CN111772057A - Natural compound for improving fish anti-stress response and application thereof - Google Patents
Natural compound for improving fish anti-stress response and application thereof Download PDFInfo
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Abstract
The invention discloses a natural compound 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 of wild licorice organic solvent extract, angelica organic solvent extract and mulberry leaf organic solvent extract; the natural compound anti-stress agent II comprises two or more of water extract of fructus crataegi, water extract of radix Angelicae sinensis and water extract of herba Leonuri. The application of the natural compound 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 a high density stress response. The fish anti-stress reaction compound provided by the invention is a natural plant extract, and other substances are not artificially added to the basic feed raw materials. The natural functional feed can inhibit fish weight gain, feed intake and feed efficiency reduction caused by high density stress of culture, and has no residue, no toxic and side effects, and no environmental pollution.
Description
Technical Field
The invention relates to the technical field of freshwater fish culture, in particular to a natural compound for improving anti-stress reaction of fish and application thereof.
Background
"high density, intensification" is an important feature of current aquaculture. However, the living space of the fishes in the water body is reduced in the high-density culture mode, and competition and interference among different individuals are aggravated; the feed feeding amount is improved, the culture wastes in the water body are increased, and the water quality is deteriorated; these factors increase the stress response of the fish, thereby reducing the growth rate, feed intake and feed efficiency of the fish. Stress reactions can also reduce fish immunity and increase fish morbidity and mortality. The factors also improve the use amount of antibiotics and chemical drugs in the culture, so that the drug residue of the fish meat is increased. The defects of the high-density culture mode make the market urgently need a feed which can improve the anti-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 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; 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 compound for improving the stress response of fish and an application thereof, wherein the natural compound is prepared based on medicinal and edible Chinese herbal medicines, is used as a natural stress response resisting additive for fish and feed, has no toxic or side effect, and has important significance for improving the economic and social benefits of the natural compound.
The invention is realized by the following technical scheme:
a natural compound 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 of wild licorice organic solvent extract, angelica organic solvent extract and mulberry leaf organic solvent extract; the natural compound anti-stress agent II comprises two or more of water extract of fructus crataegi, water extract of radix Angelicae sinensis and water extract of herba Leonuri.
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 adding the basic feed can inhibit fish weight gain, food intake and feed efficiency reduction caused by high-density stress of culture, and has no residue, no toxic or side effect and no environment pollution.
Preferably, the wild licorice organic solvent extract is a wild licorice ethanol extract, the angelica organic solvent extract is an angelica ethyl acetate extract, and the mulberry leaf organic solvent extract is a mulberry leaf ethyl acetate extract.
Further preferably, the content of polyphenols in the angelica ethyl acetate extract is 47.21mg/g-54.60 mg/g; the content of polyphenol substances in the mulberry leaf ethyl acetate extract is 70.87mg/g-74.55 mg/g; the content of flavonoids in the wild licorice ethanol extract is 61.89mg/g-68.72 mg/g.
The ethyl acetate extract of radix Angelicae sinensis is brownish red extract, wherein the content of polyphenols is 47.21mg/g-54.60 mg/g; the mulberry leaf ethyl acetate extract is dark green extract, wherein the content of polyphenols is 70.87mg/g-74.55 mg/g; the wild licorice ethanol extract is a dark green extract, wherein the content of flavonoids is 61.89mg/g-68.72 mg/g.
Further preferably, in the Chinese angelica water extract, the mass percentage of crude protein is 19.05-22.56%, and the mass percentage of soluble sugar is 31.64-35.15%; in the hawthorn water extract, the mass percentage of crude protein is 15.19-18.33%, and the mass percentage of soluble sugar is 37.35-41.90%; in the motherwort water extract, the mass percentage of crude protein is 12.16-15.29%, and the mass percentage of soluble sugar is 28.30-32.42%.
The water extract of the angelica is light red yellow powder, wherein, the mass percentage of crude protein is 19.05 percent to 22.56 percent, and the mass percentage of soluble sugar is 31.64 percent to 35.15 percent; the hawthorn water extract is light red brown powder, wherein the mass percentage of crude protein is 15.19-18.33%, and the mass percentage of soluble sugar is 37.35-41.90%; the motherwort water extract is a grey brown powder, wherein the mass percentage of crude protein is 12.16-15.29%, and the mass percentage of soluble sugar is 28.30-32.42%.
