CN115067439A - Grass carp immunity performance feed based on reed and preparation method thereof - Google Patents
Grass carp immunity performance feed based on reed and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a grass carp immunity performance feed based on bulrush and a preparation method thereof, wherein the feed comprises the following raw materials in percentage by mass: 0-30% of wheat, 0-30% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of calcium dihydrogen phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil; the feed mainly comprises wheat, reed, bean pulp, fish meal, rapeseed meal, monocalcium phosphate, CMC (carboxy methyl cellulose) adhesive, choline chloride, bentonite, yttrium oxide, vitamin premix, mineral salt premix and soybean oil, and the active ingredients in the reed can improve the nonspecific immunoreaction level of grass carp to a certain extent by adding the reed into the feed.
Description
Technical Field
The invention belongs to the technical field of fish feed production, and particularly relates to a grass carp immunity feed based on reeds and a preparation method thereof.
Background
Fish as a lower vertebrate has perfect acquired immune response, and can resist pathogen invasion through nonspecific immune response and specific immune response. The feed is an important component of a fish nonspecific immune system, is mainly synthesized by liver, has the functions of bacteriolysis and cell lysis, and plays an important role in the early anti-infection process. IL-1 is an important cell factor of fishes, and plays an important role in regulating phagocyte activity, inducing inflammatory response and resisting pathogen invasion. Lysozyme is an important nonspecific immune factor in fish bodies and plays an important role in removing and killing pathogenic microorganisms. The IFN system is the earliest immune factor of a cell function regulating system in the body defense response at present; IgM is a specific immune factor produced by B lymphocyte differentiation that is involved in the inhibition of pathogenic activity and regulation of antigen presentation processes.
However, the existing fish feed usually contains wheat ingredients, the wheat affects the liver structure and the immunity of the grass carp, and the wheat has an undesirable effect on the liver amino acid metabolism capability of the grass carp, so that the nonspecific immune response level effect of the grass carp is undesirable, and therefore, the grass carp immunity feed based on the reed and the preparation method thereof are needed to solve the existing problems.
Disclosure of Invention
The invention aims to provide a grass carp immunity performance feed based on reeds and a preparation method thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a grass carp immunity performance feed based on reed comprises the following raw materials by mass percent: 0-30% of wheat, 0-30% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of monocalcium phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil.
Preferably, the material comprises the following raw materials in percentage by mass: 30% of wheat, 0% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of monocalcium phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil.
Preferably, the material comprises the following raw materials in percentage by mass: 15% of wheat, 15% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of monocalcium phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil.
Preferably, the material comprises the following raw materials in percentage by mass: 0% of wheat, 30% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of monocalcium phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil.
Preferably, when the reed is used, the reed is ground into powder after being crushed and dried at 56 ℃, the effective components of the reed comprise water, crude protein, crude ash powder and crude fiber, and the total energy of the reed is 17.73 MJ/kg.
Preferably, the vitamin premix comprises the following raw materials in parts by mass: 1.65 parts of vitamin A, 0.025 part of vitamin D, 50 parts of vitamin E and 10 parts of vitamin K100 parts of vitamin C, 20 parts of thiamine, 20 parts of riboflavin and vitamin B 6 20 portions of vitamin B 12 0.02 part, 5 parts of folic acid, 50 parts of calcium pantothenate, 100 parts of nicotinic acid, 0.1 part of biotin and 645.2 parts of cellulose.
Preferably, the mineral salt premix comprises the following raw materials in parts by mass: 500 parts of sodium chloride, 8155.6 parts of magnesium sulfate heptahydrate, 12500 parts of sodium dihydrogen phosphate dihydrate, 16000 parts of potassium dihydrogen phosphate, 7650.6 parts of calcium hydrophosphate dihydrate, 2286.2 parts of ferrous sulfate heptahydrate, 1750 parts of calcium lactate pentahydrate, 178 parts of zinc sulfate heptahydrate, 61.4 parts of manganese sulfate monohydrate, 15.5 parts of copper sulfate pentahydrate, 0.5 part of cobalt sulfate heptahydrate, 1.5 parts of potassium iodide and 753.7 parts of corn starch.
