CN110089641B - Motherwort extract, application of motherwort extract in fish feed, fish feed and preparation method of fish feed - Google Patents
Motherwort extract, application of motherwort extract in fish feed, fish feed and preparation method of fish feed Download PDFInfo
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Abstract
The invention discloses a motherwort extract, application of the motherwort extract in fish feed, the fish feed and a preparation method of the fish feed, and solves the problems that the effective components of the conventional motherwort extract used in a fish feed additive are undefined, the extraction rate is low, the growth promoting effect in high-density culture of freshwater fish is not obvious, and the improvement needs to be further carried out according to the action mechanism. The invention comprises the application of the motherwort extract in the high-density freshwater fish culture fish feed as a growth promoter, which can improve the activity of intestinal lipase and amylase, wherein the optimal addition amount of the motherwort extract is 0.35-0.4%. The invention has the advantages of good motherwort extraction effect, good growth promoting effect on the freshwater fish during high-density culture, and the like.
Description
Technical Field
The invention relates to the technical field of fish feed, and particularly relates to a motherwort extract and application thereof, fish feed and a preparation method thereof.
Background
Herba Leonuri (Leonurus artemisinia (Laur.) S.Y.Hu F) is a annual or biennial herb of the genus Leonurus of the family Labiatae. The plants are distributed in most regions in China and live in mountain wastelands, ridges, grasslands and the like. The dry aerial parts of motherwort are the common traditional Chinese medicines. The modern pharmaceutical research result shows that the motherwort is rich in alkaloid, diterpene, flavone, polyphenol, polysaccharide and other substances, and has the effects of tranquilizing, easing pain, inhibiting bacteria, resisting inflammation, resisting anoxia, resisting oxidation, resisting stress, resisting fatigue, enriching blood, promoting blood circulation to remove blood stasis, promoting blood coagulation and improving the yield of laying hens.
Fish farming requires a large amount of feed, and the growth of fish determines the yield of fish, so in the existing aquaculture and feed processing, growth promoters, also called growth promoters, are often added to the feed. The most commonly used growth promoters in feed have traditionally been antibacterial agents (e.g., antibiotics, sulfonamides, furans, and the like) and hormones or hormone analogs. The use of antibiotics has posed a range of hazards to the animals themselves, animal products, and the environment. At present, the use of antibiotics in aquatic feeds is strictly limited in China. Some antibiotics, such as olaquindox, n-triacontanol, and sulfonamides, have been banned. In consideration of the possible harm of hormone to human, the addition of adrenalin receptor agonist, sex hormone, protein assimilation hormone, psychotropic drug, etc. to feed has been prohibited in the regulations on feed and feed additive management issued in 29.5.1999 in China. From the viewpoints of food safety, human health and environmental protection, the development of a novel and environment-friendly growth promoter meets the requirements of sustainable development, is an inevitable direction of feed industry development, and has important practical significance.
Therefore, various plant extracts are researched and used as fish feed additives in the prior art, motherwort is reported as the fish feed additives, but most of the existing methods are that motherwort is directly added into fish feed, and the adding mode cannot well release the active ingredients of motherwort, so that the growth promoting effect on fish is limited.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: the effective components of the existing motherwort herb used for the fish feed additive are undefined and have low utilization rate, and the growth promoting effect in high-density culture of freshwater fish is not obvious and needs to be further improved according to the action mechanism.
The invention provides a motherwort extract for solving the problems, application of the motherwort extract in fish feed, the fish feed and a preparation method of the fish feed.
The invention is realized by the following technical scheme:
the motherwort extract is applied to fish feed, and the motherwort extract is used as a growth promoter in the high-density cultured fish feed of freshwater fish.
A motherwort extract comprises one or more of a motherwort petroleum ether extract, a motherwort ethyl acetate extract, a motherwort ethanol extract and a motherwort water extract, wherein the motherwort petroleum ether extract is obtained by extracting and mixing motherwort with petroleum ether for multiple times, the motherwort ethyl acetate extract is obtained by extracting and mixing filter residue I obtained after the petroleum ether extraction with ethyl acetate for multiple times, the motherwort ethanol extract is obtained by extracting and mixing filter residue II obtained after the ethyl acetate extraction with ethanol for multiple times, and the motherwort water extract is obtained by extracting and mixing filter residue III obtained after the ethanol extraction with water for multiple times.
