CN112741221A - Immunity-improving cat food and preparation method thereof - Google Patents
Immunity-improving cat food and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a cat food for improving immunity and a preparation method thereof. Sodium alginate is modified by adopting Maillard reaction to prepare the sodium alginate-glycine copolymer, so that the capacity of removing free radicals of the sodium alginate is improved. The liposome of the crab methallyl alcohol glycoside is modified by modified montmorillonite and then prepared into microspheres with sodium alginate-glycine copolymer, so that the biological activity of the crab methallyl alcohol glycoside can be protected. The sodium alginate has the effects of reducing sugar and fat, and can be used for protecting liver of cat in cooperation with crab methallyl alcohol glycoside, and reducing burden of high fat diet on liver of cat; the gene of the cat interferon is recombined into the lactobacillus plantarum body by utilizing a genetic engineering means to obtain the lactobacillus plantarum producing the cat interferon, the lactobacillus plantarum is used for fermenting fish meat to obtain fish meat containing the interferon, and the immunity of a cat is improved after the fish meat is eaten.
Description
Technical Field
The invention relates to the technical field of foods, and particularly relates to a cat food for improving immunity and a preparation method thereof.
Background
Kittens are complete carnivores, so their primary energy and nutrient source should come from animal proteins and fats, rather than carbohydrates. Strictly speaking, cats can survive healthily without carbohydrates if they have sufficient animal protein and fat in their diet. However, commercial dry food products for cats often contain large amounts of grain, resulting in carbohydrate levels as high as 35% to 40%. The physical structure of the cat is not good at managing large amounts of carbohydrates, e.g., the chances of developing diabetes and obesity are greatly increased when the cat eats a diet containing large amounts of carbohydrates day after day.
Cats do not like bathing because the water carries away grease from their fur. Although cats like licking their claws and hairs, they are inevitably contaminated with many viruses and are not suitable for frequent bath. The hair is licked and enters the body, so that hair balls are formed, which is not favorable for the health of cats.
Therefore, the design of the cat food with low carbohydrate for protecting the liver and improving the immunity of the cat against virus and the preparation method thereof are necessary.
Disclosure of Invention
The invention aims to provide a cat food for improving immunity and a preparation method thereof, and aims to solve the problems in the background art.
In order to solve the technical problems, the invention provides the following technical scheme: a cat food for improving immunity comprises crab methyl phenethyl alcohol glycoside microspheres, fermented fish meat, compound vitamins, dried mushroom, coarse grain powder, taurine, salt and water.
According to the technical scheme, the crab methallyl alcohol glycoside microcapsule comprises, by mass, 0.02-0.1 part of crab methallyl alcohol glycoside microcapsules, 20-50 parts of fermented fish meat, 0.01-0.5 part of compound vitamins, 1-2 parts of dried mushrooms, 0.01-0.1 part of taurine, 0.1-0.5 part of salt and 5-20 parts of water.
According to the technical scheme, the crab methallyl alcohol glycoside microcapsule is characterized in that liposome of the crab methallyl alcohol glycoside is modified by modified montmorillonite, and then the liposome is prepared into a microsphere structure with sodium alginate and glycine copolymer.
According to the technical scheme, the fermented fish meat is obtained by fermenting lactobacillus plantarum containing a feline interferon (FeIFN-omega) gene.
The preparation method of the immunity-improving cat food comprises the following specific steps:
the method comprises the following steps: stirring and heating the crab toluol glucoside micro-liposome at the speed of 500-800rpm/min to 100 ℃ for 3 minutes, adding the modified montmorillonite, adding the sodium alginate and glycine copolymer, continuously stirring, and stopping stirring for later use after cooling;
step two: stirring fermented fish meat, and mixing with compound vitamins, dried mushroom, taurine, salt, water and the crab toluol glycoside microspheres obtained in the first step to obtain paste;
step three: pouring the obtained pasty mixture into a mold, and putting the mold into an oven for baking;
step four: and cooling the finished cat food and packaging.
According to the technical scheme, the preparation of the copolymer of sodium alginate and glycine comprises the following steps: sodium alginate and glycine were mixed according to 3: 1(w/w), reacting for 96h at 100 ℃ and 120-.
According to the technical scheme, the preparation of the modified montmorillonite comprises the following steps: weighing montmorillonite, dissolving in ultrapure water to prepare 10% montmorillonite suspension, performing ultrasonic treatment for 10min, adding a didodecyldimethylammonium bromide solution with the cation exchange capacity equivalent of 0.5-2 times that of didodecyldimethylammonium bromide, performing ultrasonic-high-speed shearing for 20-30min, performing vacuum freeze drying, grinding, and sieving with a 200-mesh (75 μm) sieve to obtain the modified montmorillonite.