Further preferably, the natural composite anti-stress agent I comprises three kinds of wild licorice organic solvent extract, Chinese angelica organic solvent extract and mulberry leaf organic solvent extract, and the mass ratio of the wild licorice organic solvent extract to the Chinese angelica organic solvent extract to the mulberry leaf organic solvent extract is (1-2): (1-2), preferably 1:1: 1.
Further preferably, the natural compound anti-stress agent II comprises three of a hawthorn water extract, an angelica water extract and a leonurus water extract, and the mass ratio of the hawthorn water extract to the angelica water extract to the leonurus water extract in the natural compound anti-stress agent II is (1-2): 1-2, preferably 1:1: 1.
Further preferably, the mass ratio of the natural compound anti-stress agent I to the natural compound anti-stress agent II is 0.5-2.
The application of the natural compound 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 a high density stress response. Preferably, the feed is a natural functional feed.
A fish feed comprises a basal feed and the natural compound 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 natural compound anti-stress agent I is dissolved in grease and mixed, and then is added into the basic feed raw material and mixed; and/or dissolving natural compound anti-stress agent II in water, mixing, adding into basic feed raw material, 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 accounting for 0.5-1.0 percent 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 raw materials to prepare granulated feed for inhibiting the high-density stress reaction of the freshwater fish.
Further preferably, the basic feed raw material is a natural feed. The basic feed comprises the following components in parts by weight: 15-20 parts of fish meal, 20-25 parts of soybean meal, 14-18 parts of peanut meal, 4-6 parts of cottonseed meal, 0.5-1.0 part of DL-methionine, 30-35 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.
Wherein, the fish meal, the bean pulp, the peanut pulp, the cottonseed meal, the wheat flour and the rapeseed oil are not artificially added with other substances except the substances contained in the fish meal, the bean pulp, the peanut pulp, the cottonseed meal, the wheat flour and the rapeseed oil; the components of the DL-methionine, the vitamin additive and the mineral additive meet the requirements of national standard (NY/T471-2018) green food feed and feed additive use criteria.
The invention has the following advantages and beneficial effects:
1. the invention provides a natural compound for improving the fish anti-stress response and an application thereof, which solve the problems, the natural compound is prepared based on medicinal and edible Chinese herbal medicines, is used as an anti-stress response additive for fish, is a natural plant extract, and is free from other substances artificially added. The natural functional feed can inhibit fish weight gain, food intake and feed efficiency reduction caused by high-density stress of culture, and has no residue, toxic or side effect and no environmental pollution; has important significance for improving the economic and social benefits.
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 of the effect of feed density on fish weight gain; data are presented as mean ± standard deviation of 3 replicates; the letters marked on the bars indicate that the difference between the data is significant, and the same letters indicate that the difference is not significant; weight gain was final weight-initial weight (table 1).
FIG. 2 is a graph of the effect of feed density on fish feed efficiency; data are presented as mean ± standard deviation of 3 replicates; the letters marked on the bars indicate significant differences between the data, while the same indicate insignificant differences. Feed efficiency is 100 x gain/feed intake;
fig. 3 shows weight gain recovery rate of crucian carp after feeding compound alcohol ester extract feed for 28 days; data are presented as mean ± standard deviation of 3 replicates. Weight gain recovery (%) - (a-Y)/(K-Y) × 100 (table 4);
FIG. 4 shows the weight gain recovery rate of crucian carp after feeding compound water extract feed for 28 days; data are presented as mean ± standard deviation of 3 replicates. Weight gain recovery (%) - (a-Y)/(K-Y) × 100 (table 6);
FIG. 5 shows the weight gain recovery rate of crucian carp after feeding compound anti-stress agent feed for 28 days; data are presented as mean ± standard deviation of 3 replicates. Weight gain recovery (%) - (a-Y)/(K-Y) × 100 (table 8).
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.
Examples
The embodiment provides a natural compound for improving the anti-stress response of fish and application thereof.
Compound for improving fish anti-stress reaction
1) Organic solvent alcohol ester extract
The angelica ethyl acetate extract is obtained by analytical grade ethyl acetate extraction, is a brownish red extract, and has the polyphenol substance content of 47.21mg/g-54.60 mg/g.
