CN109965148A - Mulberry-leaf extract inhibit freshwater fish culturing stress aspect application and feed and feed producing method - Google Patents
Mulberry-leaf extract inhibit freshwater fish culturing stress aspect application and feed and feed producing method Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The present invention provides a kind of mulberry-leaf extract inhibit freshwater fish culturing stress aspect application and feed and feed producing method.Mulberry-leaf extract the preparation method comprises the following steps: by mulberry leaf dries pulverizing, hexamethylene is added and extracts, uses extracted by ether after filtering again, refilters, filtrate decompression is distilled to constant weight and is made.The present invention also provides the preparation methods of this mulberry-leaf extract proportion and feed in anti-stress feed.The present invention applies mulberry-leaf extract as Antistress agent in feed, it can inhibit fresh-water fishes in cultivation because of stress reaction caused by cultivating high density, copper sulphate and metrifonate etc., to improve the anti-stress ability of fresh-water fishes, reduce its morbidity and mortality, mulberry leaf new application is developed simultaneously, is effectively utilized mulberry leaf resource.
Description
Technical field
The invention belongs to fish culture technical fields, and in particular to a kind of mulberry-leaf extract is inhibiting the freshwater fish culturing stress
The application of aspect and feed and feed producing method.
Background technique
It is the important feature of current aquaculture that high density is intensive.High-density breeding improves feeding amount, supports in water body
It grows waste to increase, makes water degradation, show as dissolved oxygen in water reduction, ammonia nitrogen and content of nitrite are exceeded.It has been reported aobvious
Show: these factors will lead to the oxidativestress damage of fish, and then reduce digestion and absorption of the fish to feed, reduces the growth of fish and exempts from
Epidemic disease function improves the morbidity and mortality of fish.High-density breeding also leads to the Use out of range of chemosterilant and insecticide.Sulphur
Sour copper is one of metal class disinfection pesticides most-often used in aquaculture, and effective use concentration in water is generally
0.5~1.0mg/L.Existing research confirms that breeding water body can induce the oxidative stress of fish using copper sulphate and then reduce taking the photograph for fish
Food, digestion and absorption, growth and immune function.Metrifonate is one of organophosphorus insecticides most-often used in aquaculture, is
A kind of anticholinesterase.Metrifonate poisoning can cause animal muscle in disorder, and disequilibrium.Display has been reported,
Metrifonate can induce cultured fishes and oxidative stress occurs and adversely affects to the digestion and absorption, breathing and immune function of fish.
When stimulation of the fish by these stress factors, adrenal cortex is loose, the water such as internal glucocorticoid and steroid cortin
It is flat to rise, thus cause the activity of blood lymphocyte, macrophage and neutrophil cell and lysozyme sharply to decline, animal
Immune system is suppressed, and performance weight gain slows down, and the death rate increases, to seriously affect the economic benefit and Ecological Effect of cultivation
Benefit.
Mulberry tree is planted in that China is with a long history, and mulberry leaf are mainly used to breed silkworms, also edible and be used as medicine.Modern pharmacy research knot
Fruit shows that mulberry leaf have a variety of pharmacological activity, has the function of removing In vitro and in vivo activity oxygen (ROS) and chelated metal ions, but
At present related mulberry-leaf extract inhibit freshwater fish culturing stress aspect using having not been reported.China's mulberry leaf resource is abundant, but
Using extremely limited, there are a large amount of mulberry leaf to be discarded every year.Mulberry leaf new application is developed, improves traditional mulberry leaf using structure, plays
The Potential Comprehensive Utilization of mulberry leaf, turns waste into wealth, and improves mulberry leaf economic and social benefit, has a very important significance.
Summary of the invention
For the above-mentioned problems in the prior art, the present invention provides a kind of mulberry-leaf extracts, and fresh-water fishes to be inhibited to support
Grow stress aspect application and feed and feed producing method.It is applied mulberry-leaf extract as Antistress agent in feed, energy
Inhibit freshwater fish culturing stress reaction, reduces the influence of the stress factors such as cultivation high density, copper sulphate and metrifonate, improve fresh water
The anti-stress ability of fish reduces its morbidity and mortality, while developing mulberry leaf new application, is effectively utilized mulberry leaf resource.
To achieve the above object, the technical solution adopted by the present invention to solve the technical problems is: the present invention provides mulberries
Leaf extract inhibit freshwater fish culturing stress aspect application.
Further, mulberry-leaf extract is added in feed and is used, adding proportion is 4.52~7.00wt ‰.