According to the technical scheme, the preparation of the crab toluidine-ethanol glycoside microcapsule comprises the following steps:
dissolving phospholipid, cholesterol and Eriocheir sinensis methyl ethyl alcohol glycoside in anhydrous alcohol, wherein the concentration of phospholipid is 20mg/mL, after the mixture is completely dissolved, rapidly injecting the mixture solution into 2 times volume of phosphate buffer solution (pH6.0, 0.01M) by using an injector, stirring for 10min, carrying out rotary evaporation at 40 ℃ to remove ethanol, adjusting the concentration of phospholipid in liposome to 10mg/mL, and carrying out ultrasonic treatment at 20kHz for 5min (working 10s and intermittent 10s) to prepare the Eriocheir sinensis methyl ethyl alcohol glycoside liposome;
under stirring at 500rpm, the isomorphic modified montmorillonite solution is dripped into the crab toluol glycoside liposome at the dripping speed of 2mL/min, and stirring is continued for 60min after dripping is finished, so as to prepare the modified montmorillonite modified crab toluol glycoside liposome.
Under 500rpm stirring, the copolymer of sodium alginate and glycine is dropped into the modified montmorillonite modified crab toluol glucoside liposome, and the volume ratio is 5: 2, the dropping speed is 2mL/min, and stirring is continued for 60min after the dropping is finished, so as to prepare the crab methyl phenethyl alcohol glycoside microspheres.
According to the technical scheme, the fermented fish meat is obtained by fermenting lactobacillus plantarum containing a feline interferon (FeIFN-omega) gene: adding Lactobacillus plantarum into fish, fermenting at 25-32 deg.C for 48 hr, and inoculating at concentration of 0.02-0.20 g/L.
According to the technical scheme, the preparation of the lactobacillus plantarum containing the feline interferon (FeIFN-omega) gene comprises the following steps: designing and synthesizing primers according to a gene sequence of cat interferon (FeIFN-omega), adding two enzyme cutting sites of Xba I and Hind III at the upstream and downstream of the primers respectively, and adding an upstream primer of FeIFN-omega for: 5'-CCATCTAGAATGTGTGATTTGCCTCAAACTC-3', respectively; downstream primer FeIFN- ω for: 3 '-GTAAGCTTTATTTCTCAGATCTTAATCTTTT-5'; amplifying a cat interferon gene mature region from cat peripheral blood lymphocyte total RNA induced and cultured by ConA by a reverse transcription-polymerase chain reaction technology, and connecting the cat interferon gene mature region to a cloning type vector pMG36 e; the vector pMG36e and FeIFN-omega are subjected to double enzyme digestion by Xba I and Hind III simultaneously, then the two are connected by T4 ligase to obtain a recombinant vector pMG36 e-FeIFN-omega, and finally the vector is electrically transformed into lactobacillus plantarum to obtain the lactobacillus plantarum producing the feline interferon.
Compared with the prior art, the invention has the following beneficial effects: in the invention, the raw materials are mixed,
(1) the cat is a carnivorous animal, the protein required by the body is 5 times of that of a human, the pressure of the kidney can be increased due to excessive protein intake, and the modified sodium alginate is added into cat food to be used as an antioxidant of the cat food, so that free radicals can be eliminated, the renal blood vessels of the cat can be strengthened, and hematuria, urethritis and cystitis caused by glomerular capillary rupture can be prevented; sodium alginate is modified by adopting Maillard reaction to prepare a sodium alginate-glycine copolymer, so that the capacity of removing free radicals of the sodium alginate is improved, the hydrophilicity of the sodium alginate is also improved, the reaction condition is mild, the sodium alginate and the glycine are natural components in food, and the copolymer of the sodium alginate and the glycine is used as a food antioxidant to avoid potential safety hazard caused by chemically synthesizing the antioxidant; in addition, the sodium alginate has the effects of reducing sugar and fat, reducing the influence of high carbohydrate diet on cat blood sugar, protecting cat liver by cooperating with the crab methallyl glucoside, and reducing the burden of high fat diet on cat liver;