A herba Leonuri extract comprises one or two of herba Leonuri ethyl acetate extract and herba Leonuri ethanol extract, wherein the herba Leonuri ethyl acetate extract is obtained by extracting residue I after petroleum ether extraction with ethyl acetate for multiple times and mixing; the motherwort ethanol extract is obtained by extracting and combining filter residue II obtained after ethyl acetate extraction by adopting ethanol for multiple times.
The invention preferably selects a motherwort extract, which is a motherwort ethanol extract.
Preferably, the leonurus japonicus ethanol extract is obtained by the following method: firstly, extracting motherwort herb for multiple times by adopting petroleum ether to obtain filter residue I, then extracting the filter residue I for multiple times by using ethyl acetate to obtain filter residue II, finally extracting the filter residue II for multiple times by using ethanol to obtain combined filtrate, and finally distilling the filtrate under reduced pressure to remove the ethanol to obtain the motherwort herb ethanol extract.
Preferably, the content ratio of the flavonoids to the polyphenols in the motherwort extract is 1.35-1.45.
Furthermore, the content ratio of flavonoids to polyphenols in the motherwort herb extract is 1.41.
The fish feed comprises 0.35-0.6% of motherwort extract by weight.
Preferably, the motherwort extract is added in the fish feed in an amount of 0.34-0.36% by weight of the total weight of the feed.
Preferably, the fish feed comprises the following components: 34 to 35 percent of crude protein, 5 to 6.3 percent of crude fat and 0.9 to 1.0 percent of phosphorus.
Further, the crude protein was 34.82%, the crude fat was 5.52%, and the phosphorus was 0.93%.
The preparation method of the fish feed with the motherwort herb extract as the additive comprises the following steps:
step 1: drying herba Leonuri and pulverizing to obtain herba Leonuri powder;
step 2: extracting herba Leonuri powder with petroleum ether for several times to obtain residue I and filtrate I, and distilling the filtrate I under reduced pressure to remove petroleum ether to obtain herba Leonuri petroleum ether extract;
and step 3: extracting the filter residue I for multiple times by using ethyl acetate to obtain a filter residue II and a filtrate II, and distilling the filtrate II under reduced pressure until the weight is constant to remove the ethyl acetate, thereby obtaining a motherwort ethyl acetate extract;
and 4, step 4: extracting the filter residue II with ethanol for multiple times to obtain filter residue III and filtrate III, and distilling the filtrate III under reduced pressure to constant weight to remove ethanol to obtain herba Leonuri ethanol extract;
and 5: extracting the residue III with water for several times to obtain residue IV and filtrate IV, and distilling the filtrate IV under reduced pressure to constant weight to remove water to obtain herba Leonuri water extract.
Step 6: adding the above extracts into fish feed.
Preferably, the petroleum ether extract of leonurus, the ethyl acetate extract of leonurus, the ethanol extract of leonurus and the water extract of leonurus are added to the fish feed, respectively.
Preferably, the specific operation of adding the leonurus petroleum ether extract, the leonurus ethyl acetate or the leonurus ethanol extract into the fish feed is as follows: dissolving herba Leonuri petroleum ether extract, herba Leonuri ethyl acetate or herba Leonuri ethanol extract in fish feed oil, and mixing with other materials of fish feed.
The oil is one or a mixture of more of fish oil, soybean oil, corn oil, rapeseed oil, peanut oil and sunflower seed oil.
Preferably, the oil is corn oil.
Preferably, the fish feed comprises the following raw materials in percentage by weight:
motherwort extract: 0.35 to 0.6 percent
Fish meal: 25 percent of
Bean pulp: 32 percent of
DL-methionine: 0.70 percent
Flour: 37 percent of
Fish oil: 1.5 percent
Corn oil: 1.8 percent
Vitamin additives: 1 percent of
Mineral additive: 1 percent.
Further, the vitamin additive comprises retinol and vitamin D 3 DL-alpha-tocopherol, vitamin K 3 Vitamin B 1 Riboflavin, pyridoxine, cyanocobalamin, D-calcium pantothenate, nicotinic acid, D-biotin, inositol, and folic acid.
Further, the mineral supplement comprises: feSO 4 ·7H 2 O、CuSO 4 ·5H 2 O、ZnSO 4 ·7H 2 O、MnSO 4 ·H 2 O、Na 2 SeO 3 ·5H 2 Og and KI.