Based on the grass carp immunity performance feed based on the reed, the invention also provides a preparation method of the grass carp immunity performance feed based on the reed, which comprises the following steps:
s1, preparing the following raw materials: wheat, reed, soybean meal, fish meal, rapeseed meal, monocalcium phosphate, CMC (carboxy methyl cellulose) adhesive, choline chloride, bentonite, yttrium oxide, vitamin premix, mineral salt premix and soybean oil;
s2, respectively crushing the solid raw materials of the wheat, the reed, the bean pulp and the rapeseed pulp, sieving the crushed mixture, and selecting solid particles with the diameter less than 0.43 mm;
s3, mixing and stirring the solid particles according to the feed ratio to obtain a feed mixture;
s4, processing the feed mixture into feed particles with the diameter of 3mm by using a feed granulator;
and S5, packaging the feed particles, and storing the feed particles in a cool and dry environment for later use.
Preferably, the nutrient components of the feed particles in the step S4 comprise 33% -32.61% of crude protein, 4.6% -4.82% of crude fat, 4.84% -13.87% of crude fiber and 9.95% -12.45% of ash, and the total nutrient content of the feed particles is 19.84% -19.96%.
Compared with the prior art, the grass carp immunity performance feed based on the reed and the preparation method thereof provided by the invention have the following advantages:
the feed mainly comprises wheat, reed, bean pulp, fish meal, rapeseed meal, monocalcium phosphate, CMC (carboxy methyl cellulose) adhesive, choline chloride, bentonite, yttrium oxide, vitamin premix, mineral salt premix and soybean oil, and the active ingredients in the reed can improve the nonspecific immunoreaction level of grass carp to a certain extent by adding the reed into the feed.
Drawings
FIG. 1 is a block flow diagram of the present invention;
FIG. 2 is a schematic diagram showing the effect of Phragmites communis on the content and activity of immune factors in serum and liver of grass carp;
FIG. 3 is a schematic diagram showing the effect of Phragmites communis on the expression level of immune genes in head and kidney (A), liver (B), spleen (C) and kidney (D) tissues of grass carp;
FIG. 4 is a schematic diagram showing the effect of Phragmites communis on the liver tissue morphology of grass carp.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. The specific embodiments described herein are merely illustrative of the invention and do not delimit the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The invention provides a grass carp immunity performance feed based on bulrush, which comprises the following raw materials in percentage by mass: 30% of wheat, 0% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of monocalcium phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil;
when the reed is used, the reed is crushed, dried at 56 ℃ and ground into powder, the effective components of the reed comprise water, crude protein, crude ash powder and crude fiber, and the total energy of the reed is 17.73 MJ/kg.
The vitamin premix comprises the following raw materials in parts by mass: 1.65 parts of vitamin A, 0.025 part of vitamin D, 50 parts of vitamin E, 10 parts of vitamin K, 100 parts of vitamin C, 20 parts of thiamine, 20 parts of riboflavin and vitamin B 6 20 portions of vitamin B 12 0.02 part, 5 parts of folic acid, 50 parts of calcium pantothenate, 100 parts of nicotinic acid, 0.1 part of biotin and 645.2 parts of cellulose.
The mineral salt premix comprises the following raw materials in parts by mass: 500 parts of sodium chloride, 8155.6 parts of magnesium sulfate heptahydrate, 12500 parts of sodium dihydrogen phosphate dihydrate, 16000 parts of potassium dihydrogen phosphate, 7650.6 parts of calcium hydrophosphate dihydrate, 2286.2 parts of ferrous sulfate heptahydrate, 1750 parts of calcium lactate pentahydrate, 178 parts of zinc sulfate heptahydrate, 61.4 parts of manganese sulfate monohydrate, 15.5 parts of copper sulfate pentahydrate, 0.5 part of cobalt sulfate heptahydrate, 1.5 parts of potassium iodide and 753.7 parts of corn starch.