(2) the crab methallyl alcohol glycoside has good protection effect on the liver, but has poor stability, can be rapidly degraded under the conditions of high temperature, illumination and high pH value, and is very unfavorable for production and storage; the method comprises the following steps of carrying out intercalation modification on montmorillonite by using a cationic surfactant to ensure that a cationic part is attached to a lamella of the montmorillonite and an organic part is remained between the layers, so that the interlayer spacing of the montmorillonite is increased; meanwhile, the interlayer microenvironment can be improved, so that the montmorillonite is changed from hydrophilicity to hydrophobicity, the surface energy of the montmorillonite is reduced, and the adsorption performance of the montmorillonite on the medicine is improved; modifying liposome of the crab methallyl alcohol glycoside by using modified montmorillonite, wherein the crab methallyl alcohol glycoside is wrapped in the center of the liposome, and the modified montmorillonite is modified on the surface of the liposome through electrostatic interaction; the prepared microsphere is combined with sodium alginate-glycine copolymer to prepare microspheres, and the microsphere structure not only can protect the bioactivity of the crab methallyl alcohol glycoside, but also can play a slow release role, so that the utilization efficiency of the crab methallyl alcohol glycoside is improved; under the digestion of gastric juice, the nondegradable montmorillonite microspheres have a porous structure, so that the hair bulbs in the cat body can be wound on the surfaces of the microspheres, and then the hair bulbs are discharged out of the body;
(3) designing and synthesizing a primer according to a gene sequence of cat interferon (FeIFN-omega) published on Genebank, adding two enzyme cutting sites of Xba I and Hind III on the upstream and downstream of the primer respectively, amplifying the total RNA of cat peripheral blood lymphocytes cultured by ConA induction by a reverse transcription-polymerase chain reaction technology to obtain a mature region of the cat interferon gene, and connecting the mature region to a cloning type vector pMG36 e; the full length of the FeIFN-omega gene is 519bp, and the mature protein of 173 amino acids is coded; the primer sequences are as follows: the upstream primer FeIFN-omega for: 5'-CCATCTAGAATGTGTGATTTGCCTCAAACTC-3', respectively; downstream primer FeIFN- ω for: 3 '-GTAAGCTTTATTTCTCAGATCTTAATCTTTT-5'; the vector pMG36e and FeIFN-omega are subjected to double enzyme digestion by Xba I and Hind III simultaneously, then the two are connected by T4 ligase to obtain a recombinant vector pMG36 e-FeIFN-omega, and finally the vector is electrically transformed into lactobacillus plantarum to obtain lactobacillus plantarum producing the feline interferon, wherein the lactobacillus plantarum is used for fermenting fish meat to obtain fish meat containing the interferon, and the immunity of a cat is improved after the fish meat is eaten.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. 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.
The invention provides the technical scheme that: a cat food for improving immunity comprises crab methyl phenethyl alcohol glycoside microspheres, fermented fish meat, compound vitamins, dried mushroom, coarse grain powder, taurine, salt and water.
According to the technical scheme, the crab methallyl alcohol glycoside microcapsule comprises, by mass, 0.02-0.1 part of crab methallyl alcohol glycoside microcapsules, 20-50 parts of fermented fish meat, 0.01-0.5 part of compound vitamins, 1-2 parts of dried mushrooms, 0.01-0.1 part of taurine, 0.1-0.5 part of salt and 5-20 parts of water.
According to the technical scheme, the crab methallyl alcohol glycoside microcapsule is characterized in that liposome of the crab methallyl alcohol glycoside is modified by modified montmorillonite, and then the liposome is prepared into a microsphere structure with sodium alginate and glycine copolymer.
According to the technical scheme, the fermented fish meat is obtained by fermenting lactobacillus plantarum containing a feline interferon (FeIFN-omega) gene.
The preparation method of the immunity-improving cat food comprises the following specific steps:
the method comprises the following steps: stirring and heating the crab toluol glucoside micro-liposome at the speed of 500-800rpm/min to 100 ℃ for 3 minutes, adding the modified montmorillonite, adding the sodium alginate and glycine copolymer, continuously stirring, and stopping stirring for later use after cooling;
step two: stirring fermented fish meat, and mixing with compound vitamins, dried mushroom, taurine, salt, water and the crab toluol glycoside microspheres obtained in the first step to obtain paste;
step three: pouring the obtained pasty mixture into a mold, and putting the mold into an oven for baking;
step four: and cooling the finished cat food and packaging.
According to the technical scheme, the preparation of the copolymer of sodium alginate and glycine comprises the following steps: sodium alginate and glycine were mixed according to 3: 1(w/w), reacting for 96h at 100 ℃ and 120-.
