CN114916626B - Probiotic fish feed containing traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide and application thereof - Google Patents
Probiotic fish feed containing traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide and application thereof Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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- Engineering & Computer Science (AREA)
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- Biotechnology (AREA)
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- Mycology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
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- Insects & Arthropods (AREA)
- Marine Sciences & Fisheries (AREA)
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- Fodder In General (AREA)
Abstract
The invention relates to a probiotic fish feed containing traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide, and a preparation method and application thereof. According to the invention, the traditional Chinese medicine aqueous extract alcohol sediment and the traditional Chinese medicine residues are deeply utilized, related active ingredient polysaccharide is extracted and combined with mulberry leaves and further processed to obtain the prebiotic composition containing traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide and the prebiotic functional fish feed, which can effectively improve the meat quality of fish, promote the growth of fish, fully realize the purposes of reutilizing wastes and changing wastes into valuables, and effectively reduce the pollution of the wastes to the environment in the traditional Chinese medicine preparation process.
Description
Technical Field
The invention belongs to the field of feeds, and particularly relates to a probiotic fish feed containing traditional Chinese medicine oligosaccharides and mulberry leaf oligosaccharides, and a preparation method and application thereof.
Background
The Chinese medicine is a large country for producing and using Chinese medicines, and has a great variety of Chinese medicines, huge yield and long history. Along with the rapid development of the Chinese traditional medicine processing industry, the yield of the Chinese traditional medicine solid slag is continuously increased. According to statistics, the traditional Chinese medicine solid slag generated in China reaches 7000 ten thousand tons each year, and is limited by the limit of an extraction process and technology, a large amount of medicinal components, crude fibers, polysaccharide, starch and unknown growth promoting factors still remain in the traditional Chinese medicine solid slag, and the traditional Chinese medicine solid slag treatment mode is methods of incineration, backfilling, fixed area stacking and the like.
When Chinese traditional medicine enterprises produce granular products, in order to ensure uniform and stable product quality, the Chinese traditional medicine materials after water extraction are required to be subjected to alcohol precipitation treatment, supernatant liquid after alcohol precipitation is used as a raw material for producing the Chinese traditional medicine granules, and liquid alcohol precipitation is waste, but contains abundant polysaccharide and flavonoid substances. In actual production, the liquid waste is often directly discharged, and the ecological environment is greatly damaged.
The mulberry is a woody plant with fallen leaves of Sang Keduo years, has strong stress resistance and has a large planting area in China. Mulberry leaves are the main product of mulberry, can be picked many times each year, and have extremely rich yield. The mulberry leaves are rich in nutritional elements such as protein, vitamins and minerals, and bioactive components such as mulberry leaf polysaccharide, mulberry leaf flavone and alkaloid, have various physiological functions such as blood sugar reduction, anti-inflammatory and antivirus, can improve the immunity of aquatic livestock and poultry animals, can improve the muscle quality of the animals, improve the meat quality and flavor, and have a certain effect, thus having great development potential in the aspect of healthy cultivation.
Although the solid waste residues and the liquid alcohol sediment of the traditional Chinese medicines contain components beneficial to animals, the application of the solid waste residues and the liquid alcohol sediment to the production of fish feed is limited by factors such as anti-nutritional factors, poor palatability and the like, so that the growth of animals is limited, and the direct addition of the solid waste residues and the liquid alcohol sediment into the animal feed inevitably causes a certain harm to the animals, and serious death even occurs. Therefore, the fish feed is prepared after proper treatment and auxiliary materials are matched, so that the purpose of changing waste into valuable is achieved by the traditional Chinese medicine residues. Meanwhile, the prior art does not adopt Chinese medicinal residues and mulberry leaves as raw materials to prepare the fermented fish feed for improving the meat quality of fish.
Disclosure of Invention
The invention aims to solve the problems in the prior art, and recycle the water-extracted alcohol matters, the traditional Chinese medicine residues and other wastes generated in the traditional Chinese medicine production, extract the effective active ingredients in the water-extracted alcohol matters, and deeply process the effective active ingredients together with the mulberry leaves as raw materials to prepare the feed with the functions of improving the fish quality and promoting the fish growth, so as to realize the purpose of recycling the wastes and reduce the damage of the traditional Chinese medicine wastes to the environment.
To achieve the above object, the present invention is achieved by the following means:
the first aspect of the present invention provides a method of preparing a prebiotic composition comprising the steps of:
(1) Pulverizing folium Mori, sieving, extracting with water, concentrating, precipitating with ethanol, recovering ethanol, and collecting precipitate to obtain folium Mori polysaccharide;
(2) Collecting concentrated liquid medicine to extract alcohol precipitate, recovering ethanol, and measuring and diluting the concentration of polysaccharide in the concentrated liquid medicine; meanwhile, the concentration of the mulberry leaf polysaccharide obtained in the step (1) is measured and diluted;
(3) Mixing the traditional Chinese medicine polysaccharide diluted in the step (2) with mulberry leaf polysaccharide, and performing ultrasonic degradation; the purpose of sonication is to reduce polysaccharide viscosity;
(4) Performing enzymolysis treatment on the polysaccharide mixture subjected to ultrasonic degradation in the step (3);
(5) Adding pretreated macroporous resin into the solution subjected to enzymolysis in the step (4) for oscillation adsorption, and then filtering to remove the macroporous resin to obtain the traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition after flavone removal.