The mulberry leaf ethyl acetate extract is obtained by extracting analytical grade ethyl acetate, is dark green extract, and has the polyphenol substance content of 70.87mg/g-74.55 mg/g.
The wild licorice ethanol extract is obtained by analytical grade ethanol extraction, is dark green extract, and has the flavonoid content of 61.89mg/g-68.72 mg/g.
The hawthorn ethyl acetate extract is obtained by extracting analytical grade ethyl acetate, is a yellow brown extract, and has the flavonoid content of 103.26mg/g-117.3 mg/g.
The leonurus ethanol extract is obtained by extracting through analytical grade ethanol, is dark green extract, and has the flavonoid content of 66.75mg/g-74.37 mg/g.
2) Aqueous extracts
The water extract of angelica is obtained by extracting with distilled water, and is light red yellow powder, the crude protein content is 19.05-22.56%, and the soluble sugar content is 31.64-35.15%.
The water extract of mulberry leaves is obtained by extracting with distilled water and is dark brown powder, the content of crude protein is 13.07-15.64%, and the content of total sugar is 33.26-37.77%.
The wild licorice water extract is obtained by extracting with distilled water, and is dark brown powder, the crude protein content is 17.56% -20.03%, and the total sugar content is 28.32% -32.20%.
The hawthorn water extract is obtained by extracting through distilled water, and is light red brown powder, the content of crude protein is 15.19-18.33%, and the content of soluble sugar is 37.35-41.90%.
The water extract of motherwort is gray brown powder with crude protein content of 12.16-15.29% and soluble sugar content of 28.30-32.42%.
Second, basic feed
The used raw material sources are as follows: the fish meal, the bean pulp, the peanut meal, the cottonseed meal, the wheat flour and the rapeseed oil are all self-made, and other substances are not added artificially except the nutrients of the materials. The specific formulation is shown in table 1:
TABLE 1 basic feed formulation
Remarking:
1the fish meal contains 89.88% of dry matter, 51.70% of crude protein, 15.01% of crude fat and 18.63% of crude ash, and is prepared from low-cost silver carp, crucian carp and grass carp by air-blast drying at 80 deg.C.
2The soybean meal contains 89.13% of dry matter, 46.92% of crude protein, 1.53% of crude fat and 5.46% of crude ash, and is a byproduct of soybean after heating, crushing, oil extraction and solvent evaporation.
3The peanut meal contains 88.21% of dry matter, 47.54% of crude protein, 1.42% of crude fat and 5.33% of crude ash, and is a byproduct of peanut kernels after heating, crushing, oil extraction and solvent evaporation.
4The cottonseed meal contains 89.46% of dry matter, 44.37% of crude protein, 0.54% of crude fat and 5.82% of crude ash, and is a byproduct of cottonseed kernels after heating, crushing, oil extraction and solvent evaporation.
5DL-methionine meets the requirement of NY/T471-2018.
6Wheat flour is obtained by pulverizing wheat, and contains dry matter 87.60%, crude protein 13.32%, crude fat 2.03%, and crude ash 1.84%.
7Rapeseed oil contains dry matter 99.03%0.00% of crude protein, 98.00% of crude fat and 0.53% of crude ash, and is prepared by pressing rapeseed.
8Each kilogram of vitamin additive comprises 500,000IU of vitamin A acetate, D3240,000IU of vitamin, 20.00g of DL- α -tocopherol, 30.10g of vitamin K, 10.10 g of vitamin B, 0.50g of riboflavin, 0.92g of pyridoxine hydrochloride, 0.001g of cyanocobalamin, 2.46g of D-calcium pantothenate, 2.80g of nicotinic acid, 0.10g of D-biotin, 51.81g of inositol and 0.50g of folic acid, and the carrier is defatted rice bran, and each component meets the requirement of NY/T471-2018.
9The mineral supplement comprises per kilogram: FeSO437.90g, 1.69g of copper methionine, 23.71g of zinc methionine, 7.15g of manganese methionine, 0.06g of selenium methionine and 0.134g of KI, and the diluent is limestone powder. The components meet the requirements of NY/T471-2018.