According to the technical scheme, the preparation of the modified montmorillonite comprises the following steps: weighing montmorillonite, dissolving in ultrapure water to prepare 10% montmorillonite suspension, performing ultrasonic treatment for 10min, adding a didodecyldimethylammonium bromide solution with the cation exchange capacity equivalent of 0.5-2 times that of didodecyldimethylammonium bromide, performing ultrasonic-high-speed shearing for 20-30min, performing vacuum freeze drying, grinding, and sieving with a 200-mesh (75 μm) sieve to obtain the modified montmorillonite.
According to the technical scheme, the preparation of the crab toluidine-ethanol glycoside microcapsule comprises the following steps:
1) dissolving phospholipid, cholesterol and Eriocheir sinensis methyl ethyl alcohol glycoside in anhydrous alcohol, wherein the concentration of phospholipid is 20mg/mL, after the mixture is completely dissolved, rapidly injecting the mixture solution into 2 times volume of phosphate buffer solution (pH6.0, 0.01M) by using an injector, stirring for 10min, carrying out rotary evaporation at 40 ℃ to remove ethanol, adjusting the concentration of phospholipid in liposome to 10mg/mL, and carrying out ultrasonic treatment at 20kHz for 5min (working 10s and intermittent 10s) to prepare the Eriocheir sinensis methyl ethyl alcohol glycoside liposome;
2) under stirring at 500rpm, the isomorphic modified montmorillonite solution is dripped into the crab toluol glycoside liposome at the dripping speed of 2mL/min, and stirring is continued for 60min after dripping is finished, so as to prepare the modified montmorillonite modified crab toluol glycoside liposome.
3) Under 500rpm stirring, the copolymer of sodium alginate and glycine is dropped into the modified montmorillonite modified crab toluol glucoside liposome, and the volume ratio is 5: 2, the dropping speed is 2mL/min, and stirring is continued for 60min after the dropping is finished, so as to prepare the crab methyl phenethyl alcohol glycoside microspheres.
According to the technical scheme, the fermented fish meat is obtained by fermenting lactobacillus plantarum containing a feline interferon (FeIFN-omega) gene: adding Lactobacillus plantarum into fish, fermenting at 25-32 deg.C for 48 hr, and inoculating at concentration of 0.02-0.20 g/L.
According to the technical scheme, the preparation of the lactobacillus plantarum containing the feline interferon (FeIFN-omega) gene comprises the following steps: designing and synthesizing primers according to a gene sequence of cat interferon (FeIFN-omega), adding two enzyme cutting sites of Xba I and Hind III at the upstream and downstream of the primers respectively, and adding an upstream primer of FeIFN-omega for: 5'-CCATCTAGAATGTGTGATTTGCCTCAAACTC-3', respectively; downstream primer FeIFN- ω for: 3 '-GTAAGCTTTATTTCTCAGATCTTAATCTTTT-5'; amplifying a cat interferon gene mature region from cat peripheral blood lymphocyte total RNA induced and cultured by ConA by a reverse transcription-polymerase chain reaction technology, and connecting the cat interferon gene mature region to a cloning type vector pMG36 e; the vector pMG36e and FeIFN-omega are subjected to double enzyme digestion by Xba I and Hind III simultaneously, then the two are connected by T4 ligase to obtain a recombinant vector pMG36 e-FeIFN-omega, and finally the vector is electrically transformed into lactobacillus plantarum to obtain the lactobacillus plantarum producing the feline interferon.
Example 1
A cat food for improving immunity comprises crab methyl phenethyl alcohol glycoside microspheres, fermented fish meat, compound vitamins, dried mushroom, coarse grain powder, taurine, salt and water.
According to the technical scheme, the crab methallyl alcohol glycoside microcapsule comprises, by mass, 0.1 part of crab methallyl alcohol glycoside microcapsules, 30 parts of fermented fish meat, 0.01 part of compound vitamins, 1 part of dried mushrooms, 0.03 part of taurine, 0.1 part of salt and 15 parts of water.
According to the technical scheme, the crab methallyl alcohol glycoside microcapsule is characterized in that liposome of the crab methallyl alcohol glycoside is modified by modified montmorillonite, and then the liposome is prepared into a microsphere structure with sodium alginate and glycine copolymer.
According to the technical scheme, the fermented fish meat is obtained by fermenting lactobacillus plantarum containing a feline interferon (FeIFN-omega) gene.
The preparation method of the immunity-improving cat food comprises the following specific steps:
the method comprises the following steps: stirring and heating the crab toluol glucoside liposome at 500rpm/min to 100 ℃ for 3 minutes, adding the modified montmorillonite, adding the sodium alginate-glycine copolymer, continuously stirring, and stopping stirring for later use after cooling;
step two: stirring fermented fish meat, and mixing with compound vitamins, dried mushroom, taurine, salt, water and the crab toluol glycoside microspheres obtained in the first step to obtain paste;
step three: pouring the obtained pasty mixture into a mold, and putting the mold into an oven for baking;
step four: and cooling the finished cat food and packaging.