Based on the grass carp immunity performance feed based on the reed, the invention also provides a preparation method of the grass carp immunity performance feed based on the reed, which comprises the following steps:
s1, preparing the following raw materials: wheat, reed, soybean meal, fish meal, rapeseed meal, monocalcium phosphate, CMC (carboxy methyl cellulose) adhesive, choline chloride, bentonite, yttrium oxide, vitamin premix, mineral salt premix and soybean oil;
s2, respectively crushing the solid raw materials of the wheat, the reed, the bean pulp and the rapeseed pulp, sieving the crushed mixture, and selecting solid particles with the diameter less than 0.43 mm;
s3, mixing and stirring the solid particles according to the feed ratio to obtain a feed mixture;
s4, processing the feed mixture into feed particles with the diameter of 3mm by using a feed granulator; the nutrient components of the feed particles comprise 33 to 32.61 percent of crude protein, 4.6 to 4.82 percent of crude fat, 4.84 to 13.87 percent of crude fiber and 9.95 to 12.45 percent of ash, and the total nutrient content of the feed particles is 19.84 to 19.96 percent;
and S5, packaging the feed particles, and storing the feed particles in a cool and dry environment for later use.
Example 2
The same parts are not repeated, and different from the embodiment 1, the coating comprises the following raw materials by mass percent: 15% of wheat, 15% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of monocalcium phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil.
Example 3
The same parts are not repeated, and different from the embodiment 1, the coating comprises the following raw materials by mass percent: 0% of wheat, 30% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of monocalcium phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil.
Example 4
The feed ingredient ratios according to example 1, example 2 and example 3 were as follows: (namely, compared with wheat, the content of the reed in the embodiment 1 is 0%, the content of the reed in the embodiment 2 is 50%, and the content of the reed in the embodiment 3 is 100%)
| Categories | Example 1 | Example 2 | Example 3 |
| Wheat (Triticum aestivum L.) | 30％ | 15％ | 0％ |
| |
0％ | 15％ | 30％ |
| Bean pulp | 30％ | 30％ | 30％ |
| |
3％ | 3％ | 3％ |
| Rapeseed dregs | 26％ | 26％ | 26％ |
| Calcium dihydrogen phosphate | 2.5％ | 2.5％ | 2.5％ |
| |
2％ | 2％ | 2％ |
| Choline chloride | 0.15％ | 0.15％ | 0.15％ |
| Bentonite clay | 1.25％ | 1.25％ | 1.25％ |
| Yttrium oxide | 0.1％ | 0.1％ | 0.1％ |
| |
1％ | 1％ | 1％ |
| |
1％ | 1％ | 1％ |
| Soybean |
3％ | 3％ | 3％ |
Selecting 66 healthy grass carps, randomly dividing the grass carps into 3 groups, feeding the feeds prepared in the embodiment 1, the embodiment 2 and the embodiment 3 for 3 times, and after feeding the grass carps for 41 days, respectively detecting the changes of serum biochemical indexes, liver tissue structures and immune related indexes, wherein the specific detection process and results are as follows:
1.2 Breeding management
The cultivation test was carried out in an indoor recirculating aquaculture plastic drum (diameter 1.5 m). The grass carp used for the test is bred in a breeding farm and belongs to large-size grass carps, after the grass carps are temporarily bred for 2 weeks by using basic feed, 66 healthy grass carps with the average weight of (145 +/-2) g are selected and randomly divided into 3 groups (P is more than 0.05) according to the difference of body mass, 3 repeats are set in each group, and 7-8 repeats are set in each group. Feeding is carried out 2 times in the morning (8-9 o 'clock) and afternoon (3-4 o' clock) every day in a breeding test, the feeding amount in the initial day is 3% of the weight of the fish, and the breeding period is 41 d. And (3) maintaining micro flowing water during the culture period, aerating by an air compressor, measuring the water temperature every day, and observing the activity condition, the health state and the like of the experimental fish.
