In order to make the objects, technical solutions and advantageous technical effects of the present invention clearer, the present invention is further described in detail with reference to the following embodiments. It should be understood that the embodiments described in this specification are only for the purpose of illustrating the invention and are not to be construed as limiting the invention, and the parameters, proportions and the like of the embodiments may be suitably selected without materially affecting the results. The examples are all reagents and process steps conventional in the art, except where specifically indicated. Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
Example 1
1. Effect of different concentrations of alpha-amylase on survival rate of young stichopus parviflorus
(1) Collection and transfer of young Proteus parviflorus
Observing young small stichopus japonicus in the culture pond by using an optical microscope, collecting young small stichopus japonicus in the large ear period when 80% of the young small stichopus japonicus in the culture pond is in the large ear period, and randomly transferring the young small stichopus japonicus to 6 large white buckets filled with filtered seawater with 100L and the salinity of 35 per thousand, so that the density of the large ear young in each bucket is 100 per L.
(2) alpha-Amylase addition experiments
5 days before the experiment, 0.05g of chlorella powder and 0.05g of yeast were fed to each bucket every day, and alpha-amylase was added to each of buckets No. 2-6 to final concentrations of 5U/L, 10U/L, 20U/L, 50U/L and 100U/L, respectively. From day 6, 0.05g of chlorella powder, yeast, sargassum powder, gulfweed powder and young sea cucumber feed (Dalian flood dragon aquatic feed development center) are fed into each barrel every day, and alpha-amylase is added into barrels 2 to 6 respectively until the final concentrations are 5U/L, 10U/L, 20U/L, 50U/L and 100U/L.
(3) Statistics of survival rate of young stichopus parviflorus
On the 10 th day of the experiment, 100mL of water was randomly taken from a large white bucket with a beaker, and the number of planktonic larvae in the beaker was counted visually 5 times per bucket. The planktonic larvae survival rate in each bucket was calculated by dividing the current planktonic larvae density by the initial planktonic larvae density. As shown in FIG. 1, the survival rate of planktonic larvae of stichopus parviflorus was the highest (69.2%) in the experimental group added with alpha-amylase at a final concentration of 20U/L, and was increased by 24.2% and 16.86% in the experimental group added with alpha-amylase at a final concentration of 10U/L (61.86%) compared to the control group not added with alpha-amylase (45%).
2. Effect of different concentrations of alpha-Amylase on the adhesion rate of young Apostichopus parvifolius
The counted young stichopus japonicus is put back into the barrel for continuous feeding, 0.05g of chlorella powder, yeast, sargassum powder, gulfweed powder and young stichopus japonicus feed (Dalian dragon aquatic feed development center) are continuously fed into each barrel every day, and alpha-amylase is respectively added into No. 2-6 barrels until the final concentration is 5U/L, 10U/L, 20U/L, 50U/L and 100U/L. On the 20 th day of the experiment, when all the planktonic larvae in the vat were transformed into adherent larvae, all the small stichopus japonicus adherent larvae in the vat were collected, all the larvae were transferred to 100mL of water, after the mixture was blown up uniformly, 1mL of the mixture was taken out, the number of adherent larvae was observed and counted under a microscope, and the procedure was repeated 5 times. The attachment rate of young scoliosis parva per large white bucket was calculated by dividing the total number of attached larvae by the number of initial planktonic larvae. As shown in FIG. 2, the adhesion rate of the juveniles of the stichopus parviflorus in the experimental group added with the alpha-amylase at the final concentration of 10U/L is the highest (13.14%), which is significantly higher than that of the control group (the adhesion rate is 2.48%) (P is less than 0.001) added with the alpha-amylase at other concentrations, and the adhesion rate is improved by 4.3 times compared with that of the control group. Therefore, the alpha-amylase with the final concentration of 10U/L can be added into the culture water body during the young stichopus japonicas to improve the survival rate and the attachment rate of the young stichopus japonicas.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.
Claims (6)
1. A method for improving the survival rate and the attachment rate of young stichopus japonicus juveniles is characterized in that alpha-amylase is added into bait from the initial stage of the young stichopus japonicus juveniles developing into big ear young stichopus japonicus juveniles.
2. The method for improving survival rate and attachment rate of young apostichopus japonicus selenka of claim 1, wherein the alpha-amylase is added to a final concentration of 5-20U/L.
3. The method for improving the survival rate and the attachment rate of the young apostichopus japonicus selenka as claimed in claim 1 or 2, wherein the alpha-amylase with the final concentration of 5-20U/L is added into the aquaculture water body in the first 10 days from the initial stage of the young apostichopus japonicus selenka developing into the young auricle, and the alpha-amylase with the final concentration of 5-10U/L is added in the last 10 days to improve the survival rate and the attachment rate of the young apostichopus japonicus selenka.
4. The method for improving the survival rate and the attachment rate of the young apostichopus japonicus selenka as claimed in claim 3, wherein the alpha-amylase with the final concentration of 20U/L is added into the aquaculture water body 10 days before the young apostichopus japonicus selenka develops into the young auricle, and the alpha-amylase with the final concentration of 10U/L is added in the next 10 days to improve the survival rate and the attachment rate of the young apostichopus japonicus selenka.
5. Application of alpha-amylase in preparation of a preparation for improving survival rate and attachment rate of young apostichopus japonicus juveniles.
6. A preparation for improving the survival rate and the attachment rate of young apostichopus japonicus juveniles is characterized by comprising alpha-amylase.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008011721A (en) * | 2006-07-03 | 2008-01-24 | Tadamasa Fujimura | Method for producing biomass fuel, and method for producing superheated-steam processed product |
| CN101204197A (en) * | 2007-12-06 | 2008-06-25 | 中国水产科学研究院黄海水产研究所 | Ingredients of trepang compound feedstuff additive |
| CN108617558A (en) * | 2018-08-30 | 2018-10-09 | 山东富瀚海洋科技有限公司 | A kind of artificial breeding method of juvenile stichopus |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008011721A (en) * | 2006-07-03 | 2008-01-24 | Tadamasa Fujimura | Method for producing biomass fuel, and method for producing superheated-steam processed product |
| CN101204197A (en) * | 2007-12-06 | 2008-06-25 | 中国水产科学研究院黄海水产研究所 | Ingredients of trepang compound feedstuff additive |
| CN108617558A (en) * | 2018-08-30 | 2018-10-09 | 山东富瀚海洋科技有限公司 | A kind of artificial breeding method of juvenile stichopus |
Non-Patent Citations (3)
| Title |
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| 常青等: "微颗粒饲料中添加消化酶对半滑舌鳎仔、稚鱼生长和消化酶活性的影响", 《海洋水产研究》 * |
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| 武明欣 等: "酶制剂对刺参生长、体成分、免疫能力及氨氮胁迫下免疫酶活力和热休克蛋白70含量的影响", 《动物营养学报》 * |
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