According to the technical scheme, the preparation of the copolymer of sodium alginate and glycine comprises the following steps: sodium alginate and glycine were mixed according to 3: 1(w/w), stirring at 100 ℃ and 180rpm/min, and reacting for 96 hours to form the copolymer of sodium alginate and glycine.
According to the technical scheme, the preparation of the modified montmorillonite comprises the following steps: weighing montmorillonite, dissolving in ultrapure water to prepare 10% montmorillonite suspension, performing ultrasonic treatment for 10min, adding a didodecyldimethylammonium bromide solution with the cation exchange capacity equivalent of 0.5-2 times that of didodecyldimethylammonium bromide, performing ultrasonic-high-speed shearing for 30min, performing vacuum freeze drying, grinding, and sieving with a 200-mesh (75 μm) sieve to obtain the modified montmorillonite.
According to the technical scheme, the preparation of the crab toluidine-ethanol glycoside microcapsule comprises the following steps:
1) dissolving phospholipid, cholesterol and Eriocheir sinensis methyl ethyl alcohol glycoside in anhydrous alcohol, wherein the concentration of phospholipid is 20mg/mL, after the mixture is completely dissolved, rapidly injecting the mixture solution into 2 times volume of phosphate buffer solution (pH6.0, 0.01M) by using an injector, stirring for 10min, carrying out rotary evaporation at 40 ℃ to remove ethanol, adjusting the concentration of phospholipid in liposome to 10mg/mL, and carrying out ultrasonic treatment at 20kHz for 5min (working 10s and intermittent 10s) to prepare the Eriocheir sinensis methyl ethyl alcohol glycoside liposome;
2) under stirring at 500rpm, the isomorphic modified montmorillonite solution is dripped into the crab toluol glycoside liposome at the dripping speed of 2mL/min, and stirring is continued for 60min after dripping is finished, so as to prepare the modified montmorillonite modified crab toluol glycoside liposome.
3) Under 500rpm stirring, the copolymer of sodium alginate and glycine is dropped into the modified montmorillonite modified crab toluol glucoside liposome, and the volume ratio is 5: 2, the dropping speed is 2mL/min, and stirring is continued for 60min after the dropping is finished, so as to prepare the crab methyl phenethyl alcohol glycoside microspheres.
According to the technical scheme, the fermented fish meat is obtained by fermenting lactobacillus plantarum containing a feline interferon (FeIFN-omega) gene: adding Lactobacillus plantarum into fish, fermenting at 25-32 deg.C for 48 hr, and inoculating at concentration of 0.10 g/L.
According to the technical scheme, the preparation of the lactobacillus plantarum containing the feline interferon (FeIFN-omega) gene comprises the following steps: designing and synthesizing primers according to a gene sequence of cat interferon (FeIFN-omega), adding two enzyme cutting sites of Xba I and Hind III at the upstream and downstream of the primers respectively, and adding an upstream primer of FeIFN-omega for: 5'-CCATCTAGAATGTGTGATTTGCCTCAAACTC-3', respectively; downstream primer FeIFN- ω for: 3 '-GTAAGCTTTATTTCTCAGATCTTAATCTTTT-5'; amplifying a cat interferon gene mature region from cat peripheral blood lymphocyte total RNA induced and cultured by ConA by a reverse transcription-polymerase chain reaction technology, and connecting the cat interferon gene mature region to a cloning type vector pMG36 e; the vector pMG36e and FeIFN-omega are subjected to double enzyme digestion by Xba I and Hind III simultaneously, then the two are connected by T4 ligase to obtain a recombinant vector pMG36 e-FeIFN-omega, and finally the vector is electrically transformed into lactobacillus plantarum to obtain the lactobacillus plantarum producing the feline interferon.
Example 2
A cat food for improving immunity comprises crab methyl phenethyl alcohol glycoside microspheres, fermented fish meat, compound vitamins, dried mushroom, coarse grain powder, taurine, salt and water.
According to the technical scheme, the crab methallyl alcohol glycoside microcapsule comprises, by mass, 0.1 part of crab methallyl alcohol glycoside microcapsules, 20 parts of fermented fish meat, 0.01 part of compound vitamins, 1 part of dried mushrooms, 0.02 part of taurine, 0.1 part of salt and 12 parts of water.