1.3 sample Collection and processing
Stopping feeding for 24h after the culture experiment is finished, randomly selecting 3 fishes in each water tank, anticoagulating by using heparin sodium, taking blood from tail veins, standing for 1h at 4 ℃, centrifuging for 15min at 4000r/min, sucking supernatant, and taking supernatant for biochemical index and immune factor content detection; after dissection, taking about 0.2g of liver, spleen, kidney and head and kidney respectively, quickly freezing by liquid nitrogen, and storing in a refrigerator at-80 ℃ for immune gene expression level detection; taking about 1g of liver, putting the liver into a 2mL centrifuge tube, adding 1mL of sterilized ddH2O to prepare liver tissue homogenate, diluting by 10 times, subpackaging 150 mu L of each tube, and storing in a refrigerator at-80 ℃ for detecting the content of immune factors in the liver; liver tissues of 0.5cm × 0.5cm × 0.5cm size were fixed in 10% formaldehyde for H & E staining and liver tissue structure was observed.
1.4 serum Biochemical index detection
The detection of the biochemical indexes of the serum is finished by a Heidel commercial source medical detection center, Total Protein (TP) is measured by a biuret method, Albumin (ALB) is measured by a BCG method, Globulin (GLB) is measured by a calculation method, blood sugar (GLU) is measured by a hexokinase method, and the activities of glutamic-oxalacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT) are measured by an IFCC rate method.
1.5 Observation of liver tissue Structure
Fixing liver tissue in 10% formaldehyde for 24h, performing gradient dehydration with 30% -100% ethanol, xylene transparence and wax immersion embedding to prepare paraffin tissue slices with the thickness of 4-5 um, performing HE conventional dyeing, sealing with neutral gum, and observing structural characteristics of the liver tissue by using a Leica advanced microsystem (DM 3000).
1.6 detection of immune factor content and Activity
The content detection of immunoglobulin M (IgM), interleukin 1(IL-1), complement protein 3(C3), total complement (CH50) and Cortisol (Cortisol) in serum and liver is carried out by adopting an ELISA kit of corresponding fish immune factors and a detection method reference kit.
1.7 assay of immune Gene expression level
1.7.1RNA extraction and first Strand cDNA Synthesis
Liver, spleen, kidney and head and kidney tissues preserved at-80 ℃ were taken, total RNA was extracted using TRIzol reagent (Invitrogen, USA) according to the method described in reference to TRIzol reagent specification, RNA integrity was detected using 1% agarose electrophoresis of the extracted RNA, and total RNA with good integrity was used in the next experiment. 2 μ g of total RNA was taken and RevertAid was used TM First StrandcDNASynthesis (ThermoFisher, USA) reverse transcription kit, using oligo (dT)18 primer to synthesize cDNA first strand for analysis of immune gene expression level.
1.7.2 fluorescent quantitative PCR analysis
Referring to relevant literature, the immune genes detected in the experiment are determined to be heat stress protein 70(Hsp70), lysozyme, major histocompatibility complex I (MHC-I), Interferon (IFN), C3, IgM and IL-1. The primer synthesis is completed by the company of Biotechnology engineering (Shanghai), Inc., and the sequence information of the primers is shown in the table below:
fluorescent quantitative PCR reaction system: 2 XMastermix 10. mu.L, 10. mu. mol/L primer 0.4. mu.L, diluted template 9.6. mu.L. The reaction procedure is as follows: pre-denaturation at 95 ℃ for 3 min: 5s at 95 ℃, 10s at 60 ℃ and 15s at 72 ℃ for 40 cycles, the dissolution curve is 65-97 ℃, and the temperature rises by 0.1 ℃ per second. The gene expression level is calculated by adopting a 2-delta Ct method, light cycler480software elease1.5.0 and Excel software are used for data processing, an actin gene is used as a reference gene, and the expression level of 0% of the substitution group genes is subjected to homogenization analysis.