Preferably, the screen holes for crushing and sieving the mulberry leaves in the step (1) are 10 meshes.
Preferably, the temperature of the water extraction in the step (1) is 70 ℃, and the time of the water extraction is 3 hours;
preferably, the concentration in the step (1) means that the liquid volume is 1/2 of that before concentration.
Preferably, in the step (1), the concentration of the ethanol is 95%, the volume of the ethanol is 2 times that of the concentrated solution, and the time of the ethanol precipitation treatment is 24 hours.
Preferably, the water-extracted alcohol precipitate of the traditional Chinese medicine in the step (2) includes, but is not limited to, one or more of a summer Sang Jushui water-extracted alcohol precipitate produced by Guangzhou white cloud mountain star (pharmaceutical industry) and a pediatric seven-star tea water-extracted alcohol precipitate produced by Guangzhou Wang Lou Ji pharmaceutical industry, a Wang Laojia water-extracted alcohol precipitate, and a radix Isatidis water-extracted alcohol precipitate produced by Guangzhou Xiangxue pharmaceutical industry.
Preferably, the concentration measurement method of the traditional Chinese medicine polysaccharide in the step (2) is to measure by using a sulfuric acid-phenol method; the concentration of the traditional Chinese medicine polysaccharide after dilution is 0.1-100mg/mL.
Preferably, the concentration measuring method of the mulberry leaf polysaccharide in the step (2) is to measure by using a sulfuric acid-phenol method; the concentration of the mulberry leaf polysaccharide after dilution is 2-20mg/mL.
Preferably, the conditions of the ultrasonication in step (3) are: the ultrasonic power is 300W, and the degradation time is 2-20min.
Preferably, the enzymolysis treatment in the step (4) specifically comprises: the enzyme is firstly subjected to enzymolysis for 1-2 hours at 30-55 ℃ by using hemicellulase, then is subjected to enzymolysis for 0.5-1 hour at 30-60 ℃ by using pectase, and finally is subjected to enzymolysis for 2-4 hours at 30-50 ℃ by using saccharifying enzyme, and is subjected to enzyme deactivation by heating for 20 minutes at 85 ℃.
Preferably, the macroporous resin in step (5) is selected from one or more of HDP600, YKDH-9, YKDH-2, NKA and H107; the dosage of the macroporous resin is 0.01g/mL.
Preferably, the oscillating adsorption in the step (5) is specifically: the temperature is 40 ℃, the adsorption time is 3 hours, and the oscillating rotation speed is 120r/min.
In a second aspect the present invention provides a prebiotic composition prepared according to the preparation method described above.
The third aspect of the invention provides a prebiotic functional fish feed, comprising a functional feed and a basic fish feed, wherein the functional feed comprises traditional Chinese medicine residues, mulberry leaves and a prebiotic composition prepared according to the method.
Preferably, the dosage of the prebiotic composition is 0.15-15% of the total mass of the traditional Chinese medicine residues and the mulberry leaves.
Preferably, the dosage ratio of the functional feed to the basal fish feed is 3:7.
Preferably, the Chinese medicine dregs include, but are not limited to, one or more of Xiasangju decoction dregs produced by Guangzhou white cloud mountain star (pharmaceutical industry) share limited company, pediatric Qixing tea dregs produced by Guangzhou Wang Lou Ji pharmaceutical industry share limited company, wang Lou Ji dregs, and Isatis root dregs produced by Guangzhou Fang Xueu pharmaceutical industry share limited company.
Preferably, the basic fish feed is a fish feed commonly used in the art and is commercially available, for example, the basic fish feed comprises one or more of rice bran, wheat bran, bean cake, fish meal, yeast powder, corn meal, straw powder, silkworm chrysalis powder, wheat middling, bone meal, rapeseed cake powder, barley flour and edible salt, and the skilled person is fully capable of adaptively selecting and adjusting the ingredients and the dosage proportion in the basic fish feed according to the type of the fish to be bred.
The fourth aspect of the invention provides a method for preparing prebiotic functional fish feed, comprising the following steps:
(1) Oven drying the residue and folium Mori, pulverizing, and sieving;
(2) Adding the prebiotic composition for fermentation;
(3) After fermentation is completed, drying treatment is carried out;
(4) Crushing and sieving the dried mixture to obtain functional feed;
(5) Adding basic feed into the functional feed, and mixing.
Preferably, the mesh size of the crushing and sieving in the step (1) is 10 mesh.