Thirdly, screening of the optimal stress density
After crucian fries are purchased from the Yong Zhen in the city of Nejiang province in Sichuan province, the crucian fries are temporarily cultured in a culture room for one week. Selecting 540 tails of crucian carp fries with the weight of 14.1 +/-0.3 g, randomly dividing the crucian carp fries into 8 treatment groups, and 3 fish tanks in each group. The fish tails of 8 treatment groups were: 5. 10, 15, 20, 25, 30, 35 and 40 tails/cylinder. The shape, size and color of each fish tank are the same, the size is 30 multiplied by 40cm, and an 800L/h flow aerator is arranged in each fish tank. Drinkable tap water is used as a water source, and the volume of water in the jar is 30L. Controlling the culture room temperature to be 22 +/-1 ℃; the oxygen increasing machine continuously supplies oxygen; feeding fish meal and soybean meal type omnivorous fish feed; the water was changed every 2 days. The culture period is 28 days.
After the 28-day culture period, the weight gain, feed intake, feed efficiency and mortality of each treated group were examined and the results are shown in table 2 and figures 1-2. As can be seen from the figure, the stocking density with the highest weight increment is 0.17-0.50 tail/L water; the weight gain is gradually reduced along with the increase of the culture density, and the culture density with the minimum weight gain is 1.00-1.33 tails per L of water. The highest stocking density is 0.33-0.67 tail/L water; with the increase of the culture density, the food intake is gradually reduced, and the minimum density is 1.00-1.33 tail/L water. The stocking density with the highest feed efficiency is 0.17-0.50 tail/L water; with the increase of the breeding density, the feed efficiency is gradually reduced, and the density with the lowest feed efficiency is 1.00-1.33 tail/L water. When the stocking density is 1.17-1.33 fish/L water, the fish die. Namely, when the breeding density is 0.50 fish/L water, the weight gain, the food intake and the feed efficiency of the fish are highest, and the optimal breeding density is determined to be 0.50 fish/L water. As the density increases, the weight gain, feed intake and feed efficiency of the fish gradually decrease; when the density of the fish is increased to 1.00 tail/L water, the weight gain, food intake and feed efficiency of the fish are reduced to the lowest level; further improve the fish density to 1.17 fish/L water, and the fish die. The breeding density that produced the greatest density stress without causing fish death was 1.00 tails/L water.
Table 2 weight gain, feed intake, feed efficiency and mortality for each treatment group
Remarking: data in the table are presented as mean ± standard deviation of 3 replicates, 30L water per aquarium; in the same column of data, different superscript letters indicate that the data are different significantly, and the same letters indicate that the data are not different significantly.
Screening of anti-stress Compound combinations
1. Natural composite anti-stress agent I
According to the formula of the basic feed, 5 kinds of organic solvent extracts are respectively added into the grease of the basic feed at the concentration of 1g/kg according to the following table to prepare 6 kinds of corresponding pellet feeds. The experimental design was as follows:
TABLE 3 design of organic solvent extract anti-fish stress test
| Additive material | Blank space | Induction of | |
1 | 2 | 3 | 4 | 5 |
| Ethyl acetate extract of Chinese angelica | × | × | × | |||||
| Folium Mori ethyl acetate extract | × | × | × | |||||
| Fructus crataegi ethyl acetate extract | × | × | × | |||||
| Wild licorice ethanol extract | × | × | × | |||||
| Ethanol extract of motherwort | × | × | × | |||||
| Stocking density | 0.5 tail/ |
1 tail/ |
1 tail/ |
1 tail/ |
1 tail/ |
1 tail/ |
1 tail/ |
1 tail/L |
Remarking: the basal feed without any extract was labeled as "blank", the feed with all 5 extracts added was labeled as "control", and the feeds with the angelica ethyl acetate extract, the mulberry leaf ethyl acetate extract, the hawthorn ethyl acetate extract, the glycyrrhiza uralensis ethanol extract and the leonurus japonicus ethanol extract were subtracted one by one from the "control" and labeled as "1", "2", "3", "4" and "5", respectively.
Selecting 675 tails of crucian fries with the weight of 14.3 +/-0.3 g under the optimal stress density (1.00 tail/L water) and the same culture environment, randomly dividing the crucian fries into 8 treatment groups, wherein each group comprises 3 fish tanks, and each fish tank contains 30L of water. The fish tails of 8 treatment groups were: 15. 30, 30 and 30 tails/cylinder, designated "blank (K)", "induced (Y)", "control (a)" respectively0)”、“1(A1)”、“2(A2)”、“3(A3)”、“4(A4) "and" 5 (A)5) Group, fed with feed labeled "blank", "control", "1", "2", "3", "4" and "5", respectively. And measuring the growth, the food intake and the feed efficiency of the crucian after 28 days.