The invention has the following beneficial effects:
1. according to the invention, the leonurus extract is added into the fresh water fish feed for high-density culture to serve as a growth promoter, so that the activities of intestinal lipase and amylase of the fish are improved, the Activities of Superoxide Anion (ASA) and hydroxyl radical (AHR) resistance of intestinal tissues are improved, the content of Malondialdehyde (MDA) in the intestinal tissues is reduced, the growth of the fish is facilitated, and the food intake and the feed efficiency are obviously improved.
2. The leonurus extract is leonurus ethanol extract, the content ratio of flavonoids to polyphenols is 1.35-1.45, and the extract containing the two effective components is added into the fish feed according to the concentration of 0.35-0.6%, so that the effects of promoting growth and improving ingestion are remarkable, and under the same growth condition, the culture density of freshwater fish can be improved, the culture yield is improved, and the culture area is reduced.
3. The method adopts a plurality of different solvents to carry out multi-step extraction on the motherwort, the dry extract obtained in each step has different quantities and the contents of flavonoids and polyphenols in different extracts are also different, and the quantity of the flavonoids and the polyphenols extracted by ethanol is the largest after the extraction by petroleum ether and ethyl acetate through screening, the effect is the best, and the extraction effect on the flavonoids is better than that on the polyphenols.
4. According to the invention, by improving the adding mode of the motherwort extract, the extract is firstly added into the grease and then added into other feed raw materials, so that the dispersibility of the motherwort extract in the feed is good, and the feed efficiency is improved.
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 relationship between the cultivation density and the weight gain of crucian carp.
FIG. 2 is a graph showing the relationship between the cultivation density and the feed efficiency.
FIG. 3 is a graph showing the relationship between the cultivation density and the mortality rate of the present invention.
Fig. 4 is a graph showing the relationship between the feeding rate, intestinal lipase and amylase activity and polyphenol content of crucian after feeding motherwort herb extract.
Fig. 5 is a graph showing the relationship between the ingestion rate, intestinal lipase and amylase activity and flavonoid content of crucian after feeding with the motherwort herb extract.
Fig. 6 is a graph showing the relationship between the content of superoxide anion activity (ASA), hydroxyl radical Activity (AHR) and Malondialdehyde (MDA) in intestinal tissues of crucian carp and the content of polyphenols in the intestinal tissues of crucian carp after 10 days of feeding the leonurus extract.
Fig. 7 is a graph showing the relationship between the content of superoxide anion activity (ASA), hydroxyl radical Activity (AHR) and Malondialdehyde (MDA) in intestinal tissues of crucian carp and the content of flavonoids in the intestinal tissues of crucian carp, which are fed with motherwort herb extract for 10 days.
Fig. 8 is a weight gain regression analysis chart of crucian carp after feeding the feed containing ethanol extracts of motherwort with different concentrations for 60 days.
Fig. 9 is a graph of regression analysis of food intake for crucian carp after feeding the feeds containing ethanol extracts of leonurus japonicus at different concentrations for 60 days.
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.
Example 1
1000g of overground part of motherwort is obtained, dried in a forced air drying oven at 50 ℃ to constant weight, crushed and sieved by a 18-mesh sieve to obtain motherwort powder.
Taking 400g of motherwort powder, mixing the motherwort powder with petroleum ether according to the proportion of 1; filtering the mixed solution to obtain clear filtrate and residue, and repeatedly extracting the residue for 3 times to combine the filtrate and obtain residue I; distilling the filtrate under reduced pressure to constant weight, and removing petroleum ether to obtain herba Leonuri petroleum ether extract. Mixing residue I after extraction of leonurus japonicus petroleum ether with ethyl acetate according to the proportion of 1; filtering the mixed solution to obtain clear filtrate and residue, and repeatedly extracting the residue for 3 times to combine the filtrate and obtain residue II; distilling the filtrate under reduced pressure to constant weight, and removing ethyl acetate to obtain herba Leonuri ethyl acetate extract. Mixing residues II after the extraction of the motherwort ethyl acetate with ethanol 1 to 7-9, and stirring for 6 hours at a speed of 800 revolutions per minute; filtering the mixed solution to obtain clear filtrate and residue, and repeatedly extracting the residue for 3 times to combine the filtrate and obtain residue III; distilling the filtrate under reduced pressure to constant weight, and removing ethanol to obtain herba Leonuri ethanol extract; mixing residues III obtained after the ethanol extraction of the motherwort with water in a ratio of 1-9, and stirring for 6 hours at a speed of 800 revolutions per minute; centrifuging the mixed solution to obtain clear supernatant and residue, and repeatedly extracting the residue for 3 times to combine the supernatants; distilling the supernatant under reduced pressure to constant weight, and removing water to obtain herba Leonuri water extract.