Preferably, the fermentation in step (2) is specifically: adding 0.1-10% of traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic mixture according to the total mass, inoculating Aspergillus niger, and fermenting at 30-35 ℃ for 1-3d; then inoculating bacillus and fermenting for 1d at 30-37 ℃; finally, adding 0.01-5% of prebiotic composition according to the total mass, inoculating lactobacillus and yeast, and fermenting at 37-45deg.C for 1-2d.
Preferably, the drying in the step (3) is specifically: drying at 60-85deg.C for 1-4 hr.
Preferably, the mesh size of the screen in step (4) is 20 mesh.
Preferably, the ratio of the functional feed to the basal fish feed in step (5) is 3:7.
It should be understood that, in the context of the present invention, the water-extracted alcohol precipitate and/or the residue of the traditional Chinese medicine may be selected from the waste materials formed by the conventional traditional Chinese medicine enterprises in the daily production process, for example, one or more of the water-extracted alcohol precipitate and residue of summer Sang Jushui from Guangzhou white cloud mountain star (pharmaceutical industry) and the residue and residue of radix Isatidis from Guangzhou Wang Lou Ji pharmaceutical industry, inc. unless otherwise specified. The above listed manufacturers and types of traditional Chinese medicines are only examples and do not limit the practice of the present invention.
Compared with the prior art, the invention has the following beneficial effects:
according to the invention, the traditional Chinese medicine aqueous extract alcohol sediment and the traditional Chinese medicine residues are deeply utilized, related active ingredient polysaccharide is extracted and combined with mulberry leaves and further processed to obtain the prebiotic composition containing traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide and the prebiotic functional fish feed, which can effectively improve the meat quality of fish, promote the growth of fish, fully realize the purposes of reutilizing wastes and changing wastes into valuables, and effectively reduce the pollution of the wastes to the environment in the traditional Chinese medicine preparation process.
Detailed Description
In order to make the objects, technical solutions and effects of the present invention more clear and clear, the present invention will be described in further detail with reference to examples. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention. Unless defined otherwise, all technical and scientific terms used hereinafter have the same meaning as commonly understood by one of ordinary skill in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the scope of the present invention.
Example 1
A prebiotic composition, the method of preparation comprising the steps of:
(1) Pulverizing folium Mori, sieving with 10 mesh sieve, extracting with 70deg.C water for 3 hr, concentrating the liquid to 1/2, precipitating with 2 times of 95% ethanol for 24 hr, recovering ethanol, and collecting precipitate to obtain folium Mori polysaccharide;
(2) Collecting summer Sang Jushui ethanol precipitate produced by Guangzhou white cloud mountain star (pharmaceutical industry) and recovering ethanol, and measuring and diluting the polysaccharide concentration of the traditional Chinese medicine by sulfuric acid-phenol method to reach polysaccharide concentration of 50mg/mL; meanwhile, the concentration of the mulberry leaf polysaccharide obtained in the step (1) is measured and diluted by a sulfuric acid-phenol method, so that the concentration of the mulberry leaf polysaccharide reaches 10mg/mL;
(3) Mixing the traditional Chinese medicine polysaccharide diluted in the step (2) with mulberry leaf polysaccharide, and performing ultrasonic degradation, wherein the ultrasonic power is 300W, and the degradation time is 10min; the purpose of sonication is to reduce polysaccharide viscosity;
(4) Performing enzymolysis treatment on the polysaccharide mixture subjected to ultrasonic degradation in the step (3), wherein the enzymolysis treatment comprises the following specific steps: firstly, performing enzymolysis for 1.5 hours at 45 ℃ by using hemicellulase, then performing enzymolysis for 0.8 hour at 50 ℃ by using pectase, and finally performing enzymolysis for 3 hours at 40 ℃ by using saccharifying enzyme, and heating for 20 minutes at 85 ℃ to inactivate enzymes;
(5) Adding pretreated HDP600 macroporous resin into the solution subjected to enzymolysis in the step (4), and carrying out vibration adsorption for 3 hours at 40 ℃ and 120r/min, wherein the dosage of the macroporous resin is 0.01g/mL; and filtering to remove macroporous resin to obtain the traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition after flavone removal.
Further, the preparation of the functional fish feed by using the prebiotic composition comprises the following steps:
(1) Oven drying residue of XIASANGJU (produced by Guangzhou white cloud mountain Star (pharmaceutical industry) stock, and pulverizing and sieving with 10 mesh sieve;
(2) Adding 5% by weight of the prebiotic composition for fermentation; the fermentation steps are as follows: inoculating Aspergillus niger and fermenting at 32deg.C for 2d; then inoculating bacillus and fermenting for 1d at 34 ℃; finally, adding 1% of traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition according to the total mass, inoculating lactobacillus and yeast, and fermenting for 1d at 40 ℃;
(3) After fermentation is completed, drying for 3 hours at 70 ℃;
(4) Crushing the dried mixture, and sieving the crushed mixture with a 20-mesh sieve to obtain functional feed;
(5) Uniformly mixing the functional feed and the basic fish feed according to the mass ratio of 3:7 to obtain the feed; the basic fish feed consists of the following components in percentage by mass: 40% of rice bran, 38% of wheat bran, 10% of bean cake, 10% of fish meal and 2% of yeast powder.