According to the technical scheme, the crab methallyl alcohol glycoside microcapsule is characterized in that liposome of the crab methallyl alcohol glycoside is modified by modified montmorillonite, and then the liposome is prepared into a microsphere structure with sodium alginate and glycine copolymer.
According to the technical scheme, the fermented fish meat is obtained by fermenting lactobacillus plantarum containing a feline interferon (FeIFN-omega) gene.
The preparation method of the immunity-improving cat food comprises the following specific steps:
the method comprises the following steps: stirring and heating the crab toluol glucoside liposome at 800rpm/min to 100 ℃ for 3 minutes, adding the modified montmorillonite, adding the sodium alginate-glycine copolymer, continuously stirring, and stopping stirring for later use after cooling;
step two: stirring fermented fish meat, and mixing with compound vitamins, dried mushroom, taurine, salt, water and the crab toluol glycoside microspheres obtained in the first step to obtain paste;
step three: pouring the obtained pasty mixture into a mold, and putting the mold into an oven for baking;
step four: and cooling the finished cat food and packaging.
According to the technical scheme, the preparation of the copolymer of sodium alginate and glycine comprises the following steps: sodium alginate and glycine were mixed according to 3: 1(w/w), stirring at 100 ℃ and 200rpm/min, and reacting for 96 hours to form the copolymer of sodium alginate and glycine.
According to the technical scheme, the preparation of the modified montmorillonite comprises the following steps: weighing montmorillonite, dissolving in ultrapure water to prepare 10% montmorillonite suspension, performing ultrasonic treatment for 10min, adding a didodecyldimethylammonium bromide solution with the cation exchange capacity equivalent of 0.5 times that of didodecyldimethylammonium bromide, performing ultrasonic-high-speed shearing for 20-30min, performing vacuum freeze drying, grinding, and sieving with a 200-mesh (75 μm) sieve to obtain the modified montmorillonite.
According to the technical scheme, the preparation of the crab toluidine-ethanol glycoside microcapsule comprises the following steps:
1) dissolving phospholipid, cholesterol and Eriocheir sinensis methyl ethyl alcohol glycoside in anhydrous alcohol, wherein the concentration of phospholipid is 20mg/mL, after the mixture is completely dissolved, rapidly injecting the mixture solution into 2 times volume of phosphate buffer solution (pH6.0, 0.01M) by using an injector, stirring for 10min, carrying out rotary evaporation at 40 ℃ to remove ethanol, adjusting the concentration of phospholipid in liposome to 10mg/mL, and carrying out ultrasonic treatment at 20kHz for 5min (working 10s and intermittent 10s) to prepare the Eriocheir sinensis methyl ethyl alcohol glycoside liposome;
2) under stirring at 500rpm, the isomorphic modified montmorillonite solution is dripped into the crab toluol glycoside liposome at the dripping speed of 2mL/min, and stirring is continued for 60min after dripping is finished, so as to prepare the modified montmorillonite modified crab toluol glycoside liposome.
3) Under 500rpm stirring, the copolymer of sodium alginate and glycine is dropped into the modified montmorillonite modified crab toluol glucoside liposome, and the volume ratio is 5: 2, the dropping speed is 2mL/min, and stirring is continued for 60min after the dropping is finished, so as to prepare the crab methyl phenethyl alcohol glycoside microspheres.
According to the technical scheme, the fermented fish meat is obtained by fermenting lactobacillus plantarum containing a feline interferon (FeIFN-omega) gene: adding Lactobacillus plantarum into fish, fermenting at 30 deg.C for 48 hr, and inoculating at a concentration of 0.20 g/L.
According to the technical scheme, the preparation of the lactobacillus plantarum containing the feline interferon (FeIFN-omega) gene comprises the following steps: designing and synthesizing primers according to a gene sequence of cat interferon (FeIFN-omega), adding two enzyme cutting sites of Xba I and Hind III at the upstream and downstream of the primers respectively, and adding an upstream primer of FeIFN-omega for: 5'-CCATCTAGAATGTGTGATTTGCCTCAAACTC-3', respectively; downstream primer FeIFN- ω for: 3 '-GTAAGCTTTATTTCTCAGATCTTAATCTTTT-5'; amplifying a cat interferon gene mature region from cat peripheral blood lymphocyte total RNA induced and cultured by ConA by a reverse transcription-polymerase chain reaction technology, and connecting the cat interferon gene mature region to a cloning type vector pMG36 e; the vector pMG36e and FeIFN-omega are subjected to double enzyme digestion by Xba I and Hind III simultaneously, then the two are connected by T4 ligase to obtain a recombinant vector pMG36 e-FeIFN-omega, and finally the vector is electrically transformed into lactobacillus plantarum to obtain the lactobacillus plantarum producing the feline interferon.
