CN111000092A - Enhancer for improving immunity of aquatic animals, preparation method thereof and application thereof in feed - Google Patents
Enhancer for improving immunity of aquatic animals, preparation method thereof and application thereof in feed Download PDFInfo
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- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
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- A23K20/00—Accessory food factors for animal feeding-stuffs
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- A23K20/10—Organic substances
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- A23K20/132—Heterocyclic compounds containing only one nitrogen as hetero atom
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- A23K20/142—Amino acids; Derivatives thereof
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Abstract
The invention discloses an enhancer for improving immunity of aquatic animals, which comprises the following components in mass concentration: 15-20g/L of concentrated molasses fermentation liquor, 4-8g/L of melatonin, 15-20g/L of sodium alginate, 5-8g/L of nitrogen source, 3-5g/L of phosphorus source, 1-2g/L of potassium source, 3-5g/L of high-activity humic acid and 10-15 mu g/L of selenium methionine. Also discloses a preparation method and application of the reinforcing agent. The enhancer has the characteristics of addressing both the symptoms and root causes, safety, no toxicity, no drug resistance, no pollution, no residue and the like, has the effects of bacteriostasis and antibiosis, and can improve the immunity of aquatic animals, resist bacteria, prevent and treat parasites, resist viruses and the like; the enhancer is suitable for aquatic animals in various growth stages, and can especially solve the problem of immunity reduction of cultured animals induced by high temperature season and severe weather, so as to improve survival rate.
Description
Technical Field
The invention belongs to the technical field of aquaculture, and particularly relates to an enhancer for improving immunity of aquatic animals, a preparation method thereof and application thereof in feed.
Background
At present, in an intensive and high-density breeding mode, the problem of exposure in the breeding process caused by the increasingly worsened ecological environment is increasingly prominent, such as disease outbreak, particularly high-temperature seasons, high breeding water temperature, influence of high-temperature stress on bred animals, low immunity, slow growth and the like caused by abnormal ingestion, and the development of aquaculture industry is seriously influenced. The problems of bacterial drug resistance, drug residues and the like caused by the use of a large amount of antibiotics and some prohibited drugs have attracted attention and anxiety of consumers. At present, the method commonly adopted for treating aquatic animal diseases at home and abroad is 'prevention-oriented', and the fundamental purpose is to improve the self-immunity of cultured animals. Vaccination is an effective way to prevent disease in aquaculture, but research on effective vaccines for aquatic animals is slow. The immunopotentiator can improve cellular and humoral immunity and enhance resistance. Research on the development and application of immunopotentiators for aquatic animals has become a research hotspot at present.
The immunopotentiator is a substance which is added into feed or a culture environment, improves the nutritional condition and the environmental condition of aquatic animals by improving the immunity of the aquatic animals, and can enhance the resistance of the aquatic animals to pathogenic microorganisms. The nutrition and the immunity of the aquatic animals have a close synergistic relationship, and the immune function can be adversely affected by unbalanced or insufficient nutrition in the feed. The immunopotentiator has the characteristics of stable performance, wide action, safety, high efficiency and the like, and becomes one of the important ways for solving the problem of green prevention and control of aquatic animal diseases at present. However, the reinforcing agent in the current market cannot well cooperate with feed so as to well improve the immunity of aquatic animals, so that the aquatic animals are influenced by some special conditions, such as high temperature, high density, water quality deterioration and the like, and the problems of appetite reduction, nutrition deficiency, immunity reduction and the like still occur, and the survival rate is low. The search for a special aquaculture animal enhancer synergistic feed to improve the existing problems is a problem which needs to be solved by the technical personnel in the field.
Disclosure of Invention
An object of the present invention is to solve at least the above problems and to provide at least the advantages described later.
The invention aims to provide an enhancer for improving the immunity of aquatic animals, and the formula has the characteristics of addressing both the symptoms and root causes, safety, no toxicity, no drug resistance, no pollution, no residue and the like, has the functions of bacteriostasis and antibiosis, and can obviously reduce the incidence rate of problems of appetite reduction, insufficient nutrition, immunity reduction and the like of the aquatic animals caused by factors such as high temperature, high density, water quality deterioration and the like.
The invention also aims to provide a preparation method of the enhancer for improving the immunity of aquatic animals, which has the advantages of easily obtained raw materials, convenient operation and suitability for mass production and application.
The invention also aims to provide the application of the enhancer for improving the immunity of the aquatic livestock in the basic feed, which is applied to the rapid growth period of different aquaculture varieties, has good stress prevention effect and can obviously solve the problem of the reduction of the immunity of the aquatic livestock induced by high-temperature seasons and severe weather.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides an enhancer for improving immunity of aquatic animals, which comprises the following components in mass concentration: 15-20g/L of concentrated molasses fermentation liquor, 4-8g/L of melatonin, 15-20g/L of sodium alginate, 5-8g/L of nitrogen source, 3-5g/L of phosphorus source, 1-2g/L of potassium source, 3-5g/L of high-activity humic acid and 10-15 mu g/L of selenium methionine.