Example 2
A prebiotic mixture, the method of preparation comprising the steps of:
(1) Pulverizing folium Mori, sieving with 10 mesh sieve, extracting with 70deg.C water for 3 hr, concentrating the liquid to 1/2, precipitating with 2 times of 95% ethanol for 24 hr, recovering ethanol, and collecting precipitate to obtain folium Mori polysaccharide;
(2) Collecting radix Isatidis water extract and alcohol precipitate produced by Guangzhou city chikungunya pharmaceutical Co., ltd, recovering ethanol, measuring the polysaccharide concentration of the Chinese medicinal materials by sulfuric acid-phenol method, and diluting to make the polysaccharide concentration reach 35mg/mL; meanwhile, the concentration of the mulberry leaf polysaccharide obtained in the step (1) is measured and diluted by a sulfuric acid-phenol method, so that the concentration of the mulberry leaf polysaccharide reaches 15mg/mL;
(3) Mixing the traditional Chinese medicine polysaccharide diluted in the step (2) with mulberry leaf polysaccharide, and performing ultrasonic degradation, wherein the ultrasonic power is 300W, and the degradation time is 8min; the purpose of sonication is to reduce polysaccharide viscosity;
(4) Performing enzymolysis treatment on the polysaccharide mixture subjected to ultrasonic degradation in the step (3), wherein the enzymolysis treatment comprises the following specific steps: firstly, performing enzymolysis for 1h at 55 ℃ by using hemicellulase, then performing enzymolysis for 0.5h at 60 ℃ by using pectase, finally performing enzymolysis for 2h at 50 ℃ by using saccharifying enzyme, and heating for 20min at 85 ℃ to inactivate enzyme;
(5) Adding pretreated NKA macroporous resin into the solution subjected to enzymolysis in the step (4), and performing vibration adsorption for 3 hours at 40 ℃ and 120r/min, wherein the dosage of the macroporous resin is 0.01g/mL; and filtering to remove macroporous resin to obtain the traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition after flavone removal.
Further, the preparation of the functional fish feed by using the prebiotic composition comprises the following steps:
(1) Drying and crushing radix isatidis dregs produced by Guangzhou city chiffon pharmaceutical Co., ltd, and sieving with a 10-mesh sieve;
(2) Adding 0.8% wt of the prebiotic composition for fermentation; the fermentation steps are as follows: inoculating Aspergillus niger and fermenting at 30deg.C for 1d; then inoculating bacillus and fermenting for 1d at 30 ℃; finally, adding 5% of traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition according to the total mass, inoculating lactobacillus and yeast, and fermenting for 2d at 45 ℃;
(3) After fermentation is completed, drying for 1h at 85 ℃;
(4) Crushing the dried mixture, and sieving the crushed mixture with a 20-mesh sieve to obtain functional feed;
(5) Uniformly mixing the functional feed and the basic fish feed according to the mass ratio of 3:7 to obtain the feed; the basic fish feed consists of the following components in percentage by mass: 70% of corn flour, 10% of fish meal, 15% of bean cake powder and 5% of bran.
Comparative example 1
A fish feed, its preparation method includes the following steps:
(1) Oven drying residue of XIASANGJU (produced by Guangzhou white cloud mountain Star (pharmaceutical industry) stock, and pulverizing and sieving with 10 mesh sieve;
(2) Fermenting the sieved mixture; the fermentation steps are as follows: inoculating Aspergillus niger and fermenting at 32deg.C for 2d; then inoculating bacillus and fermenting for 1d at 34 ℃; finally inoculating lactobacillus and yeast, fermenting at 40deg.C for 1d;
(3) After fermentation is completed, drying for 3 hours at 70 ℃;
(4) Crushing the dried mixture, and sieving the crushed mixture with a 20-mesh sieve to obtain functional feed;
(5) Uniformly mixing the functional feed with the basic fish feed according to the mass ratio of 3:7, wherein the basic fish feed comprises the following components in percentage by mass: 40% of rice bran, 38% of wheat bran, 10% of bean cake, 10% of fish meal and 2% of yeast powder.
