阅读说明：本技术 一种提高鱼抗应激反应天然组合物及其应用 (Natural composition for improving fish anti-stress response and application thereof ) 是由 李华涛 袁佳 杜文浩 吴敏 敬晓琴 罗朋 龙娇 刘思妙 冯敏 袁志 李绮绮 周 于 2020-07-23 设计创作，主要内容包括：本发明公开了一种提高鱼抗应激反应天然组合物,包括天然复合抗应激剂I和/或天然复合抗应激剂II；所述天然复合抗应激剂I包括丹参有机溶剂提取物、紫苏叶有机溶剂提取物和当归有机溶剂提取物中的两种或两种以上；所述天然复合抗应激剂II包括丹参水提取物、银杏叶水提取物和桑叶水提取物中的两种或两种以上。本发明提供的天然组合物用于制备鱼抗缺氧再复氧应激反应添加剂或饲料,为天然植物提取物,基础饲料原料没有人为添加其他物质。此天然功能性饲料能提高养殖中鱼对缺氧的耐受力和复氧成活率,无残留、无毒副作用、不污染环境。(The invention discloses a natural composition for improving fish anti-stress response, which comprises a natural compound anti-stress agent I and/or a natural compound anti-stress agent II; the natural compound anti-stress agent I comprises two or more than two of salvia miltiorrhiza organic solvent extract, perilla leaf organic solvent extract and angelica sinensis organic solvent extract; the natural compound anti-stress agent II comprises two or more of water extract of Saviae Miltiorrhizae radix, water extract of folium Ginkgo and water extract of folium Mori. The natural composition provided by the invention is used for preparing the anti-hypoxia reoxygenation stress response additive or feed for the fish, is a natural plant extract, and other substances are not artificially added into the basic feed raw materials. The natural functional feed can improve the tolerance of the cultured fish to oxygen deficiency and the reoxygenation survival rate, and has no residue, no toxic or side effect and no environmental pollution.)

1. A natural composition for improving fish anti-stress response is characterized by comprising a natural compound anti-stress agent I and/or a natural compound anti-stress agent II; the natural compound anti-stress agent I comprises two or more than two of salvia miltiorrhiza organic solvent extract, perilla leaf organic solvent extract and angelica sinensis organic solvent extract; the natural compound anti-stress agent II comprises two or more of water extract of Saviae Miltiorrhizae radix, water extract of folium Ginkgo and water extract of folium Mori.

2. The natural composition for improving fish anti-stress response as claimed in claim 1, wherein the organic solvent extract of red sage root is an ethyl acetate extract of red sage root, the organic solvent extract of perilla leaf is an ethyl acetate extract of perilla leaf, and the organic solvent extract of angelica is an ethyl acetate extract of angelica.

3. The natural composition for improving the anti-stress response of fish as claimed in claim 1, wherein the natural compound anti-stress agent I comprises three kinds of organic solvent extracts of radix salviae miltiorrhizae, folium perillae and radix angelicae sinensis, and the mass ratio of the organic solvent extracts of radix salviae miltiorrhizae, folium perillae and radix angelicae sinensis is 2:1:1 or 1:2:1 or 1:1:2 or 2:2:1 or 1:2:2 or 2:1: 2.

4. The natural composition for improving the anti-stress response of fish as claimed in claim 1, wherein the natural compound anti-stress agent II comprises three kinds of water extracts of radix salviae miltiorrhizae, folium ginkgo and folium mori, and the mass ratio of the water extracts of radix salviae miltiorrhizae, folium ginkgo and folium mori is 2:1:1 or 1:2:1 or 1:1:2 or 2:2:1 or 1:2:2 or 2:1: 2.

5. The natural composition for improving fish anti-stress response as claimed in claim 1, wherein the mass ratio of the natural compound anti-stress agent I to the natural compound anti-stress agent II is 0.5-2.0.

6. Use of a natural composition for enhancing the anti-stress response of fish as claimed in any one of claims 1 to 5, for the preparation of an anti-stress response additive or feed for freshwater fish; the stress response includes an anoxic reoxygenation stress response.

7. A fish feed comprising a basal feed and a natural composition as claimed in any one of claims 1 to 7 for enhancing the anti-stress response of fish.

8. The fish feed as claimed in claim 7, wherein the natural compound anti-stress agent I is added in an amount of 0.3-0.5% by weight based on the weight of the basic feed; the addition amount of the natural compound anti-stress agent II is 0.5 to 1.0 percent of the weight of the basic feed.