The contents of polyphenols and flavonoids in the four motherwort herb extracts were determined by Folin-Ciocalteau reagent method and aluminum nitrate color development method, respectively, and the results are shown in Table 1.
TABLE 1 yield of extract of motherwort, content of flavonoids and polyphenols
Data are presented as mean ± standard deviation of 3 replicates; differences in the upper-case letters in the same column data indicate significant differences (P < 0.05).
The content ratio of flavonoids to polyphenols in the leonurus japonicus petroleum ether extract is 2.73;
in the motherwort ethyl acetate extract, the content ratio of obtained flavonoids to polyphenols is 1.76;
the content ratio of flavonoids to polyphenols in the motherwort ethanol extract is 1.41;
the content ratio of flavonoids to polyphenols in the motherwort water extract is 2.51;
as can be seen from the above table, the dry extracts obtained by different solvent extractions have different yields, which are shown as: the contents of flavonoids and polyphenols in dry extracts obtained by different solvents are different, and the specific values are as follows: the ethanol extract, the ethyl acetate extract, the water extract and the petroleum ether extract show that the content ratio of flavonoids to polyphenols is 1.41-1.76-2.51-2.73, and prove that in the four solvents, after the petroleum ether and the ethyl acetate are sequentially adopted for extraction, the ethanol is used for extracting the flavonoids and the polyphenols to have the best extraction effect, and the extraction effect on the flavonoids is better than that on the polyphenols.
And after the petroleum ether and the ethyl acetate are subjected to early-stage extraction, extracting the filter residue II obtained by the pretreatment by using ethanol as a solvent, wherein the extraction amounts of polyphenols and flavonoids reach the maximum, and are respectively 50.27-55.43g/kg and 71.28-78.00g/kg.
Example 2
In order to explore the influence of the leonurus extract as a fish feed additive on the high-density culture of freshwater fish, the following experiments are carried out:
1. the influence of the change of the breeding density on the fish body weight, the food intake and the feed efficiency is researched.
Purchasing a plurality of crucian fries locally, temporarily culturing for a week in a culturing room, selecting 720 tails of crucian fries with the weight of 8.0 +/-0.2 g, randomly dividing the crucian fries into 8 treatment groups, wherein the number of the tails of each group of 4 fish tanks is as follows: 5. 10, 15, 20, 25, 30, 35 and 40 tails/cylinder. Wherein, the shape, the size and the color of each fish tank are the same, the size is 30cm multiplied by 40cm, and one 800L/h flow aerator is arranged in each fish tank. Tap water is used as a water source, and the volume of water in the tank is 31L. Controlling the culture room temperature to be 22 +/-2 ℃; the oxygen increasing machine continuously supplies oxygen; feeding fish meal and soybean meal type omnivorous fish feed; changing water every 2-3 days, and culturing for 60 days. After a 60-day breeding period, the weight gain, feed intake, feed efficiency and mortality of the treated groups were examined. The influence of the breeding density on the weight and the food intake of the crucian carp is shown in a table 2; the effects of the breeding density on fish weight gain, feed efficiency and mortality are shown in figures 1-3, respectively, where weight gain = final weight-initial weight; feed efficiency =100 x weight gain/feed intake. Data in the figure are presented as mean ± standard deviation of 4 replicates; the difference in the upper-case letters in the same column data indicates significant difference (P < 0.05).
TABLE 2 influence of cultivation Density on weight and food intake of Carassius auratus
Data are expressed as mean ± standard deviation of 4 replicates; in the same index data, the difference of the upper-marked letters indicates that the difference between the data is significant (P < 0.05).