Further, mulberry-leaf extract is mulberry leaf ether extract, mulberry leaf hexamethylene extract or mulberry leaf acetone extract.
Further, mulberry leaf hexamethylene extract is made by the following method: by mulberry leaf drying, crushing, Mulberry Leaf and ring
Hexane is mixed in the ratio of 1:5~8, and stirring is filtered after extracting, and is repeated to extract 2~3 times, is obtained filtrate and the first filter residue, merging filtrate
After be evaporated under reduced pressure to constant weight, be made；
Mulberry leaf ether extract is made by the following method: will be first obtained in mulberry leaf hexamethylene extract extraction process
Filter residue and ether are mixed in the ratio of 1:5~8, and stirring is filtered after extracting, and repeat to extract 2~3 times, obtain no residue filtrate and the second filter
Slag, vacuum distillation is made to constant weight after merging filtrate；
Mulberry leaf acetone extract is made by the following method: by the second filter obtained in mulberry leaf ether extract extraction process
Slag and acetone are mixed in the ratio of 1:5~8, and stirring is filtered after extracting, and repeat to extract 2~3 times, obtain no residue filtrate and third filter
Slag, vacuum distillation is made to constant weight after merging filtrate.
A kind of fresh-water fishes Antistress agent, including above-mentioned mulberry-leaf extract.
A kind of feed, including basal feed and above-mentioned mulberry-leaf extract.
Further, mulberry-leaf extract accounts for 4.52~the 7.00 ‰ of feed total weight.
Further, mulberry-leaf extract accounts for the 4.90 ‰ of feed total weight.
Further, basal feed includes following parts by weight of component: 23.00~25.00 parts of fish meal, dregs of beans 31.00~33.00
Part, 0.60~0.80 part of DL- METHIONINE, 36.00~38.00 parts of flour, 3.00~3.50 parts of grease, vitamine supplement 0.80
~1.30 parts and 0.80~1.30 part of mineral additive.
Wherein, vitamine supplement includes following parts by weight of component: 0.80 part of retinol, vitamin D30.48 part, DL-
20.00 parts of alpha-tocopherol, vitamin K30.43 part, vitamin B10.11 part, 0.63 part of riboflavin, 0.92 part of pyridoxol, cyanogen cobalt
0.10 part of amine, 2.73 parts of D-VB5 calcium, 2.82 parts of niacin, 5.00 parts of D-Biotin, 52.33 parts of inositol and 0.52 part of folic acid.
Mineral additive includes following parts by weight of component: FeSO4·7H269.70 parts of O, CuSO4·5H21.20 parts of O,
ZnSO4·7H221.64 parts of O, MnSO4·H24.09 parts of O, Na2SeO3·5H22.50 parts and 2.90 parts of KI of O.
Grease is the mixture that soybean oil, corn oil, rapeseed oil, peanut oil or sunflower oil are mixed with fish oil.
The preparation method of above-mentioned feed, comprising the following steps:
Mulberry-leaf extract is dissolved in grease, is then mixed with remaining component of basal feed, is pelletized, is made.
In conclusion the invention has the following advantages that
1. the present invention in fish meal add mulberry-leaf extract be used as Antistress agent, inhibit cultivate high density, copper sulphate and
The stress reaction that metrifonate etc. generates fresh-water fishes improves the anti-stress ability of fresh-water fishes, inhibits fish body rollover, makes its food ration
Restore with weight gain, and then improves the economic benefit of freshwater fish culturing.
2. present invention determine that optimum addition of the mulberry leaf ether extract in anti-stress feed is 4.90g/kg.Its
In, anti-cultivation high density, sulfuric-resisting copper and anti-metrifonate stress the optimal mulberry leaf ether extract concentration of effect be respectively
4.85g/kg, 4.52g/kg and 4.90g/kg feed.
3. using two steps the present invention provides the extracting method of mulberry-leaf extract and extracting, first carried out just using hexamethylene
Step is extracted, and is reused ether and is further extracted, and mulberry leaf ether extract is obtained；Greatly reduce the ring in mulberry leaf ether extract
Hexane extract component, extracts active constituent purity is high, and extraction effect is good.
4. present invention determine that in mulberry-leaf extract anti-freshwater fish culturing stress active constituent be flavone compound.Mulberry
The chelated metal ions (MCA) of leaf extract, restore fresh-water fishes ingest and improve fish enteron aisle anti-superoxide activities (ASA) and
There are close with its Flavonoid substances content for the effect of anti-hydroxy radical active (AHR) and reduction fish enteron aisle malonaldehyde (MDA) content
Cut relationship.The mechanism of the anti-high density stress of mulberry-leaf extract is that it improves body anti-oxidation function.Wherein, mulberry leaf extracted by ether
The flavonoid content highest of object is 61.68~65.70g/kg.