1.8 data analysis
SPSS17.0 software is adopted to carry out data One-way ANOVA, Duncan method is adopted to carry out significance analysis among groups, the significance level is P <0.05, the detection result is the mean value plus or minus standard deviation (mean plus or minus SE), and the detection result of the content and the activity of the immune factor and the expression level of the immune related gene is plotted by GraphPadprism6 software.
2. Results and analysis
2.1 serum Biochemical indicators
The content of TP, ALB and GLB in the serum of the grass carp shows a trend of increasing firstly and then decreasing with the increase of the substitution proportion of the reed, but the content of TP, ALB and GLB among 3 groups has no obvious change (P is more than 0.05) on the whole. The blood sugar concentration of the grass carps in the substitution group is lower than that of the control group, and particularly, the blood sugar concentration of 50% of the substitution group is obviously lower than that of the control group. Besides the AST activity of 100% of the substitution group, the ALT and AST activities in the serum of the grass carp in the substitution group are greatly increased and are remarkably higher than those in a control group (P <0.05), and the following table shows the influence of reed on biochemical indexes of the serum of the grass carp:
| |
0％ | 50％ | 100％ |
| Total protein TP (g/L) | 32.97±3.29 | 36.50±1.87 | 34.57±0.35 |
| Albumin ALB (g/L) | 15.73±1.11 | 17.00±1.04 | 15.70±0.40 |
| Globulin GLB/(g/L) | 17.23±2.18 | 19.50±0.90 | 18.87±0.64 |
| Blood sugar GLU (mmol/L) | 9.85±0.81a | 6.35±0.56b | 7.68±1.02ab |
| Alanine aminotransferase ALT/(U/L) | 22.35±2.05a | 64.60±8.34b | 98.05±1.48c |
| Glutamic-oxaloacetic transaminase AST/(U/L) | 47.00±3.11a | 84.10±17.25b | 51.45±0.92a |
Note: the difference of the superscript letters of the same row data indicates that the difference between two groups is significant (P <0.05), and the same indicates that the difference between two groups is not significant (P > 0.05).
2.2 liver tissue Structure
The control group had poorly defined liver cells and severe vacuolization of liver cells, which was accompanied by a small amount of lymphocyte infiltration (a, B in fig. 4). The liver cells of the grass carp livers of the 50% and 100% substitution groups are relatively clear, the cell nuclei are clearly developed and basically located in the center, and the cells are regularly arranged and uniform in size (C, D, E and F in the figure 4).
2.3 immune factor content and Activity
Compared with the control group, the content of IL-1 and C3 is obviously increased in grass carp serum of the 100% reed replacement group (P <0.05), and the content of CH50 is obviously lower than that of the control group (P <0.05) in both 50% and 100% reed replacement groups. The content of IL-1 and CH50 in the liver of the grass carp in the 100% reed replacement group is obviously higher than that in the control group (P <0.05), but the content of C3 is obviously lower than that in the control group (P < 0.05); the content changes of IL-1, C3, IgM and Cortisol in the serum and liver of the grass carp in the 50% reed replacement group are not significant (P is more than 0.05), the influence of reed on the content and activity of immune factors in the serum and liver of the grass carp is shown in figure 2, and the units of the content and the activity of the immune factors are IgM (mu g/mL), IL-1(pg/mL), C3 (mu g/mL), CH50(U/mL) and Cortisol (ng/mL) respectively.
2.4 immune Gene expression levels
Compared with a control group, Hsp70, IL-1 and MHC-I, Lysozyme in the 100% reed-substituted group are all remarkably up-regulated in head and kidney (P <0.05), and C3 and IFN are both remarkably down-regulated; the 50% reed replacing grass carp C3, Hsp70, Lysozyme, MHC-I, IFN and other genes are obviously down-regulated in head and kidney (P <0.05) (figure 3-A); IL-1 and Lysozyme in the liver of grass carp in the substitute group were all significantly reduced in expression compared with the control group (P <0.05), while the differences in expression levels of C3, MHC-I, IFN and IgM were all insignificant (P >0.05) (FIG. 3-B); the expression level of C3 and Hsp70 in the spleen of grass carp in the 50% reed replacement group is significantly higher than that of a control group, and the expression level of MHC-I is significantly lower than that of the control group (P < 0.05); except IFN and IgM genes in the spleen of the grass carp in the 100% reed replacement group, the expression level of other detection genes is obviously lower than that of a control group (figure 3-C); the mRNA expression levels of Hsp70, MHC-1, IFN and IgM genes in the kidney of the grass carp in the 100% reed replacement group are all significantly higher than those of the control group (P <0.05), and the other detected gene expression levels except C3 in the kidney of the 50% reed replacement group are not significantly different (P >0.05) (FIG. 3-D).