In order to verify the beneficial effects of the invention, the following tests are specially carried out:
test 1. scavenging ability of sodium alginate-glycine copolymer to free radical
The hydroxyl radical is removed by Fenton reaction
Taking 3 colorimetric tubes with the volume of 10mL, respectively and sequentially adding 1.0mL7.5mmol/L ferrous sulfate iron solution, 1.0mL7.5mmol/L salicylic acid solution and 1.0mL7.5mmol/L hydrogen peroxide solution, sequentially adding 1.0mL sodium alginate-glycine copolymer into the No. 2 colorimetric tube, fixing the volume to the scale with deionized water, heating in a water bath for 1h, sequentially adding 1.0mL sodium alginate into the No. 3 colorimetric tube, fixing the volume to the scale with deionized water, and heating in the water bath for 1 h. The absorbance values were measured at 510nm wavelength and the clearance was calculated.
Clearance rate equation: clearance (%) ═ a0-As)/A0*100％
In the formula: a. the0-no absorbance value of the test substance is added; as-absorbance after addition to assay
| Numbering | Light absorption value (510nm) | Percent removal (%) |
| Sodium alginate-glycine copolymer | 0.083 | 88.6 |
| Sodium alginate | 0.092 | 56.4 |
TABLE 1 statistics of sodium alginate-glycine copolymer on radical clearance
As can be seen from the test results, the sodium alginate-glycine copolymer has good ability to scavenge free radicals, and has stronger ability to scavenge free radicals than pure sodium alginate.
Experiment 2. experiment of degrading fatty liver fat with cat food of the present invention
2.1 preparation of fatty liver model
After 7 days of adaptive feeding all rats were randomized into 2 groups, respectively a high lipid model group (HF, 20) and a blank control group (NC, Normal control, 10). HF rats were fed with high-fat diet (78.5% conventional diet + 0.5% sodium cholate + 1% cholesterol + 10% egg yolk powder + 10% lard), NC group rats were fed with conventional diet with free access to water, 12h light/12 h dark at 23 ℃. + -. 2 ℃. The molding time was 14 d.
2.2 animal grouping and test treatment
The high-fat model group was randomly divided into two groups of 10 animals, one group (HF1) was fed with cat food for 8 days, and the other group (HF2) was fed with regular diet for 6 weeks as a blank control group (NC), and the feed quality was the same. All rats were sacrificed and livers were weighed with the specific values as in table 1:
| group of | Average liver weight (g) | Average body weight (g) | Liver to body weight ratio (%) |
| Blank control group (NC) | 0.841 | 352 | 0.239 |
| High fat model group (HF1) | 0.899 | 367 | 0.245 |
| Two groups of high fat model (HF2) | 0.974 | 373 | 0.261 |
TABLE 1 statistical table of rat liver and body weight
As can be seen from table 1, the weight of the rats fed with cat food in the high-fat model group was not significantly reduced, but the weight of the liver was significantly reduced, and the mass ratio of the weight of the liver to the weight of the body was close to that of the blank control, which indicates that the cat food had a good effect of improving the degradation of fatty liver fat in the rats.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
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 changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A cat food for improving immunity comprises crab methyl phenethyl alcohol glycoside microspheres, fermented fish meat, compound vitamins, dried mushroom, coarse grain powder, taurine, salt and water.
2. The immunity-enhancing cat food as claimed in claim 1, wherein the immunity-enhancing cat food comprises the following components: the crab methallyl alcohol glycoside microcapsule comprises, by mass, 0.02-0.1 part of crab methallyl alcohol glycoside microcapsules, 20-50 parts of fermented fish meat, 0.01-0.5 part of compound vitamins, 1-2 parts of dried mushrooms, 0.01-0.1 part of taurine, 0.1-0.5 part of salt and 5-20 parts of water.
3. The immunity-enhancing cat food as claimed in claim 1, wherein the immunity-enhancing cat food comprises the following components: the crab methallyl alcohol glycoside microcapsule is characterized in that liposome of the crab methallyl alcohol glycoside is modified by modified montmorillonite, and then the liposome is prepared into a microsphere structure with sodium alginate and glycine copolymer.
4. The immunity-enhancing cat food as claimed in claim 1, wherein the immunity-enhancing cat food comprises the following components: the fermented fish meat is obtained by fermenting lactobacillus plantarum containing a feline interferon (FeIFN-omega) gene.