5. passing through the present invention provides the method in feed is added to using mulberry-leaf extract as fresh-water fishes Antistress agent
It is dissolved into grease, then obtains fish meal with other raw material mixing granulations again；By extra mulberry leaf resource change give up into
Treasured is effectively utilized mulberry leaf resource abundant, enriches the purposes of mulberry leaf, improves economic benefit.
Detailed description of the invention
Fig. 1 is the external chelated metal ions of opposed polarity mulberry-leaf extract active (MCA) and feeds rear crucian and ingest recovery
Rate schematic diagram；
Fig. 2 ingests for opposed polarity mulberry-leaf extract Flavone content with chelated metal ions active (MCA) and crucian extensive
Multiple rate relation schematic diagram；
Fig. 3 is the influence schematic diagram that cultivation density increases weight to crucian；
Fig. 4 is influence schematic diagram of the cultivation density to crucian carp feed efficiency；
Fig. 5 is influence schematic diagram of the cultivation density to the crucian death rate；
Fig. 6 is crucian enteron aisle anti-superoxide activities (ASA), the anti-hydroxy radical for feeding opposed polarity mulberry-leaf extract
The schematic diagram of active (AHR) and malonaldehyde (MDA) content；
Fig. 7 is each extract flavone class content of mulberry leaf and crucian enteron aisle anti-superoxide activities (ASA), anti-hydroxy radical
The relational graph of active (AHR) and malonaldehyde (MDA) content；
Fig. 8 is to feed mulberry leaf ether extract 60 days, crucian carp weight gain recovery rate broken line regression analysis schematic diagram；
Fig. 9 is influence schematic diagram of the different sulfuric acid copper contents to crucian grazing rate in water；
Figure 10 is influence schematic diagram of the different sulfuric acid copper contents to the crucian death rate in water；
Figure 11 is metrifonate contents different in water to the influence schematic diagram of crucian rollover rate；
Figure 12 is influence schematic diagram of the different metrifonate contents to the crucian death rate in water；
Figure 13 be copper sulphate stress rear crucian carp ingest recovery rate broken line regression analysis schematic diagram；
Figure 14 is that metrifonate stress rear crucian carp rollover inhibiting rate broken line regression analysis schematic diagram.
Specific embodiment
Embodiment 1
Mulberry-leaf extract, preparation method are as follows:
(1) 1000g mulberry leaf are dried in 50 DEG C of air dry ovens to constant weight, crushed 18 meshes, obtain Mulberry Leaf；
(2) Mulberry Leaf 400g is taken, is mixed in the ratio of Mulberry Leaf and hexamethylene 1:6g/mL, 600r/min stirs 6h,
Clear filtrate and filter residue are obtained after filtering；
(3) filter residue is repeated into extraction 2 times by step (2), merges the filtrate of 3 extractions, filtrate decompression is distilled to perseverance
Hexamethylene is removed again, obtains mulberry leaf hexamethylene extract.
Embodiment 2
Mulberry-leaf extract, preparation method are as follows:
(1) 1000g mulberry leaf are dried in 50 DEG C of air dry ovens to constant weight, crushed 18 meshes, obtain Mulberry Leaf；
(2) Mulberry Leaf 400g is taken, is mixed in the ratio of Mulberry Leaf and hexamethylene 1:6g/mL, 600r/min stirs 6h,
Filter residue is obtained after filtering, and filter residue is repeated into extraction 2 times；
(3) filter residue obtained by step (2) and ether are mixed in the ratio of 1:6g/mL, 600r/min stirs 6h, filters clear
Filter residue is repeated extraction 2 times, merges the filtrate of 3 extractions by clear filtrate and filter residue；
(4) filtrate decompression obtained by step (3) is distilled to constant weight and removes ether, obtain mulberry leaf ether extract.