9. The fish feed of claim 8, wherein the basic feed comprises the following components in parts by weight: the basic feed comprises the following components in parts by weight: 15-19 parts of fish meal, 20-25 parts of soybean meal, 15-17 parts of peanut meal, 3-7 parts of cottonseed meal, 0.3-0.7 part of DL-methionine, 32-36 parts of wheat flour, 1.0-1.5 parts of rapeseed oil, 0.5-1.0 part of vitamin compound and 0.5-1.0 part of mineral compound.

10. A process for preparing a fish feed as claimed in any one of claims 7 to 9, wherein the natural complex anti-stress agent I is dissolved in fat and mixed, and then added to the basal feed and mixed; and/or dissolving natural compound anti-stress agent II in water, mixing, adding into basic feed, and mixing.

Technical Field

The invention relates to the technical field of freshwater fish culture, in particular to a natural composition for improving anti-stress reaction of fish and application thereof.

Background

The current situation of aquaculture is mainly natural water body culture, and freshwater culture is mainly pond culture. The growth of farmed fish depends on the dissolved oxygen content in the water body. However, natural water body cultivation has the disadvantage that dissolved oxygen in water body is greatly influenced by environmental factors. The dissolved oxygen content in natural water is affected by day and night, weather and seasonal changes. The photosynthesis of green plants in the water body is very active in sunny days, and the dissolved oxygen content of the water body may reach an oversaturated state; the dissolved oxygen in the water body is greatly reduced due to the stagnation of the photosynthesis of the plants at night. Weather changes can affect illumination and photosynthesis of plants, thereby affecting dissolved oxygen in water. Seasonal changes can affect not only the light but also the water temperature. In the physiological temperature range of the fish, the oxygen consumption of the fish body is increased by 2-3 times when the water temperature is increased by 10 ℃. It has been reported that the fluctuation of dissolved oxygen in natural water body can make fish body suffer from oxygen deficiency and reoxygenation stress, so that the production performance of fish is reduced.

"high density, intensification" is an important feature of current aquaculture. But the fish carrying amount of the water body is improved by the high-density culture mode; the relative living space of the fishes in the water body is reduced, the competition and interference among different individuals are intensified, the activity of the fishes is improved, and the factors improve the oxygen demand of the fishes to different degrees. The high-density culture improves the feed feeding amount, increases culture wastes in the water body, and aggravates the conversion of water body substances, and the factors improve the oxygen consumption in the water body. The above situation further aggravates the anaerobic stress of the cultured fish. The oxygen-increasing machine can greatly relieve the oxygen deficiency state of the high-density aquaculture water body, but also aggravates the possibility that the fish body suffers reoxygenation stress. The phenomenon of pond turning over and fish death caused by the change of dissolved oxygen in the water body still often occurs. The anoxia and reoxygenation stress reaction can reduce the immunity of the fish and improve the morbidity and mortality of the fish, thereby improving the use amount of antibiotics and chemical drugs in the culture and increasing the drug residue of fish meat. The defects of natural water body culture and high-density culture make the market urgently need a feed which can improve the anti-hypoxia and reoxygenation stress capability of the fish without residue, toxic or side effect and environmental pollution, and is used for improving the culture benefit, reducing the drug residue and improving the fish meat quality.

High-density intensive culture requires the use of complete formula feed. The addition of antioxidants, mildewcides, growth promoters, phagostimulants and the like to compound feeds is a common phenomenon. The major antioxidants currently used are mainly synthetic antioxidants, including Ethoxyquinoline (EQ), Butylhydroxyanisole (BHA), dibutylhydroxytoluene (BHT), and the like. The fish meal and the grease are important feed raw materials, and the adding amount of the synthetic antioxidant in the fish meal processing process can reach 0.1 percent; synthetic antioxidants are also added to feed-grade oils. However, synthetic antioxidants have been reported to be carcinogenic and genotoxic to animals and to cause residues and enrichment in fish products, creating a food safety hazard and endangering human health. In 2 months in 2017, the European Union issues the G/SPS/EU/190 announcement in 2017: the addition of EQ to the animal feed was stopped. Also, BHT and BHA added to animal feed have been reported to have higher carcinogenic risk than EQ. Chemically synthesized additives such as mildewcides, growth promoters, phagostimulants and the like have also been reported to be potentially toxic to animals and may cause residues in fish meat. The disadvantages of compound feeds will therefore lead to an increased market demand for natural product compound feeds.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: the invention provides a natural composition for improving fish anti-stress response and an application thereof, wherein the natural composition can solve the problems that fish are subjected to hypoxia reoxygenation stress aggravated by high-density and natural water body culture, the existing feed additives have toxic and side effects and even carcinogenic effects on animals, and can cause food safety hidden danger and human health danger caused by residue in fish meat. The compound is a natural compound prepared based on medicinal and edible Chinese herbal medicines, is used as an anti-hypoxia reoxygenation stress response additive for fish and for preparing natural functional feed, has no toxic or side effect, and has important significance for improving the economic and social benefits of the fish.