Preferably, the strain also comprises 2-10g/L of composite EM (purchased from Jinan chemical industry Co., Ltd.).
Preferably, the nitrogen source is urea; the phosphorus source is monoammonium phosphate; the potassium source is potassium hydroxide: the potassium nitrate is mixed according to the mass ratio of 1: 2-3.
Preferably, the high-activity humic acid is prepared by the following method: adding humic acid into an alkali solution according to the weight ratio of 1:6-1:10 for dissolving, heating to 80-100 ℃, reacting for 40-60min under the pressure of 1-2Pa, cooling to room temperature, and continuously hydrolyzing for 5-6h to obtain the high-activity humic acid.
Preferably, the alkali solution is a potassium hydroxide solution, and the potassium hydroxide solution is prepared by mixing potassium hydroxide and water according to the weight ratio of 1:8-1: 10.
The invention also provides a preparation method of the enhancer for improving the immunity of aquatic animals, which comprises the following steps: adding the concentrated molasses fermentation liquor into the high-activity humic acid solution according to the component dosage of the enhancer for improving the immunity of the aquatic animals, adding the nitrogen source, the phosphorus source, the potassium source, the melatonin, the selenium methionine, the sodium alginate and the compound EM bacteria, and uniformly mixing to obtain the enhancer.
The invention also provides application of the enhancer for improving the immunity of the aquatic animals in basic feed, and the enhancer is applied to feed for aquaculture of the aquatic animals.
Preferably, the volume concentration of the enhancer added to the basal feed is 10-20 mL/kg.
Preferably, the reinforcing agent is uniformly sprayed on the surface of the basic feed in a spraying mode and then dried in the shade for 45-60min to obtain the experimental feed. The experimental feed can be stored at a normal-temperature ventilating and dark place for later use after being sprayed and added with the immunopotentiator.
The invention discloses the following technical effects:
1) according to the characteristics of nutrition and immunity requirements of aquaculture animals, a large number of experiments prove that the enhancer special for improving the immunity of the aquaculture animals is obtained, the enhancer provides amino acids, trace elements and the immunopotentiator which are necessary for the aquaculture animals, the growth speed, the anti-stress capability and the immunity capability of the aquaculture animals can be effectively improved in a high-density culture mode, and the enhancer has the advantages of convenience in use, low cost and the like.
2) The preparation method of the enhancer for improving the immunity of the aquatic animals, provided by the invention, has the advantages of abundant and easily-obtained raw material sources and simple operation, does not need professional equipment and professional technicians for preparing the feed, is suitable for wide popularization and application, and has important practical significance and use value.
3) The invention also discloses the application of the reinforcing agent in basic feed, in particular to feed for aquaculture. The aquatic livestock is scientifically bred according to factors such as the growth cycle of the aquatic livestock, the ingestion rhythm of a breeding object and the like, the growth performance of the breeding animals is improved, the resistance of the breeding animals in a high-density breeding mode is enhanced, and the breeding method is high in operability. By adding the enhancing agent disclosed by the invention to the feed of aquatic animals, the survival rate of the aquatic animals is obviously improved compared with a test group without the enhancing agent.
The method of using the enhancer of the invention, and additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
Example 1
An enhancer for improving the immunity of aquatic animals comprises the following components in mass concentration: 15g/L of concentrated molasses fermentation liquor, 4g/L of melatonin, 15g/L of sodium alginate, 5g/L of nitrogen source, 3g/L of phosphorus source, 1g/L of potassium source, 3g/L of high-activity humic acid, 10 mu g/L of selenium methionine and 2g/L of composite EM bacteria.
The preparation method of the reinforcing agent comprises the following steps:
(1) preparing high-activity humic acid: adding humic acid into a potassium hydroxide solution (prepared from potassium hydroxide and water in a weight ratio of 1: 8) according to a weight ratio of 1:6 to dissolve, heating to 80 ℃, reacting for 40min under a pressure of 1Pa, cooling to room temperature, and continuing to hydrolyze for 5h to obtain the high-activity humic acid solution.
Preparation of potassium source: the potassium hydroxide and the potassium nitrate are mixed according to the mass ratio of 1: 2;
(2) according to the using amount of the components, the concentrated molasses fermentation liquor is added into the high-activity humic acid solution, and then the nitrogen source, the phosphorus source, the potassium source, the melatonin, the selenium methionine, the sodium alginate and the compound EM bacteria are added and mixed uniformly to obtain the reinforcing agent.