Embodiment 3
Mulberry-leaf extract, preparation method are as follows:
(1) 1000g mulberry leaf are dried in 50 DEG C of air dry ovens to constant weight, crushed 18 meshes, obtain Mulberry Leaf；
(2) Mulberry Leaf 400g is taken, is mixed in the ratio of Mulberry Leaf and hexamethylene 1:6g/mL, 600r/min stirs 6h,
Filter residue is obtained after filtering, and filter residue is repeated into extraction 2 times；
(3) filter residue obtained by step (2) and ether are mixed in the ratio of 1:6g/mL, 600r/min stirs 6h, obtains after filtering
Filter residue is obtained, filter residue is repeated into extraction 2 times；
(4) filter residue obtained by step (3) and acetone are mixed in the ratio of 1:6g/mL, 600r/min stirs 6h, then will mix
Liquid filtering is closed, clarification is obtained without residue filtrate and filter residue, filter residue is repeated into extraction 2 times, merges the filtrate of 3 extractions；
(5) filtrate decompression obtained by step (4) is distilled to constant weight and removes acetone, obtain mulberry leaf acetone extract.
Embodiment 4
Mulberry-leaf extract, preparation method are as follows:
(1) 1000g mulberry leaf are dried in 50 DEG C of air dry ovens to constant weight, crushed 18 meshes, obtain Mulberry Leaf；
(2) Mulberry Leaf 400g is taken, is mixed in the ratio of Mulberry Leaf and hexamethylene 1:6g/mL, 600r/min stirs 6h,
Filter residue is obtained after filtering, and filter residue is repeated into extraction 2 times；
(3) filter residue obtained by step (2) and ether are mixed in the ratio of 1:6g/mL, 600r/min stirs 6h, filters to obtain filter
Filter residue is repeated extraction 2 times by slag；
(4) filter residue obtained by step (3) and acetone are mixed in the ratio of 1:6g/mL, 600r/min stirs 6h, filters to obtain filter
Filter residue is repeated extraction 2 times by slag；
(5) filter residue obtained by step (4) and water are mixed in the ratio of 1:6g/mL, 600r/min stirs 6h, is centrifuged to obtain clarification
Filter residue is repeated extraction 2 times, merges the filtrate of 3 extractions by filtrate and filter residue；
(6) filtrate decompression obtained by step (5) is distilled to constant weight and removes water, obtain mulberry leaf water extract.
Using the flavonoid content and chelating gold of four kinds of mulberry-leaf extracts obtained by chemical determination Examples 1 to 4
Belong to ion (Fe2+) active (MCA), the result is shown in table 1 and Fig. 1.Meanwhile the MCA of four kinds of mulberry-leaf extracts and its flavonoids are contained
Amount carries out correlation analysis, and analysis result is shown in Fig. 2.Table 1 and Fig. 1 data are indicated with 3 duplicate average ± standard deviations；?
In same index, the female difference person of data shoulder marking-up indicates significant difference between data, and identical person indicates that difference is not significant (P < 0.05).
Fig. 2 data are indicated with 3 duplicate averages.
Flavonoid content in 1 opposed polarity mulberry-leaf extract of table
As shown in Table 1, the Flavone content highest of mulberry leaf ether extract is 61.68~65.70g/kg.It can by Fig. 1
Know, mulberry-leaf extract has the function of chelating transition metal ion, and wherein best with mulberry leaf ether extract effect.By Fig. 2
It is found that the MCA of mulberry-leaf extract is with its Flavonoid substances content, there are substantial connections.Therefore, it is flavones in mulberry-leaf extract
Substance has chelated transition metal ion.
Embodiment 5
Several carp fries are bought in locality, is temporarily supported after a week in culturing chamber, selects 8.0 ± 0.2g of weight carp fry 720
Tail, is randomly divided into 8 processing groups, every group of 4 fish jars, and the fish tail number of 8 processing groups is respectively as follows: 5,10,15,20,25,30,35
With 40 tails/cylinder.Wherein, the form, size of each fish jar are identical with color, and size is all 30cm × 30cm × 40cm simultaneously built-in
One, 800L/h flow aerator.It is 31L by water source, cylinder inner volume of tap water.22 ± 2 DEG C of room temperature of control cultivation；Oxygenation
Machine persistently supplies oxygen；Feed fish meal bean pulp type omnivorefish feed；It is primary to change within every 2~3 days water, the cultivation phase is 60 days.It cultivates within 60 days
After phase, each processing group fish weight gain, food ration, feed efficiency and the death rate are investigated, cultivation density is to crucian weight and food ration
Influence is shown in Table 2, and cultivation density increases weight to fish, Fig. 3~5 are shown in the influence of feed efficiency and the death rate respectively.Wherein, table 2 and Fig. 3~
Data are indicated in 5 with 4 duplicate average ± standard deviations；In same achievement data, Superscript letters difference person indicates data
Between significant difference, identical person indicates that difference is not significant (P < 0.05).Weight gain=end weight-initial weight；Feed efficiency=100 × weight gain/
Food ration.