As can be seen from table 2 and fig. 1 to 3, when the stocking density is 0.16-0.48 fish/L water, the crucian gains most weight and within this range, the weight gain is relatively stable; the weight gain gradually decreases along with the increase of the culture density, and the culture density with the minimum weight gain is 0.97-1.29 tails per L of water. The maximum feeding density is 0.32-0.48 tails per L of water; with the increase of the culture density, the food intake is gradually reduced, and the minimum density is 0.97-1.29 tail/L water. The stocking density with the highest feed efficiency is 0.16-0.48 per liter of water; with the increase of the breeding density, the feed efficiency is gradually reduced, and the density with the lowest feed efficiency is 0.97-1.29 tail/L water. When the stocking density is 1.13-1.29 tail/L water, the crucian death occurs. When the breeding density is 0.48 fish/L water, the weight gain, food intake and feed efficiency of the crucian are highest, and the optimal breeding density is determined to be 0.48 fish/L water. With the increase of the density, the weight gain, the food intake and the feed efficiency of the crucian gradually decrease; when the density of the crucian increases to 0.97 tail/L water, the weight gain, the food intake and the feed efficiency of the crucian are reduced to the lowest level; when the density of the crucian carp is further improved to 1.13 fish/L water, the crucian carp dies. Comparing under the condition of the above culture density gradient, obtaining the highest culture density of water with 0.97 tail/L which does not cause death of the crucian.
2. Research on influence of leonurus extract in different extraction steps on growth of crucian in high-density culture
Firstly, 4 motherwort herb extracts are obtained according to the extraction method of example 1;
the second step is that: 4 extracts of motherwort were added to the feed raw materials at concentrations of 0 and 2g/kg feed and mixed according to the fish feed formulation of table 3 below to make a pellet feed with a diameter of 2 mm.
TABLE 3 Fish basic feed formulation
1 The vitamin supplement comprises per kilogram: retinol (500,000IU/g) 0.80g, vitamin D 3 (500,000IU/g) 0.48g, DL-alpha-tocopherol (50%) 20.00g, vitamin K 3 (23%) 0.43g, vitamin B 1 0.11g (90%), 0.63g of riboflavin (80%), 0.92g of pyridoxine (81%), 0.10g of cyanocobalamin (1%), 2.73g of D-calcium pantothenate (90%), 2.82g of nicotinic acid (99%), 5.00g of D-biotin (2%), 52.33g of inositol (99%) and 0.52g of folic acid (96%).
2 The mineral supplement comprises per kilogram: feSO 4 ·7H 2 69.70g of O (containing 20 percent of Fe) and CuSO 4 ·5H 2 1.20g of O (containing Cu 25%), znSO 4 ·7H 2 21.64g of O (containing 23% of Zn) and MnSO 4 ·H 2 4.09g of O (containing Mn 32%), and Na 2 SeO 3 ·5H 2 O (containing Se 1%) 2.50g and KI (containing I4%) 2.90g.
The specific preparation process comprises the following steps:
(1) Calculating the mass of fish oil and corn oil required in unit mass of feed according to the formula of the basic feed, weighing the fish oil and the corn oil according to the mass, and mixing the fish oil and the corn oil.
(2) Calculating the mass of the herba Leonuri extract required in the feed with the same unit mass according to the content of the extract in the designed feed, weighing the herba Leonuri extract, mixing the herba Leonuri extract with the above mixed oil, and ultrasonically oscillating for 20min to mix;
(3) And (3) adding the mixture obtained in the step (2) into the other raw materials of the feed with the same unit mass, and uniformly mixing to prepare the granular feed with the diameter of 2 mm.
The third step: the highest culture density of 0.97 tail/L water which does not cause death of the crucian is selected, and the crucian is fed by adopting the fish feed and the effect is tracked.
Under the same culture environment conditions, 495 crucian fries with the weight of 22.5 +/-0.9 g are selected and randomly divided into 6 treatment groups, and each group has 3 fish tanks. The tail numbers of the crucian carps in the 6 treatment groups are respectively as follows: 15. 30, 30 and 30 tails/jar, respectively designated blank, control, leonurus petroleum ether extract, leonurus ethyl acetate extract, leonurus ethanol extract and leonurus water extract group. The 6 treatment groups are respectively fed with feed containing 0,0 and 2g/kg of leonurus petroleum ether extract, leonurus ethyl acetate extract, leonurus ethanol extract and leonurus water extract. Feeding 4 times per day, 8 in the morning: 00 feeding for the first time, feeding every 4 hours later, 8:00 the last feeding is carried out. Feeding the feed on the 1 st day, wherein each time is 0.5 g/jar; then, 0.1 g/cylinder is added for each feeding; until the feed amount is not increased any more by the residual materials. Later, the residual feed must be ensured every day, the residual feed is quickly fished out 30 minutes after each feeding, the residual feed is dried in time, and the weight is recorded; and increasing the feeding amount after the rest materials disappear.