Example 2
An enhancer for improving the immunity of aquatic animals comprises the following components in mass concentration: 18g/L of concentrated molasses fermentation liquor, 6g/L of melatonin, 18g/L of sodium alginate, 7g/L of nitrogen source, 4g/L of phosphorus source, 1.5g/L of potassium source, 4g/L of high-activity humic acid, 12 mu g/L of selenium methionine and 6g/L of composite EM bacteria.
The preparation method of the reinforcing agent comprises the following steps:
(1) preparing high-activity humic acid: adding humic acid into a potassium hydroxide solution (prepared from potassium hydroxide and water in a weight ratio of 1: 9) according to a weight ratio of 1:8 to dissolve, heating to 90 ℃, reacting for 50min under a pressure of 1.5Pa, cooling to room temperature, and continuously hydrolyzing for 5.5h to obtain a high-activity humic acid solution.
Preparation of potassium source: potassium hydroxide and potassium nitrate are mixed according to the ratio of 1: 2.5;
(2) according to the using amount of the components, the concentrated molasses fermentation liquor is added into the high-activity humic acid solution, and then the nitrogen source, the phosphorus source, the potassium source, the melatonin, the selenium methionine, the sodium alginate and the compound EM bacteria are added and mixed uniformly to obtain the reinforcing agent.
Example 3
An enhancer for improving the immunity of aquatic animals comprises the following components in mass concentration: 20g/L of concentrated molasses fermentation liquor, 8g/L of melatonin, 20g/L of sodium alginate, 8g/L of nitrogen source, 5g/L of phosphorus source, 2g/L of potassium source, 5g/L of high-activity humic acid, 15 mu g/L of selenium methionine and 10g/L of composite EM bacteria.
The preparation method of the reinforcing agent comprises the following steps:
(1) preparing high-activity humic acid: adding humic acid into a potassium hydroxide solution (prepared from potassium hydroxide and water in a weight ratio of 1: 10) according to a weight ratio of 1:10 to dissolve, heating to 100 ℃, reacting for 60min under a pressure of 2Pa, cooling to room temperature, and continuously hydrolyzing for 6h to obtain a high-activity humic acid solution.
Preparation of potassium source: the potassium hydroxide and the potassium nitrate are mixed according to the mass ratio of 1: 3;
(2) according to the using amount of the components, the concentrated molasses fermentation liquor is added into the high-activity humic acid solution, and then the nitrogen source, the phosphorus source, the potassium source, the melatonin, the selenium methionine, the sodium alginate and the compound EM bacteria are added and mixed uniformly to obtain the reinforcing agent.
Example 4
The experimental feed prepared by adding the components and the preparation method of the enhancer for improving the immunity of the aquatic animals disclosed in the embodiment 3 into the feed for cultivating the aquatic animals is specifically described in the following by the application to the cultivation of crayfish.
1. Test materials
1.1 test reagents
L-DOPA (levodopa, which measures phenol oxidase activity), an acid phosphatase assay kit (WST-1 method), a total superoxide dismutase assay kit (WST-1 method), and a basal feed for crayfish (basal feed prepared under application No. CN 2016105910271).
1.2 test feed
Test feed A: the dose of immunopotentiator (example 1) added to the basal diet was 10mL/kg of diet;
test feed B: the dose of immunopotentiator (example 2) added to the basal diet was 20mL/kg of diet;
test feed C: the dose of immunopotentiator (example 3) added to basal diet was 15mL/kg of diet;
control feed: a basal feed.
1.3 Experimental Equipment
The device comprises an ultrafine grinder, an enzyme-labeling instrument and an electronic scale.
1.4 test crayfish
Healthy crayfish larvae with undamaged appearance, uniform size and 10 g/tail specification and taken from a crayfish breeding base.
2. Test method
2.1 test grouping:
dividing the test group into a test group A, a test group B, a test group C and a control group according to test requirements, wherein each group is provided with 3 repetitions, and 20 young crayfish are thrown in each repetition.
2.2 test field:
the test site adopts 12 aquatic culture tanks with the length, width and height of 120cm multiplied by 60cm, and the water depth is 40 cm. Before the test is started, plastic pipes are placed in the culture tanks to provide habitat for the crayfishes; after various indexes of the detected water quality are normal and stable, feeding the crayfish larvae in groups according to the above steps, and setting a feeding table to observe the feeding condition of the crayfish.
2.3 Breeding management
And (3) temporarily culturing for 7 days after seedling placement, feeding with basic feed, feeding with the experimental group A, feeding with the experimental group B, feeding with the experimental group C and feeding with the basic feed with the control group after the temporary culture period is finished. During the test period, the water temperature is controlled to be 32-35 ℃, the daily feeding amount of each group is 3% of the weight of the crayfish, the crayfish is fed for 2 times every day, the crayfish is fed at 7 o' clock in the morning, and the daily feeding amount is 30%; feeding at 7 pm, wherein the daily feeding amount is 70%, the feeding amount is adjusted in time according to the feeding and growth conditions of the crayfishes during the feeding period, and the water level and the water quality regulation and control are the same as those in the temporary culture process.