The invention is realized by the following technical scheme:

a natural composition for improving fish anti-stress response comprises natural compound anti-stress agent I and/or natural compound anti-stress agent II; the natural compound anti-stress agent I comprises two or more than two of salvia miltiorrhiza organic solvent extract, perilla leaf organic solvent extract and angelica sinensis organic solvent extract; the natural compound anti-stress agent II comprises two or more of water extract of Saviae Miltiorrhizae radix, water extract of folium Ginkgo and water extract of folium Mori.

The fish anti-stress reaction compound provided by the invention is a natural plant extract, and other substances are not artificially added. The natural functional feed prepared by taking the active substance can improve the tolerance of the cultured fish to oxygen deficiency and the reoxygenation survival rate, and has no residue, no toxic or side effect and no environmental pollution.

Preferably, the salvia miltiorrhiza organic solvent extract is salvia miltiorrhiza ethyl acetate extract, the perilla leaf organic solvent extract is perilla leaf ethyl acetate extract, and the angelica sinensis organic solvent extract is angelica sinensis ethyl acetate extract.

Preferably, the content of flavonoids in the salvia miltiorrhiza ethyl acetate extract is 82.53mg/g-97.60 mg/g; the content of flavonoids in the perilla leaf ethyl acetate extract is 64.45mg/g-70.28 mg/g; the content of polyphenols in the radix Angelicae sinensis ethyl acetate extract is 46.76mg/g-55.31 mg/g.

The red sage root ethyl acetate extract is obtained by extracting analytical grade ethyl acetate and is a brownish red extract, wherein the content of flavonoids is 82.53mg/g-97.60 mg/g. The folium Perillae ethyl acetate extract is obtained by extracting with analytical grade ethyl acetate, and is dark brown extract, wherein the content of flavonoids is 64.45mg/g-70.28 mg/g. The angelica ethyl acetate extract is obtained by extracting analytical grade ethyl acetate, and the extract is a brownish red extract, wherein the content of polyphenol substances is 46.76mg/g-55.31 mg/g.

Further preferably, in the salvia miltiorrhiza water extract, the mass percentage of crude protein is 14.23% -16.08%, and the mass percentage of soluble sugar is 38.89% -46.30%; in the ginkgo leaf water extract, the mass percentage of crude protein is 5.76-7.14%, and the mass percentage of soluble sugar is 51.32-58.29%; in the mulberry leaf water extract, the mass percentage of crude protein is 13.07-15.64%, and the mass percentage of total sugar is 35.67-42.54%.

The red sage root water extract is obtained by extracting with distilled water and is brownish red powder, the mass percentage of crude protein is 14.23-16.08 percent, and the mass percentage of soluble sugar is 38.89-46.30 percent. The water extract of mulberry leaves is obtained by extracting with distilled water and is dark brown powder, the mass percentage of crude protein is 13.07-15.64 percent, and the mass percentage of total sugar is 35.67-42.54 percent. The ginkgo leaf water extract is obtained by extracting through distilled water and is light brown powder, the mass percentage of crude protein is 5.76-7.14%, and the mass percentage of soluble sugar is 51.32-58.29%.

Preferably, the natural compound anti-stress agent I comprises three kinds of salvia miltiorrhiza organic solvent extract, perilla leaf organic solvent extract and angelica sinensis organic solvent extract, and the mass ratio of the salvia miltiorrhiza organic solvent extract, the perilla leaf organic solvent extract and the angelica sinensis organic solvent extract is 2:1:1 or 1:2:1 or 1:1:2 or 2:2:1 or 1:2:2 or 2:1: 2.

Further preferably, the natural compound anti-stress agent II comprises three of a salvia miltiorrhiza water extract, a ginkgo biloba leaf water extract and a mulberry leaf water extract, and the mass ratio of the salvia miltiorrhiza water extract, the ginkgo biloba leaf water extract and the mulberry leaf water extract is 2:1:1 or 1:2:1 or 1:1:2 or 2:2:1 or 1:2:2 or 2:1: 2.

Further preferably, the mass ratio of the natural compound anti-stress agent I to the natural compound anti-stress agent II is 0.3-5.

The application of the natural composition for improving the anti-stress response of the fish is used for preparing an anti-stress response additive or feed for freshwater fish; the stress response includes an anoxic reoxygenation stress response. The feed herein is preferably a natural functional feed.

A fish feed comprises a basal feed and the natural composition for improving the anti-stress response of fish.

Further preferably, the adding amount of the natural compound anti-stress agent I is 0.3 to 0.5 percent of the weight of the basic feed; the addition amount of the natural compound anti-stress agent II is 0.5 to 1.0 percent of the weight of the basic feed.