Influence of 2 cultivation density of table to crucian weight and food ration
As shown in Table 2, with the raising of cultivation density, food ration is in first to increase to subtract trend afterwards, most cultivation density of ingesting
For 0.32~0.48 tail/L water.From the figure 3, it may be seen that weight gain is gradually reduced with the raising of cultivation density, least cultivation of increasing weight is close
Degree is 0.97~1.29 tail/L water.As shown in Figure 4, with the raising of cultivation density, feed efficiency is gradually reduced, and feed efficiency is most
Low density is 0.97~1.29 tail/L water, up to 0.16~0.48 tail/L water.As shown in Figure 5, cultivation density 1.13~
When 1.29 tails/L water, fish starts to occur dead.
From the foregoing, it will be observed that best cultivation density is 0.48 tail/L water, generating maximal density stress be without causing fish is dead to support
Growing density is 0.97 tail/L water.
Embodiment 6
A kind of anti-stress feed, including following parts by weight of component: 24.00 parts of fish meal, 32.00 parts of dregs of beans, DL- METHIONINE
0.70 part, 37.00 parts of flour, 1.50 parts of fish oil, 1.80 parts of sunflower oil, 0.49 part of mulberry-leaf extract, vitamine supplement
1.00 parts and 1.00 parts of mineral additive.
Wherein, vitamine supplement includes following parts by weight of component: 0.80 part of retinol, vitamin D30.48 part, DL-
20.00 parts of alpha-tocopherol, vitamin K30.43 part, vitamin B10.11 part, 0.63 part of riboflavin, 0.92 part of pyridoxol, cyanogen cobalt
0.10 part of amine, 2.73 parts of D-VB5 calcium, 2.82 parts of niacin, 5.00 parts of D-Biotin, 52.33 parts of inositol and 0.52 part of folic acid.
Mineral additive includes following parts by weight of component: FeSO4·7H269.70 parts of O, CuSO4·5H21.20 parts of O,
ZnSO4·7H221.64 parts of O, MnSO4·H24.09 parts of O, Na2SeO3·5H22.50 parts and 2.90 parts of KI of O.
The preparation method of above-mentioned anti-stress feed the following steps are included:
(1) mulberry-leaf extract is first mixed with sunflower oil and fish oil according to feed formula, sonic oscillation 40min keeps it mixed
It is even；
(2) remaining component raw material is added into mixture obtained by step (1), after mixing, the graininess that diameter 2mm is made is raised
Material.
Embodiment 7
4 kinds of extracts of mulberry leaf made from Examples 1 to 4 are added in feed by the concentration of 0 and 2g/kg, according to implementation
Corresponding 5 kinds of pellets are made in formula and preparation method in example 6.
Most preferably stress under density (0.97 tail/L water) and the identical breeding environment condition of embodiment 5, select weight 23.3 ±
495 tail of 1.0g carp fry is randomly divided into 6 processing groups, every group of 3 fish jars.The fish tail number of 6 processing groups is respectively as follows: 15,30,
30,30,30 and 30 tails/cylinder, it is respectively designated as control, induction, mulberry leaf hexamethylene extract, mulberry leaf ether extract, mulberry leaf third
Ketone extract and mulberry leaf water extract group.6 processing groups are fed respectively containing 0,0 and 2g/kg mulberry leaf hexamethylene extract, mulberry leaf second
The feed of ether extract, mulberry leaf acetone extract and mulberry leaf water extract.It feeding 4 times daily, morning, 8:00 fed first time, with
It was fed afterwards every 4 hours once, carry out last time and feed in the evening 8:00.Feed the 1st day, every time 0.5 gram/cylinder；Later every time
It feeds and increases by 0.1 gram/cylinder；Until surplus material occur is not further added by feeding volume.It must assure that surplus material daily later, and thrown in each
Start to pull residual bait out rapidly within 30 minutes after feeding, dries residual bait in time and record weight；Feeding volume is further added by after disappearing etc. surplus material.It throws
After feeding experiment progress 10 days, the food ration of each processing group and the weight of fish are counted, calculates grazing rate.Stop after feeding 1 day, acquires fish
Intestinal samples, 6 processing group intestinal tissues anti-super oxygen yin is measured using kit (Bioengineering Research Institute is built up in Nanjing) method
Active (AHR) and malonaldehyde (MDA) content of ion activity (ASA), anti-hydroxy radical.