3. Influence of reeds with different substitution ratios on immunity of grass carp
IL-1, C3 and Cortisol in serum of grass carp in the substitution group, and IL-1 and CH50 in liver show a trend rising along with the increase of the substitution proportion of reed, wherein the change of the nonspecific immunity index of the grass carp in the 100% reed substitution group is significantly different from that of the control group (P <0.05), and the IgM content is not significantly different (P > 0.05). According to the results of the immune factor content change, a proper amount of reed is added into the feed, so that the nonspecific immune reaction of the grass carp can be enhanced, the influence on the specific immune reaction is not obvious, the results are consistent with the research results of [30] such as aging and the like, and the active ingredient rutin in the reed can be found to remarkably improve the nonspecific immunity of the grass carp. Meanwhile, the result of the change of the immune gene expression level shows that the feed added with 100 percent of reed can up-regulate the expression levels of Hsp70, IL-1, Lysozyme, MHC-I, IFN and IgM in head and kidney to different degrees, and the 50 percent of reed replacing group grass carp immune gene expression level is down-regulated compared with a control group. Therefore, the result of the trend of the mRNA expression level change of the grass carp immunity gene shows that the grass carp immunity level tends to decrease and then increase along with the increase of the addition amount of the reed, and a more detailed experimental method needs to be designed for exploring the regulation mechanism of the influence of the reed on the grass carp immunity. Therefore, the results of the comprehensive immune factor content and mRNA expression level change show that the addition of reed in the feed can improve the nonspecific immune response level of grass carp to a certain extent, which may be related to flavonoid, reed polysaccharide, rutin and the like which are active ingredients contained in reed. Researches show that the flavonoid active substances in the reed have remarkable antibacterial capacity and antioxidant performance; the tender reed contains abundant vitamins, plays an important role in organism metabolism, and can enhance the immunity of the rex rabbit.
In conclusion, the results show that the glutamic-pyruvic transaminase and the glutamic-oxalacetic transaminase in the serum of the grass carp in the replacement group are obviously increased compared with the control group, and the total protein, the albumin and the globulin are not obviously changed compared with the control group. The liver tissue section result shows that compared with the control group, the grass carp liver cell boundary of the substitute group is clear, and the tissue structure is improved. The detection results of the immune factor content and the expression level show that compared with the control group, the serum content of interleukin 1(IL-1) and complement 3(C3) in the grass carp of the 100% substitution group is obviously increased (P <0.05), and the content of IL-1 and total complement (50% Haemolyticcomplete, CH50) in the liver is obviously increased (P < 0.05); the content of immune factors such as IL-1 and C3 in serum and IL-1 and CH50 in liver of 50 percent reed replacing grass carp is higher than that of a control group; meanwhile, the relative expression levels of C3, IL-1, major histocompatibility complex I (MHC-I), Interferon (IFN) and other genes in head, kidney, liver and spleen of the grass carp can be obviously influenced by feeding feeds with different reed replacing ratios. Therefore, the reed instead of wheat can improve the liver tissue structure of the grass carp, improve the metabolic capability of liver amino acid and improve the nonspecific immunoreaction level of the grass carp to a certain extent.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments or portions thereof without departing from the spirit and scope of the invention.