After the feeding experiment is carried out for 10 days, the food intake and the fish weight of each treatment group are counted, and the food intake rate is calculated. After 1 day of stopping feeding, intestinal samples of the fish were collected, and contents of lipase, amylase, superoxide anion (ASA) and hydroxyl radical resistant Activity (AHR) and Malondialdehyde (MDA) in intestinal tissues of 6 treatment groups were determined by a kit (Nanjing institute of bioengineering).
The detection results are shown in tables 4 and 5, and the results show that feeding four motherwort herb extracts is beneficial to improving the food intake of the fish and the activity and the antioxidation of intestinal digestive enzyme; and the effect is more obvious along with the increase of the contents of flavonoids and polyphenols, wherein the fish ingestion rate, the intestinal lipase and amylase activity, the ASA and AHR are highest, the MDA content is lowest in the leonurus ethanol extract treatment group, and the effect is best.
The results of the correlation analysis are shown in FIGS. 4, 5, 6 and 7, where the data are presented as the mean of 3 replicates. The results show that the leonurus extract has close relation with the contents of polyphenols and flavonoids thereof in improving the ingestion, intestinal digestive enzyme activity and antioxidation effect of the fish. Because polyphenols and flavonoids in the motherwort herb extract improve the intestinal oxidation resistance of the fish, the digestion function of the fish is improved, and the ingestion rate of the fish is further improved.
TABLE 4 ingestion rate of crucian carp, intestinal lipase and amylase activity in 10 days after feeding motherwort herb extract under high density cultivation condition
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).
TABLE 5 intestinal anti-superoxide anion activity (ASA), anti-hydroxy radical Activity (AHR) and Malondialdehyde (MDA) content of Carassius auratus 10 days after feeding with herba Leonuri extract under high density culture conditions
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).
Therefore, although the same amount of leonurus petroleum ether extract, leonurus ethyl acetate extract, leonurus ethanol extract and leonurus water extract is added, the content ratio of flavonoids and polyphenols in the obtained extract is different due to different amounts of flavonoids and polyphenols extracted by different solvents, and finally the growth influence on crucian under the high-density culture condition is different. The concrete expression is as follows: the leonurus japonicus ethanol extract has the largest content of flavonoid and polyphenol, the content ratio of the flavonoid to the polyphenol is 1.41, the digestion function and the ingestion rate of crucian are improved to the greatest extent, and the growth promoting effect is the best.
3. Research on influence of different addition amounts of leonurus japonicus ethanol extract in fish feed on growth of crucian
Because the content of flavonoids and polyphenols in the leonurus japonicus ethanol extract is highest, the content ratio of flavonoids to polyphenols is 1.41, the effect is best, the extract is selected as a fish feed additive, the leonurus japonicus ethanol extract is added into feed according to the concentration of 0, 1, 2, 3, 4, 5 and 6g/kg respectively according to the third step method to prepare corresponding 7 kinds of pellet feed, the tail of the crucian carp fry 900 with the weight of 6.40 +/-0.19 g is selected under the conditions of the stocking density of 0.97 tail/L water and the same breeding environment, the treatment groups are randomly divided into 8 treatment groups, and each group has 4 fish tanks. The fish tails of 8 treatment groups were: 15. 30, 30 and 30 tails/cylinder, designated blank, control, 1g, 2g, 3g, 4g, 5g and 6g groups, respectively. The 8 treatment groups are fed with feed containing 0, 1, 2, 3, 4, 5 and 6g/kg of motherwort ethanol extract respectively. After 60 days, the influence of feeding the leonurus japonicus ethanol extract on the growth, food intake and feed efficiency of crucian carp is measured and broken line regression analysis is carried out.
The results are shown in Table 6, FIGS. 8 and 9, where the data are presented as mean. + -. Standard deviation of 4 replicates.
TABLE 6 influence of feeding motherwort Ethanol Extract (EE) for 60 days on crucian growth, food intake and feed efficiency under high-density culture conditions
Data are presented as mean ± standard deviation of 4 replicates; the difference in the upper-case letters in the same column data indicates significant difference (P < 0.05). Weight gain = end weight-initial weight; feed efficiency =100 x weight gain/food intake.