As shown in table 4 and fig. 3, the results showed that the growth, food intake and feed efficiency of the induced herds were significantly reduced compared to the blank group; however, feeding the complex extract restored fish growth, food intake and feed efficiency to a different extent compared to the induction group. The weight gain recovery rate is in the order of magnitude: 5 > 3 > control > 4 > 1 > 2, 79.79%, 71.43%, 57.72%, 45.01%, 35.36%, 23.70%, respectively. Therefore, in the components of the compound extract, the leonurus ethanol extract and the hawthorn ethyl acetate extract have an inhibiting effect on the anti-stress effect of other components of the compound extract; the wild licorice ethanol extract, the angelica ethyl acetate extract and the mulberry leaf ethyl acetate extract play a promoting role. Therefore, the components of the primarily screened compound extract are wild licorice ethanol extract, angelica ethyl acetate extract and mulberry leaf ethyl acetate extract, and are marked as compound anti-stress agent I.
TABLE 4 influence of feeding complex alcohol ester extract on crucian carp breeding density stress
Remarking: 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). Weight gain is equal to end weight-initial weight; feed efficiency was 100 x gain/feed intake.
2. Natural composite anti-stress agent II
According to the basal feed formulation, 5 extracts were added to the basal feed in a concentration of 2g/kg, respectively, by dissolving in drinkable water, according to the following table, to make 6 kinds of corresponding pellet feeds. The experimental design was as follows:
TABLE 5 Water extract anti-Fish stress test design
| Additive material | Blank space | Induction of | |
1 | 2 | 3 | 4 | 5 |
| Aqueous extract of Angelica sinensis | × | × | □ | × | □ | □ | □ | □ |
| Aqueous extract of mulberry leaves | × | × | □ | □ | × | □ | □ | □ |
| Aqueous extract of hawthorn | × | × | □ | □ | □ | × | □ | □ |
| Water extract of wild licorice | × | × | □ | □ | □ | □ | × | □ |
| Aqueous extract of motherwort | × | × | □ | □ | □ | □ | □ | × |
| Stocking density | 0.5 tail/ |
1 tail/ |
1 tail/ |
1 tail/ |
1 tail/ |
1 tail/ |
1 tail/ |
1 tail/L |
Remarking: the basal feed without any water extract is marked as 'blank', the feed with all 5 water extracts is marked as 'control', and the feeds with the water extracts of angelica sinensis, mulberry leaf, hawthorn, wild licorice and motherwort subtracted one by one are respectively marked as '1', '2', '3', '4' and '5' compared with the 'control'.
Selecting 675 tails of crucian fries with the weight of 13.5 +/-0.3 g under the optimal stress density (1.00 tail/L water) and the same culture environment, randomly dividing the crucian fries into 8 treatment groups, and 3 fish tanks in each group. The fish tails of 8 treatment groups were: 15. 30, 30 and 30 tails/cylinder, designated "blank (K)", "induced (Y)", "control (a)" respectively0)”、“1(A1)”、“2(A2)”、“3(A3)”、“4(A4) "and" 5 (A)5) Groups, fed with feed labeled "blank", "control", "1", "2", "3", "4" and "5", respectively. And measuring the growth, the food intake and the feed efficiency of the crucian after 28 days.
As shown in table 6 and fig. 4, the results showed that the growth, food intake and feed efficiency of the induced herds were significantly reduced compared to the blank group; however, feeding the complex water extract restored fish growth, feed intake and feed efficiency to a different extent compared to the induction group. The weight gain recovery rate is in the order of magnitude: 2 > 4 > control > 3 > 1 > 5, 76.92%, 68.26%, 57.23%, 48.69%, 35.08%, 26.79%, respectively. Therefore, in the components of the composite extract, the mulberry leaf water extract and the wild licorice root water extract have an inhibiting effect on the anti-stress effect of other components of the composite extract; the hawthorn water extract, the Chinese angelica water extract and the motherwort water extract play a promoting role. Therefore, the components of the primarily screened compound extract are hawthorn water extract, Chinese angelica water extract and motherwort water extract, and are marked as compound anti-stress agent II.