The result is shown in Figure 1 and 6, and data are indicated in figure with 3 duplicate average ± standard deviations；In same index, number
Indicate that difference is not significant (P < 0.05) according to significant difference, identical person between the female difference person's expression data of shoulder marking-up.As seen from the figure, mulberry leaf
Object is taken to restore crucian grazing rate and its enteron aisle antioxidant activity；The ether extract effect of mulberry leaf is best.Correlation analysis knot
Fruit sees Fig. 2 and 7, and data are indicated in figure with 3 duplicate averages.As seen from the figure, the recovery of four kinds of extracts of mulberry leaf ingest and
There are substantial connections with its Flavone content for antioxidation；It is due to its chelated metal ions and raising that it, which restores ingestion function,
Enteron aisle oxidation resistance.
Embodiment 8
Mulberry leaf ether extract made from embodiment 2 is added to feeding by the concentration of 0,1,2,3,4,5,6 and 7g/kg respectively
In material, according to the formula and preparation method in embodiment 6, corresponding feed is made.Most preferably stress 0.97 tail of density/L water and
Under the identical breeding environment of embodiment 5,7.2 ± 0.2g of weight carp fry, 1020 tail is selected, is randomly divided into 9 processing groups, every group 4
A fish jar.The fish tail number of 9 processing groups is respectively as follows: 15,30,30,30,30,30,30,30 and 30 tails/cylinder, is respectively designated as pair
According to (D), induction (Y), 1g (E1)、2g(E2)、3g(E3)、4g(E4)、5g(E5)、6g(E6) and 7g (E7) group, 9 processing group difference
The feed for feeding the mulberry leaf ether extract Han 0,0,1,2,3,4,5,6 and 7g/kg is fed 60 days.After 60 days measure crucian carp growth,
Food ration and feed efficiency and weight gain recovery rate broken line regression analysis is carried out, the result is shown in table 3 and Fig. 8.Data are in table 3 with 4
Duplicate average ± standard deviation indicates；In same index, the female difference person of data shoulder marking-up indicates significant difference between data, phase
Indicate that difference is not significant (P < 0.05) with person.Data are indicated in Fig. 8 with 4 duplicate averages.Weight gain recovery rate (%)=
(E-Y)/(D-Y) × 100 (table 3).
Table 3 feeds influence of the feed of ether extract containing mulberry leaf (EEE) to crucian high density stress
As shown in Table 3, the feed for feeding the ether extract containing mulberry leaf has restored growth, food ration and the feed efficiency of fish.
As shown in Figure 8, the best addition concentration of the anti-cultivation high density stress of mulberry leaf ether extract is 4.85g/kg feed.
Embodiment 9
With under the identical breeding environment of embodiment 5,21.2 ± 1.1g of weight carp fry, 210 tail is selected, is randomly divided into 7
Processing group, every group of 3 fish jars, every 10 tail fish of cylinder.Copper sulphate is dissolved in 7 processing group fish tank water bodies, makes its concentration be respectively
0.0,0.4,0.5,0.6,0.7,0.8 and 0.9mg/L, 7 processing groups feed fish meal bean pulp type omnivorefish feed, after four days
The feed intake and the death rate of each processing group are investigated, experimental result is shown in Fig. 9 and 10.Wherein, data are duplicate average with 3 in figure
Number ± standard deviation indicates；Reference character difference person indicates significant difference between data on column, identical person indicate difference it is not significant (P <
0.05)。
By Fig. 9 and 10 it is found that as the food ration of the raising fish of Cu concentration gradually decreases；When Cu concentration increases to 0.7mg/L
When water, fish food ration drops to floor level；When further increasing Cu concentration up to 0.8mg/L, fish occurs dead.Meanwhile it generating
Maximum copper sulphate stress be without causing dead Cu concentration to be 0.7mg/L.
Embodiment 10
With under the identical breeding environment of embodiment 5,210 tail of same body weight carp fry is selected, 7 processing are randomly divided into
Group, every group of 3 fish jars, every 10 tail fish of cylinder.Metrifonate is dissolved respectively in 7 processing group fish tank water bodies, reaches its concentration
0.0,0.4,1.0,1.6,2.2,2.8 and 3.4mg/L.The fish rollover rate and the death rate of each processing group are investigated after four days, are tied
Fruit sees Figure 11 and 12.Wherein, data are indicated in figure with 3 duplicate average ± standard deviations；Reference character difference person table on column
Registration significant difference between, identical person indicate that difference is not significant (P < 0.05).