5. The preparation method of the immunity-improving cat food comprises the following specific steps:
the method comprises the following steps: stirring and heating the crab toluol glucoside micro-liposome at the speed of 500-800rpm/min to 100 ℃ for 3 minutes, adding the modified montmorillonite, adding the sodium alginate and glycine copolymer, continuously stirring, and stopping stirring for later use after cooling;
step two: stirring fermented fish meat, and mixing with compound vitamins, dried mushroom, taurine, salt, water and the crab toluol glycoside microspheres obtained in the first step to obtain paste;
step three: pouring the obtained pasty mixture into a mold, and putting the mold into an oven for baking;
step four: and cooling the finished cat food and packaging.
6. The preparation method of the immunity-improving cat food according to claim 5, which is characterized by comprising the following steps: the preparation method of the copolymer of sodium alginate and glycine comprises the following steps: sodium alginate and glycine were mixed according to 3: 1(w/w), reacting for 96h at 100 ℃ and 120-.
7. The preparation method of the immunity-improving cat food according to claim 5, which is characterized by comprising the following steps: preparing the modified montmorillonite: weighing montmorillonite, dissolving in ultrapure water to prepare 10% montmorillonite suspension, performing ultrasonic treatment for 10min, adding a didodecyldimethylammonium bromide solution with the cation exchange capacity equivalent of 0.5-2 times that of didodecyldimethylammonium bromide, performing ultrasonic-high-speed shearing for 20-30min, performing vacuum freeze drying, grinding, and sieving with a 200-mesh (75 μm) sieve to obtain the modified montmorillonite.
8. The preparation method of the immunity-improving cat food according to claim 5, which is characterized by comprising the following steps: the preparation method of the crab methallyl alcohol glycoside microcapsule comprises the following steps:
1) dissolving phospholipid, cholesterol and Eriocheir sinensis methyl ethyl alcohol glycoside in anhydrous alcohol, wherein the concentration of phospholipid is 20mg/mL, after the mixture is completely dissolved, rapidly injecting the mixture solution into 2 times volume of phosphate buffer solution (pH6.0, 0.01M) by using an injector, stirring for 10min, carrying out rotary evaporation at 40 ℃ to remove ethanol, adjusting the concentration of phospholipid in liposome to 10mg/mL, and carrying out ultrasonic treatment at 20kHz for 5min (working 10s and intermittent 10s) to prepare the Eriocheir sinensis methyl ethyl alcohol glycoside liposome;
2) under stirring at 500rpm, the isomorphic modified montmorillonite solution is dripped into the crab toluol glycoside liposome at the dripping speed of 2mL/min, and stirring is continued for 60min after dripping is finished, so as to prepare the modified montmorillonite modified crab toluol glycoside liposome.
3) Under 500rpm stirring, the copolymer of sodium alginate and glycine is dropped into the modified montmorillonite modified crab toluol glucoside liposome, and the volume ratio is 5: 2, the dropping speed is 2mL/min, and stirring is continued for 60min after the dropping is finished, so as to prepare the crab methyl phenethyl alcohol glycoside microspheres.
9. The preparation method of the immunity-improving cat food according to claim 5, which is characterized by comprising the following steps: the fermented fish meat is obtained by fermenting lactobacillus plantarum containing a feline interferon (FeIFN-omega) gene: adding Lactobacillus plantarum into fish, fermenting at 25-32 deg.C for 48 hr, and inoculating at concentration of 0.02-0.20 g/L.
10. The preparation method of the immunity-improving cat food according to claim 9, which is characterized by comprising the following steps: the preparation of the lactobacillus plantarum containing the feline interferon (FeIFN-omega) gene comprises the following steps: designing and synthesizing primers according to a gene sequence of cat interferon (FeIFN-omega), adding two enzyme cutting sites of Xba I and Hind III at the upstream and downstream of the primers respectively, and adding an upstream primer of FeIFN-omega for: 5'-CCATCTAGAATGTGTGATTTGCCTCAAACTC-3', respectively; downstream primer FeIFN- ω for: 3 '-GTAAGCTTTATTTCTCAGATCTTAATCTTTT-5'; amplifying a cat interferon gene mature region from cat peripheral blood lymphocyte total RNA induced and cultured by ConA by a reverse transcription-polymerase chain reaction technology, and connecting the cat interferon gene mature region to a cloning type vector pMG36 e; the vector pMG36e and FeIFN-omega are subjected to double enzyme digestion by Xba I and Hind III simultaneously, then the two are connected by T4 ligase to obtain a recombinant vector pMG36 e-FeIFN-omega, and finally the vector is electrically transformed into lactobacillus plantarum to obtain the lactobacillus plantarum producing the feline interferon.
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