Comparative example 2
A prebiotic composition, the method of preparation comprising the steps of:
(1) Pulverizing folium Mori, sieving with 10 mesh sieve, extracting with 70deg.C water for 3 hr, concentrating the liquid to 1/2, precipitating with 2 times of 95% ethanol for 24 hr, recovering ethanol, and collecting precipitate to obtain folium Mori polysaccharide;
(2) Collecting summer Sang Jushui ethanol precipitate produced by Guangzhou white cloud mountain star (pharmaceutical industry) and recovering ethanol, and measuring and diluting the polysaccharide concentration of the traditional Chinese medicine by sulfuric acid-phenol method to reach polysaccharide concentration of 50mg/mL; meanwhile, the concentration of the mulberry leaf polysaccharide obtained in the step (1) is measured and diluted by a sulfuric acid-phenol method, so that the concentration of the mulberry leaf polysaccharide reaches 10mg/mL, and the mulberry leaf polysaccharide and the sulfuric acid-phenol method are mixed;
(3) Performing enzymolysis treatment on the polysaccharide mixture obtained in the step (2), wherein the enzymolysis treatment comprises the following steps: firstly, performing enzymolysis for 1.5 hours at 45 ℃ by using hemicellulase, then performing enzymolysis for 0.8 hour at 50 ℃ by using pectase, and finally performing enzymolysis for 3 hours at 40 ℃ by using saccharifying enzyme, and heating for 20 minutes at 85 ℃ to inactivate enzymes;
(4) Adding pretreated HDP600 macroporous resin into the solution subjected to enzymolysis in the step (3), and carrying out vibration adsorption for 3 hours at 40 ℃ and 120r/min, wherein the dosage of the macroporous resin is 0.01g/mL; and filtering to remove macroporous resin to obtain the traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition after flavone removal.
Further, the preparation of the functional fish feed by using the prebiotic composition comprises the following steps:
(1) Oven drying residue of XIASANGJU (produced by Guangzhou white cloud mountain Star (pharmaceutical industry) stock, and pulverizing and sieving with 10 mesh sieve;
(2) Adding 5% by weight of the prebiotic composition for fermentation; the fermentation steps are as follows: inoculating Aspergillus niger and fermenting at 32deg.C for 2d; then inoculating bacillus and fermenting for 1d at 34 ℃; finally, adding 1% of traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition according to the total mass, inoculating lactobacillus and yeast, and fermenting for 1d at 40 ℃;
(3) After fermentation is completed, drying for 3 hours at 70 ℃;
(4) Crushing the dried mixture, and sieving the crushed mixture with a 20-mesh sieve to obtain functional feed;
(5) Uniformly mixing the functional feed and the basic fish feed according to the mass ratio of 3:7 to obtain the feed; the basic fish feed consists of the following components in percentage by mass: 40% of rice bran, 38% of wheat bran, 10% of bean cake, 10% of fish meal and 2% of yeast powder.
Comparative example 3
A prebiotic composition, the method of preparation comprising the steps of:
(1) Pulverizing folium Mori, sieving with 10 mesh sieve, extracting with 70deg.C water for 3 hr, concentrating the liquid to 1/2, precipitating with 2 times of 95% ethanol for 24 hr, recovering ethanol, and collecting precipitate to obtain folium Mori polysaccharide;
(2) Collecting summer Sang Jushui ethanol precipitate produced by Guangzhou white cloud mountain star (pharmaceutical industry) and recovering ethanol, and measuring and diluting the polysaccharide concentration of the traditional Chinese medicine by sulfuric acid-phenol method to reach polysaccharide concentration of 50mg/mL; meanwhile, the concentration of the mulberry leaf polysaccharide obtained in the step (1) is measured and diluted by a sulfuric acid-phenol method, so that the concentration of the mulberry leaf polysaccharide reaches 10mg/mL;
(3) Mixing the traditional Chinese medicine polysaccharide diluted in the step (2) with mulberry leaf polysaccharide, and performing ultrasonic degradation, wherein the ultrasonic power is 300W, and the degradation time is 10min; the purpose of sonication is to reduce polysaccharide viscosity;
(4) Adding pretreated HDP600 macroporous resin into the solution subjected to ultrasonic degradation in the step (3), and carrying out vibration adsorption for 3 hours at 40 ℃ and 120r/min, wherein the dosage of the macroporous resin is 0.01g/mL; and filtering to remove macroporous resin to obtain the traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition after flavone removal.
Further, the preparation of the functional fish feed by using the prebiotic composition comprises the following steps:
(1) Oven drying residue of XIASANGJU (produced by Guangzhou white cloud mountain Star (pharmaceutical industry) stock, and pulverizing and sieving with 10 mesh sieve;
(2) Adding 5% by weight of the prebiotic composition for fermentation; the fermentation steps are as follows: inoculating Aspergillus niger and fermenting at 32deg.C for 2d; then inoculating bacillus and fermenting for 1d at 34 ℃; finally, adding 1% of traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition according to the total mass, inoculating lactobacillus and yeast, and fermenting for 1d at 40 ℃;
(3) After fermentation is completed, drying for 3 hours at 70 ℃;
(4) Crushing the dried mixture, and sieving the crushed mixture with a 20-mesh sieve to obtain functional feed;
(5) Uniformly mixing the functional feed and the basic fish feed according to the mass ratio of 3:7 to obtain the feed; the basic fish feed consists of the following components in percentage by mass: 40% of rice bran, 38% of wheat bran, 10% of bean cake, 10% of fish meal and 2% of yeast powder.