By Figure 11 and 12 it is found that as the rollover rate of the raising fish of metrifonate concentration gradually rises, when metrifonate concentration increases
When to 2.2mg/L, fish rollover rate reaches highest level, and when further increasing metrifonate concentration up to 2.8mg/L, fish occurs dead.
Cause maximum metrifonate stress be without causing the concentration of fish death to be 2.2mg/L water.
Embodiment 11
With under the identical breeding environment of embodiment 5,10.5 ± 0.3g of weight carp fry, 480 tail is selected, is randomly divided into 8
Processing group, every group of 4 fish jars, every 15 tail fish of cylinder.8 processing groups feed 0,1,2,3,4,5,6 and 7g/kg mulberry leaf ether respectively and mention
Take object feed.After 30 days, 30 tail fishes are randomly choosed from each processing group respectively, averagely give 3 fish jars (every 10 tail fish of cylinder)；?
Copper sulphate is added in each fish jar water body makes its concentration 0.7mg Cu/L.Feed fish meal bean pulp type omnivorefish feed, four days
The grazing rate of each processing group is investigated afterwards and carries out recovery rate broken line regression analysis of ingesting, and the result is shown in table 4 and Figure 13.The number of table 4
3 duplicate average ± standard deviations indicate accordingly；Superscript letters difference person indicates significant difference in same column data, identical
Person indicates that difference is not significant (P < 0.05).Data are indicated in Figure 13 with 3 duplicate averages.Grazing rate (%)=food ration/
Weight × 100, recovery rate of ingesting (%)=(E-Y)/(K-Y) × 100.
Table 4 feed mulberry leaf ether extract (EEE) to copper sulphate stress rear crucian grazing rate influence
As shown in Table 4, feed the ether extract containing mulberry leaf feed restored sulfuric acid copper inducible crucian grazing rate.By
Figure 13 is it is found that the best addition concentration that mulberry leaf ether extract restores crucian grazing rate is 4.52g/kg feed.
Embodiment 12
With under the identical breeding environment of embodiment 5,10.5 ± 0.3g of weight carp fry, 480 tail is selected, is randomly divided into 8
Processing group, every group of 4 fish jars, every 15 tail fish of cylinder.8 processing groups feed 0,1,2,3,4,5,6 and 7g/kg mulberry leaf ether respectively and mention
Take object feed.After 30 days, 30 tail fishes are randomly choosed from each processing group respectively, averagely give 3 fish jars (every 10 tail fish of cylinder),
In each fish jar water body plus metrifonate makes its concentration 2.2mg/L.Investigated after four days each processing group rollover fish quantity and into
Row rollover inhibiting rate broken line regression analysis, the result is shown in table 5 and Figure 14.Data are in table 5 with 3 duplicate average ± standard deviations
It indicates, each repetition is the average value of 4 days rollover rates；Superscript letters difference person indicates significant difference, phase in same column data
Indicate that difference is not significant (P < 0.05) with person.Data are indicated in Figure 14 with 3 duplicate averages.Rollover rate (%)=rollover fish
Quantity/fish total quantity × 100.
Table 5 feed mulberry leaf ether extract (EEE) to metrifonate stress rear crucian rollover rate influence
As shown in Table 5, the crucian rollover that mulberry leaf ether extract inhibits metrifonate to induce is fed.Mulberry leaf as shown in Figure 14
It is 4.90g/kg feed that ether extract, which inhibits the best addition concentration of crucian rollover,.
Although being described in detail in conjunction with attached drawing to a specific embodiment of the invention, should not be construed as special to this
The restriction of the protection scope of benefit.In range described by claims, those skilled in the art are without creative work
The various modifications and deformation that can make still belong to the protection scope of this patent.
Claims (10)
1. mulberry-leaf extract inhibit freshwater fish culturing stress aspect application.
2. mulberry-leaf extract as described in claim 1 inhibit freshwater fish culturing stress aspect application, which is characterized in that will
Mulberry-leaf extract, which is added in feed, to be used, and adding proportion is 4.52~7.00wt ‰.
3. mulberry-leaf extract as claimed in claim 1 or 2 inhibit freshwater fish culturing stress aspect application, feature exists
In the mulberry-leaf extract is mulberry leaf ether extract, mulberry leaf hexamethylene extract or mulberry leaf acetone extract.