The results show that the leonurus ethanol extract improves the weight gain, food intake and feed efficiency of crucian, and the weight gain, food intake and feed efficiency are gradually improved along with the increase of the addition of the leonurus ethanol extract, and by adding the leonurus ethanol extracts with different concentrations into the fish feed, compared with a control group, the weight gain rate is increased by 43-133 percentage points, the food intake is increased by 11.4-33.6 percentage points, and the feed efficiency is increased by 6-17.5 percentage points. However, when the addition amount is increased to a certain amount, the increase of weight gain, food consumption and feed efficiency is greatly reduced, and regression analysis shows that the addition amount of the leonurus japonicus ethanol extract is 3.49g/kg as the optimal addition amount from the viewpoint of growth promoting effect and economy, and the addition amount of the leonurus japonicus ethanol extract is 3.55g/kg as the optimal addition amount from the viewpoint of food consumption and economy, so that the optimal addition amount of the leonurus japonicus ethanol extract in the fish feed is preferably 3.49-3.55g/kg, and the comprehensive effect on crucian culture is best.
The invention improves the problems of slow growth and reduced ingestion rate of crucian caused by high-density culture, and can improve the culture density under the same crucian growth condition after adding the leonurus japonicus ethanol extract, thereby being beneficial to improving the culture yield and reducing the culture area.
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 (4)
1. The application of the leonurus ethanol extract in the fish feed is characterized in that the leonurus ethanol extract is used as a growth promoter in the high-density culture fish feed of freshwater fish;
the preparation method of the motherwort ethanol extract comprises the following steps:
step 1: drying herba Leonuri and pulverizing to obtain herba Leonuri powder;
and 2, step: taking 400g of motherwort powder, mixing the motherwort powder with petroleum ether according to the proportion of 1; filtering the mixed solution to obtain clear filtrate and filter residue, repeatedly extracting the filter residue for 3 times, and mixing the filtrates to obtain filter residue I and filtrate I; distilling the filtrate I under reduced pressure until the weight of the filtrate is constant, and removing petroleum ether to obtain a leonurus petroleum ether extract;
and step 3: mixing the filter residue I extracted from the leonurus japonicus by using the petroleum ether with ethyl acetate according to the proportion of 1; filtering the mixed solution to obtain clear filtrate and filter residue, repeatedly extracting the filter residue for 3 times, and mixing the filtrates to obtain filter residue II and filtrate II; distilling the filtrate II under reduced pressure to constant weight, and removing ethyl acetate to obtain herba Leonuri ethyl acetate extract;
and 4, step 4: mixing the filter residue II extracted from the motherwort ethyl acetate with ethanol 1 at a ratio of 7-9, and stirring for 6 hours at a speed of 800 rpm; filtering the mixed solution to obtain clear filtrate and filter residue, repeatedly extracting the filter residue for 3 times, and mixing the filtrates to obtain filter residue III and filtrate III; distilling the filtrate III under reduced pressure to constant weight, and removing ethanol to obtain herba Leonuri ethanol extract;
the addition amount of the leonurus japonicus ethanol extract accounts for 0.35 to 0.6 percent of the total weight of the feed.
2. The use of the ethanol extract of leonurus japonicus as claimed in claim 1 in fish feed, wherein the fish feed comprises the following components: 34 to 35 percent of crude protein, 5 to 6.3 percent of crude fat and 0.9 to 1.0 percent of phosphorus.
3. The application of the leonurus ethanol extract in the fish feed according to claim 1, wherein the specific operation of adding the leonurus ethanol extract into the fish feed is as follows: dissolving the leonurus ethanol extract in oil of the fish feed, and mixing the oil with the rest materials of the fish feed, wherein the oil is one or a mixture of more of fish oil, soybean oil, corn oil, rapeseed oil, peanut oil and sunflower seed oil.
4. The application of the leonurus japonicus ethanol extract in the fish feed as claimed in claim 1 or 3, wherein the fish feed comprises the following raw materials in percentage by weight:
fish meal: 25 percent
Bean pulp: 32 percent of
DL-methionine: 0.70 percent
Flour: 37 percent
Fish oil: 1.5 percent
Corn oil: 1.8 percent
Vitamin additives: 1 percent
Mineral additive: 1 percent.
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