Comparative example 4
A prebiotic composition, the method of preparation comprising the steps of:
(1) Pulverizing folium Mori, sieving with 10 mesh sieve, extracting with 70deg.C water for 3 hr, concentrating the liquid to 1/2, precipitating with 2 times of 95% ethanol for 24 hr, recovering ethanol, and collecting precipitate to obtain folium Mori polysaccharide;
(2) Collecting summer Sang Jushui ethanol precipitate produced by Guangzhou white cloud mountain star (pharmaceutical industry) and recovering ethanol, and measuring and diluting the polysaccharide concentration of the traditional Chinese medicine by sulfuric acid-phenol method to reach polysaccharide concentration of 50mg/mL; meanwhile, the concentration of the mulberry leaf polysaccharide obtained in the step (1) is measured and diluted by a sulfuric acid-phenol method, so that the concentration of the mulberry leaf polysaccharide reaches 10mg/mL;
(3) Mixing the traditional Chinese medicine polysaccharide diluted in the step (2) with mulberry leaf polysaccharide, and performing ultrasonic degradation, wherein the ultrasonic power is 300W, and the degradation time is 10min; the purpose of sonication is to reduce polysaccharide viscosity;
(4) Performing enzymolysis treatment on the polysaccharide mixture subjected to ultrasonic degradation in the step (3), wherein the enzymolysis treatment comprises the following specific steps:
firstly, carrying out enzymolysis for 1.5 hours at 40 ℃ by using amylase, then carrying out enzymolysis for 1 hour at 45 ℃ by using hemicellulase, finally carrying out enzymolysis for 3 hours at 45 ℃ by using lipase, and heating for 20 minutes at 85 ℃ to inactivate enzymes;
(5) Adding pretreated HDP600 macroporous resin into the solution subjected to enzymolysis in the step (4), and carrying out vibration adsorption for 3 hours at 40 ℃ and 120r/min, wherein the dosage of the macroporous resin is 0.01g/mL; and filtering to remove macroporous resin to obtain the traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition after flavone removal.
Further, the preparation of the functional fish feed by using the prebiotic composition comprises the following steps:
(1) Oven drying residue of XIASANGJU (produced by Guangzhou white cloud mountain Star (pharmaceutical industry) stock, and pulverizing and sieving with 10 mesh sieve;
(2) Adding 5% by weight of the prebiotic composition for fermentation; the fermentation steps are as follows: inoculating Aspergillus niger and fermenting at 32deg.C for 2d; then inoculating bacillus and fermenting for 1d at 34 ℃; finally, adding 1% of traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition according to the total mass, inoculating lactobacillus and yeast, and fermenting for 1d at 40 ℃;
(3) After fermentation is completed, drying for 3 hours at 70 ℃;
(4) Crushing the dried mixture, and sieving the crushed mixture with a 20-mesh sieve to obtain functional feed;
(5) Uniformly mixing the functional feed and the basic fish feed according to the mass ratio of 3:7 to obtain the feed; the basic fish feed consists of the following components in percentage by mass: 40% of rice bran, 38% of wheat bran, 10% of bean cake, 10% of fish meal and 2% of yeast powder.
Comparative example 5
A prebiotic composition, the method of preparation comprising the steps of:
(1) Pulverizing folium Mori, sieving with 10 mesh sieve, extracting with 70deg.C water for 3 hr, concentrating the liquid to 1/2, precipitating with 2 times of 95% ethanol for 24 hr, recovering ethanol, and collecting precipitate to obtain folium Mori polysaccharide;
(2) Collecting summer Sang Jushui ethanol precipitate produced by Guangzhou white cloud mountain star (pharmaceutical industry) and recovering ethanol, and measuring and diluting the polysaccharide concentration of the traditional Chinese medicine by sulfuric acid-phenol method to reach polysaccharide concentration of 50mg/mL; meanwhile, the concentration of the mulberry leaf polysaccharide obtained in the step (1) is measured and diluted by a sulfuric acid-phenol method, so that the concentration of the mulberry leaf polysaccharide reaches 10mg/mL;
(3) Mixing the traditional Chinese medicine polysaccharide diluted in the step (2) with mulberry leaf polysaccharide, and performing ultrasonic degradation, wherein the ultrasonic power is 300W, and the degradation time is 10min; the purpose of sonication is to reduce polysaccharide viscosity;
(4) Performing enzymolysis treatment on the polysaccharide mixture subjected to ultrasonic degradation in the step (3), wherein the enzymolysis treatment comprises the following specific steps: firstly, performing enzymolysis for 1.5 hours at 45 ℃ by using hemicellulase, then performing enzymolysis for 0.8 hour at 50 ℃ by using pectase, and finally performing enzymolysis for 3 hours at 40 ℃ by using saccharifying enzyme, and heating for 20 minutes at 85 ℃ to inactivate enzymes;
(5) Adding pretreated HDP600 macroporous resin into the solution subjected to enzymolysis in the step (4), and carrying out vibration adsorption for 3 hours at 40 ℃ and 120r/min, wherein the dosage of the macroporous resin is 0.01g/mL; and filtering to remove macroporous resin to obtain the traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition after flavone removal.