2.4 detection sample Collection
After the test is started, sampling is carried out on the 28 th day, and the sampling steps are as follows: randomly fishing 3 shrimps in each culture tank, measuring the weight of all shrimps, extracting hemolymph from a pericardial cavity by using a 1mL syringe, putting the shrimps in a refrigerator at 4 ℃ overnight, taking out the shrimps, centrifuging the shrimps for 10min by using a 4 ℃ centrifuge (5000r/min), taking supernatant (serum), and freezing and storing the supernatant at minus 20 ℃ for later use.
2.5 evaluation of Effect
3 indexes of acid phosphatase, superoxide dismutase and phenol oxidase were selected to evaluate immune function, wherein the acid phosphatase and superoxide dismutase activities were measured using corresponding kits, and the phenol oxidase activity was measured using a modified Ashida method.
3. And (3) test results:
3.1 immune function test:
the results of changes in acid phosphatase, superoxide dismutase, and phenol oxidase in test group a, test group B, test group C, and control group are shown in table 1, respectively, and it can be seen from table 1 that the activities of acid phosphatase, superoxide dismutase, and phenol oxidase in test group A, B and group C were significantly improved at day 28 as compared with the control group.
TABLE 1 Effect of immunopotentiators on the antioxidant capacity of juvenile crayfishes
As can be seen from Table 1, the enhancer for enhancing the immunity of crayfishes in high-temperature seasons can significantly enhance the immunity of crayfishes.
3.2 survival assessment:
after 28 days of experiment, the survival rates of crayfish in the control group, the experiment group a, the experiment group B and the experiment group C were respectively: 63.33%, 91.67%, 95.00% and 90.00%.
Therefore, the enhancer for improving the immunity of the aquatic animals can obviously enhance the immunity of crayfishes and can obviously improve the survival rate of the crayfishes in high-temperature seasons.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (9)
1. The enhancer for improving the immunity of aquatic animals is characterized by comprising the following components in mass concentration: 15-20g/L of concentrated molasses fermentation liquor, 4-8g/L of melatonin, 15-20g/L of sodium alginate, 5-8g/L of nitrogen source, 3-5g/L of phosphorus source, 1-2g/L of potassium source, 3-5g/L of high-activity humic acid and 10-15 mu g/L of selenium methionine.
2. The enhancer for improving immunity of aquatic animals according to claim 1, further comprising 2-10g/L of EM complex bacteria.
3. The enhancer for enhancing immunity of aquatic animals according to claim 1, wherein said nitrogen source is urea; the phosphorus source is monoammonium phosphate; the potassium source is a mixture of potassium hydroxide and potassium nitrate in a mass ratio of 1: 2-3.
4. The enhancer for improving the immunity of aquatic animals according to claim 1, wherein the high-activity humic acid is prepared by the following method: adding humic acid into an alkali solution according to the weight ratio of 1:6-1:10 for dissolving, heating to 80-100 ℃, reacting for 40-60min under the pressure of 1-2Pa, cooling to room temperature, and continuously hydrolyzing for 5-6h to obtain the high-activity humic acid.
5. The enhancer for improving the immunity of aquatic animals according to claim 4, wherein the alkali solution is a potassium hydroxide solution, and the potassium hydroxide solution is prepared by mixing potassium hydroxide and water in a weight ratio of 1:8-1: 10.
6. A method for preparing an enhancer for enhancing immunity of aquatic animals according to any one of claims 1-5, comprising the steps of: adding the concentrated molasses fermentation liquor into the high-activity humic acid solution according to the component dosage of the enhancer for improving the immunity of the aquatic animals, adding the nitrogen source, the phosphorus source, the potassium source, the melatonin, the selenium methionine, the sodium alginate and the compound EM bacteria, and uniformly mixing to obtain the enhancer.
7. Use of an enhancer for enhancing the immunity of an aquatic animal according to any one of claims 1 to 5 in a basal feed for the cultivation of an aquatic animal.
8. The use according to claim 7, wherein the enhancing agent is added to the basal feed at a volume concentration of 10-20 mL/kg.
9. The use of claim 7, wherein the enhancing agent is sprayed uniformly onto the surface of the basal feed by spraying and then dried in the shade for 45-60min to obtain the experimental feed.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113180143A (en) * | 2021-05-24 | 2021-07-30 | 北京博思汇科生物技术有限公司 | Method for producing organic trace humic acid selenium of bacterial liquid additive |
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