Table 6 influence of feeding of compound water extract on crucian carp breeding density stress
Remarking: 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). Weight gain is equal to end weight-initial weight; feed efficiency was 100 x gain/feed intake.
3. Natural composite anti-stress agent I and natural composite anti-stress agent II composite anti-stress agent and verification
The components of the compound anti-stress agent I preliminarily screened in the test are wild liquorice ethanol extract, angelica ethyl acetate extract and mulberry leaf ethyl acetate extract. To verify the anti-stress effect, compound anti-stress agent I (abbreviated as "I") was added to the basal diet and compared with feed No. 5 (abbreviated as "control I") having the best effect in component screening test I under the same breeding environment and conditions as above. The test primarily screens the components of the compound anti-stress agent II, which are hawthorn water extract, angelica water extract and motherwort water extract. To verify the anti-stress effect, compound anti-stress agent II (abbreviated as 'II') was added to the basal diet and compared with feed No. 2 (abbreviated as 'control I') which showed the best effect in the component screening test under the same breeding environment and conditions. The feed containing the compound anti-stress agent I (the content of which is 0.3%) and the feed containing the compound anti-stress agent II (the content of which is 0.6%) are mixed in equal amount (the average content is 0.45%) (abbreviated as [ (I + II)/2) ], and are compared with the feed containing the compound anti-stress agent I and the feed containing the compound anti-stress agent II under the same culture environment and conditions. Basal feed was fed to the blank and induction groups as well. The experimental design is shown in table 7 below.
TABLE 7 test design for composite antistress agent
The experimental results after 28 days of feeding are shown in table 8 and figure 5, and compared with the blank group, the growth, food intake and feed efficiency of the induced group fish are significantly reduced; however, feeding the complex anti-stress agent restored fish growth, feed intake and feed efficiency to a different extent than the induction group. The weight gain recovery rate is in the order of magnitude: i > control I; II > control II; (I + II)/2 is more than I and more than II. Therefore, the anti-stress effect of the compound anti-stress agents I and II is higher than that of the control, and the mixed effect of the two agents is better.
TABLE 8 influence of feeding compound anti-stress agent on crucian carp breeding density stress
Remarking: note: 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). Weight gain is equal to end weight-initial weight; feed efficiency was 100 x gain/feed intake.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are merely exemplary embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalent substitutions, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. A natural compound for improving the anti-stress response of fish 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 of wild licorice organic solvent extract, angelica organic solvent extract and mulberry leaf organic solvent extract; the natural compound anti-stress agent II comprises two or more of water extract of fructus crataegi, water extract of radix Angelicae sinensis and water extract of herba Leonuri.
2. The natural composition for improving fish anti-stress response as claimed in claim 1, wherein the organic solvent extract of wild licorice is ethanol extract of wild licorice, the organic solvent extract of angelica is ethyl acetate extract of angelica, and the organic solvent extract of mulberry leaf is ethyl acetate extract of mulberry leaf.
3. The natural compound for improving the anti-stress response of the fish as claimed in claim 1, wherein the natural compound anti-stress agent I comprises three kinds of wild licorice organic solvent extract, Chinese angelica organic solvent extract and mulberry leaf organic solvent extract, and the mass ratio of the wild licorice organic solvent extract to the Chinese angelica organic solvent extract to the mulberry leaf organic solvent extract is (1-2): 1-2 in sequence.
4. The natural compound for improving the anti-stress response of the fish as claimed in claim 1, wherein the natural compound anti-stress agent II comprises three kinds of aqueous extracts of hawthorn, Chinese angelica and motherwort, and the mass ratio of the aqueous extracts of hawthorn, Chinese angelica and motherwort is (1-2) to (1-2) in sequence.
5. The natural compound for improving the 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 complex for increasing the stress resistance of fish according to any of claims 1 to 5, for the preparation of an anti-stress additive or feed for freshwater fish; the stress response includes a high density stress response.
7. A fish feed comprising a basal feed and a natural complex according to 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. A method for preparing a fish feed as claimed in claim 7 or claim 8, wherein the natural complex anti-stress agent I is dissolved in the oil and mixed, and then added to the basic feed material and mixed; and/or dissolving natural compound anti-stress agent II in water, mixing, adding into basic feed raw material, and mixing.
10. A method of preparing a fish feed as claimed in claim 9 wherein the basal feed material is a natural feed.
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