Further, the preparation of the functional fish feed by using the prebiotic composition comprises the following steps:
(1) Oven drying residue of XIASANGJU (produced by Guangzhou white cloud mountain Star (pharmaceutical industry) stock, and pulverizing and sieving with 10 mesh sieve;
(2) Adding 5% by weight of the prebiotic composition for fermentation; the fermentation steps are as follows: inoculating Aspergillus niger and fermenting at 32deg.C for 2d; then inoculating bacillus and fermenting for 1d at 34 ℃; finally inoculating lactobacillus and yeast, fermenting at 40deg.C for 1d;
(3) After fermentation is completed, drying for 3 hours at 70 ℃;
(4) Crushing the dried mixture, and sieving the crushed mixture with a 20-mesh sieve to obtain functional feed;
(5) Uniformly mixing the functional feed and the basic fish feed according to the mass ratio of 3:7 to obtain the feed; the basic fish feed consists of the following components in percentage by mass: 40% of rice bran, 38% of wheat bran, 10% of bean cake, 10% of fish meal and 2% of yeast powder.
Comparative example 6
A prebiotic composition, the method of preparation comprising the steps of:
(1) Pulverizing folium Mori, sieving with 10 mesh sieve, extracting with 70deg.C water for 3 hr, concentrating the liquid to 1/2, precipitating with 2 times of 95% ethanol for 24 hr, recovering ethanol, and collecting precipitate to obtain folium Mori polysaccharide;
(2) Collecting summer Sang Jushui ethanol precipitate produced by Guangzhou white cloud mountain star (pharmaceutical industry) and recovering ethanol, and measuring and diluting the polysaccharide concentration of the traditional Chinese medicine by sulfuric acid-phenol method to reach polysaccharide concentration of 50mg/mL; meanwhile, the concentration of the mulberry leaf polysaccharide obtained in the step (1) is measured and diluted by a sulfuric acid-phenol method, so that the concentration of the mulberry leaf polysaccharide reaches 10mg/mL;
(3) Mixing the traditional Chinese medicine polysaccharide diluted in the step (2) with mulberry leaf polysaccharide, and performing ultrasonic degradation, wherein the ultrasonic power is 300W, and the degradation time is 10min; the purpose of sonication is to reduce polysaccharide viscosity;
(4) Performing enzymolysis treatment on the polysaccharide mixture subjected to ultrasonic degradation in the step (3), wherein the enzymolysis treatment comprises the following specific steps: firstly, performing enzymolysis for 1.5 hours at 45 ℃ by using hemicellulase, then performing enzymolysis for 0.8 hour at 50 ℃ by using pectase, and finally performing enzymolysis for 3 hours at 40 ℃ by using saccharifying enzyme, and heating for 20 minutes at 85 ℃ to inactivate enzymes;
(5) Adding pretreated HDP600 macroporous resin into the solution subjected to enzymolysis in the step (4), and carrying out vibration adsorption for 3 hours at 40 ℃ and 120r/min, wherein the dosage of the macroporous resin is 0.01g/mL; and filtering to remove macroporous resin to obtain the traditional Chinese medicine oligosaccharide and mulberry leaf oligosaccharide prebiotic composition after flavone removal.
Further, the preparation of the functional fish feed by using the prebiotic composition comprises the following steps:
(1) Oven drying residue of XIASANGJU (produced by Guangzhou white cloud mountain Star (pharmaceutical industry) stock, and pulverizing and sieving with 10 mesh sieve;
(2) Adding 5% by weight of the prebiotic composition for fermentation; the fermentation steps are as follows: inoculating lactobacillus and yeast, fermenting at 40deg.C for 1d, inoculating bacillus, fermenting at 34deg.C for 1d, adding 1% of traditional Chinese medicine oligosaccharide and folium Mori oligosaccharide prebiotic composition according to total mass, inoculating Aspergillus niger, and fermenting at 32deg.C for 2d;
(3) After fermentation is completed, drying for 3 hours at 70 ℃;
(4) Crushing the dried mixture, and sieving the crushed mixture with a 20-mesh sieve to obtain functional feed;
(5) Uniformly mixing the functional feed and the basic fish feed according to the mass ratio of 3:7 to obtain the feed; the basic fish feed consists of the following components in percentage by mass: 40% of rice bran, 38% of wheat bran, 10% of bean cake, 10% of fish meal and 2% of yeast powder.
Verification example 1
The fish feeds prepared in comparative examples 1 to 6 of example 1 were used for grass carp feeding, and feed coefficient, weight gain rate and carcass rate were measured. The feed coefficient is also called as feed coefficient and meat increasing coefficient, and refers to the weight of the consumed feed increased by a unit weight of a breeding object. The specific calculation method comprises the following steps: feed coefficient = feed consumption/weight gain x 100%. The measurement results are shown in table 1 below.
TABLE 1 influence of example 1 and comparative examples 1-6 on grass carp growth index
The lower the feed coefficient is, the higher the feed conversion rate is, and the better the feed use effect is. The feed coefficient of example 1 is lower than that of each comparative example, which shows that the feed of the feeding example has obvious weight gain for grass carp growth, and the feed is less in intake and more in weight gain.