Claims (9)
1. A grass carp immunity performance feed based on reed is characterized in that: the composite material comprises the following raw materials in percentage by mass: 0-30% of wheat, 0-30% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of calcium dihydrogen phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil.
2. The grass carp immunity performance feed based on reed as claimed in claim 1, wherein: the composite material comprises the following raw materials in percentage by mass: 30% of wheat, 0% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of monocalcium phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil.
3. The grass carp immunity performance feed based on reed as claimed in claim 1, wherein: the composite material comprises the following raw materials in percentage by mass: 15% of wheat, 15% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of calcium dihydrogen phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil.
4. The grass carp immunity performance feed based on bulrush as claimed in claim 1, wherein: the composite material comprises the following raw materials in percentage by mass: 0% of wheat, 30% of reed, 30% of soybean meal, 3% of fish meal, 26% of rapeseed meal, 2.5% of monocalcium phosphate, 2% of CMC adhesive, 0.15% of choline chloride, 1.25% of bentonite, 0.1% of yttrium oxide, 1% of vitamin premix, 1% of mineral salt premix and 3% of soybean oil.
5. The grass carp immunity performance feed based on reed as claimed in claim 1, wherein: when the reed is used, the reed is crushed, dried at 56 ℃ and ground into powder, the effective components of the reed comprise water, crude protein, crude ash powder and crude fiber, and the total energy of the reed is 17.73 MJ/kg.
6. The reed-based grass carp immunity performance feed as claimed in claim 5, wherein: the vitamin premix comprises the following raw materials in parts by mass: 1.65 parts of vitamin A, 0.025 part of vitamin D, 50 parts of vitamin E, 10 parts of vitamin K, 100 parts of vitamin C, 20 parts of thiamine, 20 parts of riboflavin, and vitamin B 6 20 portions of vitamin B 12 0.02 part, 5 parts of folic acid, 50 parts of calcium pantothenate, 100 parts of nicotinic acid, 0.1 part of biotin and 645.2 parts of cellulose.
7. The grass carp immunity performance feed based on reed as claimed in claim 1, wherein: the mineral salt premix comprises the following raw materials in parts by mass: 500 parts of sodium chloride, 8155.6 parts of magnesium sulfate heptahydrate, 12500 parts of sodium dihydrogen phosphate dihydrate, 16000 parts of potassium dihydrogen phosphate, 7650.6 parts of calcium hydrogen phosphate dihydrate, 2286.2 parts of ferrous sulfate heptahydrate, 1750 parts of calcium lactate pentahydrate, 178 parts of zinc sulfate heptahydrate, 61.4 parts of manganese sulfate monohydrate, 15.5 parts of copper sulfate pentahydrate, 0.5 part of cobalt sulfate heptahydrate, 1.5 parts of potassium iodide and 753.7 parts of corn starch.
8. A method for preparing a reed-based grass carp immunity performance feed according to any one of claims 1-7, which is characterized in that: the method comprises the following steps:
s1, preparing the following raw materials: wheat, reed, soybean meal, fish meal, rapeseed meal, monocalcium phosphate, CMC (carboxy methyl cellulose) adhesive, choline chloride, bentonite, yttrium oxide, vitamin premix, mineral salt premix and soybean oil;
s2, respectively crushing the solid raw materials of the wheat, the reed, the bean pulp and the rapeseed pulp, sieving the crushed mixture, and selecting solid particles with the diameter less than 0.43 mm;
s3, mixing and stirring the solid particles according to the feed ratio to obtain a feed mixture;
s4, processing the feed mixture into feed particles with the diameter of 3mm by using a feed granulator;
and S5, packaging the feed particles, and storing the feed particles in a cool and dry environment for later use.
9. The preparation method of the grass carp immunity performance feed based on the reed as claimed in claim 8, wherein the preparation method comprises the following steps: the nutrient components of the feed particles in the step S4 comprise 33-32.61% of crude protein, 4.6-4.82% of crude fat, 4.84-13.87% of crude fiber and 9.95-12.45% of ash, and the total nutrient content of the feed particles is 19.84-19.96%.
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