4. mulberry-leaf extract as claimed in claim 3 inhibit freshwater fish culturing stress aspect application, which is characterized in that institute
It states mulberry leaf hexamethylene extract to be made by the following method: by mulberry leaf drying, crush, Mulberry Leaf and hexamethylene are compared by 1:5~8
Example mixes, and stirring is filtered after extracting, and repeats to extract 2~3 times, obtains filtrate and the first filter residue, is evaporated under reduced pressure after merging filtrate to perseverance
Weight is made；
The mulberry leaf ether extract is made by the following method: will be first obtained in mulberry leaf hexamethylene extract extraction process
Filter residue and ether are mixed in the ratio of 1:5~8, and stirring is filtered after extracting, and repeat to extract 2~3 times, obtain no residue filtrate and the second filter
Slag, vacuum distillation is made to constant weight after merging filtrate；
The mulberry leaf acetone extract is made by the following method: by the second filter obtained in mulberry leaf ether extract extraction process
Slag and acetone are mixed in the ratio of 1:5~8, and stirring is filtered after extracting, and repeat to extract 2~3 times, obtain no residue filtrate and third filter
Slag, vacuum distillation is made to constant weight after merging filtrate.
5. a kind of fresh-water fishes Antistress agent, which is characterized in that including mulberry-leaf extract described in claim 3 or 4.
6. a kind of feed, which is characterized in that including mulberry-leaf extract described in basal feed and claim 3 or 4.
7. feed as claimed in claim 6, which is characterized in that including following parts by weight of component: it is total that mulberry-leaf extract accounts for feed
The 4.52 of weight~7.00 ‰.
8. feed as claimed in claim 7, which is characterized in that including following parts by weight of component: it is total that mulberry-leaf extract accounts for feed
The 4.90 ‰ of weight.
9. such as the described in any item feeds of claim 6~8, which is characterized in that the basal feed includes following parts by weight group
Point: 23.00~25.00 parts of fish meal, 31.00~33.00 parts of dregs of beans, 0.60~0.80 part of DL- METHIONINE, flour 36.00~
38.00 parts, 3.00~3.50 parts of grease, 0.80~1.30 part of vitamine supplement and 0.80~1.30 part of mineral additive.
10. such as the preparation method of the described in any item feeds of claim 6~9, which comprises the following steps:
Mulberry-leaf extract is dissolved in grease, is then mixed with remaining component of basal feed, is pelletized, is made.
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| CN111084301A (en) * | 2019-12-30 | 2020-05-01 | 内江师范学院 | Fish feed adhesive containing mulberry leaf extract as well as preparation method and application of fish feed adhesive |
| CN111772057A (en) * | 2020-07-23 | 2020-10-16 | 内江师范学院 | Natural compound for improving fish anti-stress response and application thereof |
| CN111802535A (en) * | 2020-07-23 | 2020-10-23 | 内江师范学院 | Natural composition for improving fish anti-stress response and application thereof |
| KR102258211B1 (en) * | 2020-12-04 | 2021-05-31 | 어업회사법인 부안수산 | Aquatic product farming method using food including mulberry leaves |
| CN115316555A (en) * | 2022-07-01 | 2022-11-11 | 内江师范学院 | Feed for eliminating heavy metal cadmium in crucian body |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111084301A (en) * | 2019-12-30 | 2020-05-01 | 内江师范学院 | Fish feed adhesive containing mulberry leaf extract as well as preparation method and application of fish feed adhesive |
| CN111084301B (en) * | 2019-12-30 | 2023-08-22 | 内江师范学院 | Mulberry leaf extract fish feed adhesive and preparation method and application thereof |
| CN111772057A (en) * | 2020-07-23 | 2020-10-16 | 内江师范学院 | Natural compound for improving fish anti-stress response and application thereof |
| CN111802535A (en) * | 2020-07-23 | 2020-10-23 | 内江师范学院 | Natural composition for improving fish anti-stress response and application thereof |
| CN111772057B (en) * | 2020-07-23 | 2023-05-09 | 内江师范学院 | Natural compound for improving anti-stress response of fish and application thereof |
| CN111802535B (en) * | 2020-07-23 | 2023-05-09 | 内江师范学院 | Natural composition for improving anti-stress response of fish and application thereof |
| KR102258211B1 (en) * | 2020-12-04 | 2021-05-31 | 어업회사법인 부안수산 | Aquatic product farming method using food including mulberry leaves |
| CN115316555A (en) * | 2022-07-01 | 2022-11-11 | 内江师范学院 | Feed for eliminating heavy metal cadmium in crucian body |
| CN115316555B (en) * | 2022-07-01 | 2023-08-18 | 内江师范学院 | Feed for eliminating heavy metal cadmium in crucian bodies |
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