Further preferably, the basic feed comprises the following components in parts by weight: 15-19 parts of fish meal, 20-25 parts of soybean meal, 15-17 parts of peanut meal, 3-7 parts of cottonseed meal, 0.3-0.7 part of DL-methionine, 32-36 parts of wheat flour, 1.0-1.5 parts of rapeseed oil, 0.5-1.0 part of vitamin compound and 0.5-1.0 part of mineral compound. Preferably, the basic feed comprises the following components in parts by weight: 17 parts of fish meal, 23 parts of soybean meal, 16 parts of peanut meal, 5 parts of cottonseed meal, 0.5 part of DL-methionine, 35 parts of wheat flour, 1.5 parts of rapeseed oil, 1 part of vitamin compound and 1 part of mineral compound.

The main raw materials of the anti-fish hypoxia reoxygenation feed are all natural products, and other substances are not artificially added; the additive components meet the requirements of national standard NY/T471-2018; the functional components of the feed additive are natural plant extracts, and the feed additive has certain functions of resisting feed oxidation and preventing mildew. The natural functional feed can improve the tolerance of the fish to oxygen deficiency and the reoxygenation survival rate in the culture.

The preparation method of the fish feed comprises the steps of dissolving the natural compound anti-stress agent I in grease, mixing, adding into the basic feed, and mixing; and/or dissolving natural compound anti-stress agent II in water, mixing, adding into basic feed, and mixing.

For example, in preparing a feed containing natural compound anti-stress agent I and natural compound anti-stress agent II: firstly, calculating the mass of oil required by unit mass of basic feed according to a feed formula, adding a natural compound anti-stress agent I accounting for 0.3-0.5% of the weight of the basic feed into the oil, and then uniformly mixing the oil and the basic feed; and dissolving the natural compound anti-stress agent II with the weight of 0.5-1.0% of the weight of the basic feed into drinkable water, and uniformly mixing the dissolved natural compound anti-stress agent II into the basic feed to prepare pellet feed for inhibiting the anoxic reoxygenation stress reaction of the freshwater fish.

The invention has the following advantages and beneficial effects:

1. the fish anti-stress reaction compound provided by the invention is a natural plant extract, and other substances are not artificially added. The natural functional feed prepared by taking the active substance can improve the tolerance of the cultured fish to oxygen deficiency and the reoxygenation survival rate, and has no residue, no toxic or side effect and no environmental pollution.

2. The invention adopts the extract of natural plants with homology of medicine and food as the feed additive, removes most ineffective components to enhance the activity, and the comprehensive functions of the medicinal plants are decomposed after extraction, and the specific functions are concentrated, thereby improving the effect of a certain function. The plant extract has reduced volume compared with Chinese herbal medicine, and is convenient for storage, transportation and transaction.

Drawings

The accompanying drawings, which are included to provide a further understanding of the embodiments of the invention and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the invention and together with the description serve to explain the principles of the invention. In the drawings:

FIG. 1 is a graph showing the effect of feeding compound ester extract on the duration of hypoxia tolerance of crucian; in the figure, data are presented as mean ± standard deviation of 3 replicates; the difference in the upper case letters in the same column data indicates significant difference (P < 0.05);

fig. 2 shows the effect of feeding compound ester extract on the reoxygenation recovery of crucian after oxygen deficiency and side turning; in the figure, data are presented as mean ± standard deviation of 3 replicates; the difference in the upper case letters in the same column data indicates significant difference (P < 0.05);

FIG. 3 is the effect of the feed of the compound water extract on the duration of hypoxia tolerance of crucian; in the figure, data are presented as mean ± standard deviation of 3 replicates; the difference in the upper case letters in the same column data indicates significant difference (P < 0.05);

FIG. 4 shows the effect of feeding the feed with the compound water extract on the reoxygenation recovery of crucian after oxygen deficiency and rollover; in the figure, data are presented as mean ± standard deviation of 3 replicates; the difference in the upper case letters in the same column data indicates significant difference (P < 0.05);

FIG. 5 is the effect of feeding compound anti-stress agent feed on the duration of hypoxia tolerance of crucian; in the figure, data are presented as mean ± standard deviation of 3 replicates; the difference in the upper case letters in the same column data indicates significant difference (P < 0.05);

fig. 6 shows the effect of feeding a compound anti-stress agent on the reoxygenation recovery of crucian after oxygen deficiency and side turning; in the figure, data are presented as mean ± standard deviation of 3 replicates; the difference in the upper-case letters in the same column data indicates significant difference (P < 0.05).

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples and accompanying drawings, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not meant to limit the present invention.