The weight gain rate is an index reflecting the work result of aquatic product feeding, and refers to the ratio of the weight gain of aquatic products to the initial weight of aquatic products in a certain period. Weight gain = (last weight-initial weight)/initial weight x 100%. The weight gain rate of example 1 is higher than that of each comparative example, which shows that the feed of the example is obviously increased for the growth rate of grass carp, and grass carp can grow rapidly within a certain time.
Carcass rate is the ratio of edible parts of fish to total fish. The carcass ratio of example 1 was higher than that of each comparative example, demonstrating that feeding grass carp with the feed of the examples increases the edible portion of the fish body.
Subsequently, the effect of the fish feeds prepared in example 1 and comparative examples 1 to 6 on the muscle quality of grass carp was examined. The texture profile analysis method is adopted to simulate the chewing motion of the human mouth to carry out texture measurement on the dorsum muscle of grass carp, so as to obtain the texture characteristic parameters such as hardness, elasticity, chewing force and the like. Stiffness and elasticity are the primary texture indicators affecting muscle quality. Generally, the meat quality of fish is hardened, the elasticity is enhanced, and the taste is better. The detection results are shown in the following table 2.
TABLE 2 influence of example 1 and comparative examples 1-6 on grass carp muscle quality
The hardness, elasticity and chewing force of example 1 are all higher than those of each comparative example, which shows that grass carp fed with the feed of example 1 has better meat quality and better taste than those of the grass carp fed with the feed of comparative example.
Further, the effects of the fish feeds prepared in example 1 and comparative examples 1 to 6 on the muscle-applied components and serum index of grass carp were examined. Serum triglyceride and cholesterol concentrations are inversely related to animal hyperlipoproteinemia, diabetes, kidney disease, fatty liver and the like, and excessive triglyceride and cholesterol contents in animal blood are not beneficial to animal health. The results of the measurements are shown in Table 3 below,
TABLE 3 effects of examples and comparative examples on muscle nutritional ingredients and serum metrics of grass carp
The fat, triglyceride and cholesterol of example 1 were lower than those of each comparative example, and the protein content was higher than that of the comparative example, indicating that grass carp fed with the feed of example 1 was healthier than that of the grass carp of the comparative example, and was beneficial to improving the health of the farmed grass carp.
The above detailed description describes the analysis method according to the present invention. It should be noted that the above description is only intended to help those skilled in the art to better understand the method and idea of the present invention, and is not intended to limit the related content. Those skilled in the art may make appropriate adjustments or modifications to the present invention without departing from the principle of the present invention, and such adjustments and modifications should also fall within the scope of the present invention.
Claims (3)
1. A prebiotic functional fish feed comprising a functional feed and a basal fish feed, the functional feed comprising a grass carp chrysanthemum slag, mulberry leaves and a prebiotic composition;
the prebiotic functional fish feed is prepared by the following preparation method:
(1) Oven drying residue of XIASANGJU and folium Mori, pulverizing, and sieving;
(2) Adding the prebiotic composition for fermentation; the fermentation specifically comprises the following steps: adding prebiotic composition 0.1-10% by weight, inoculating Aspergillus niger, fermenting at 30-35deg.C for 1-3d; then inoculating bacillus and fermenting for 1d at 30-37 ℃; finally adding 0.01-5% of prebiotic composition according to the total mass, inoculating lactobacillus and yeast, and fermenting at 37-45deg.C for 1-2d;
(3) After fermentation is completed, drying treatment is carried out;
(4) Crushing and sieving the dried mixture to obtain functional feed;
(5) Adding basic fish feed into the functional feed, and uniformly mixing to obtain the feed;
the prebiotic composition is prepared by the following method:
(a) Pulverizing folium Mori, sieving, extracting with water, concentrating, precipitating with ethanol, recovering ethanol, and collecting precipitate to obtain folium Mori polysaccharide;
(b) Collecting the water-extracted alcohol precipitate of the grass mulberry and the chrysanthemum, recovering the ethanol, and measuring and diluting the concentration of the traditional Chinese medicine polysaccharide; meanwhile, the concentration of the mulberry leaf polysaccharide obtained in the step (1) is measured and diluted;
(c) Mixing the traditional Chinese medicine polysaccharide diluted in the step (b) with mulberry leaf polysaccharide, and performing ultrasonic degradation;
(d) Performing enzymolysis treatment on the polysaccharide mixture subjected to ultrasonic degradation in the step (c); the enzymolysis treatment specifically comprises the following steps: firstly, performing enzymolysis for 1-2 hours at 30-55 ℃ by using hemicellulase, then performing enzymolysis for 0.5-1 hour at 30-60 ℃ by using pectase, finally performing enzymolysis for 2-4 hours at 30-50 ℃ by using saccharifying enzyme, and heating for 20min at 85 ℃ to inactivate enzyme;
(5) Adding pretreated macroporous resin into the solution subjected to enzymolysis in the step (4) for oscillating adsorption, and then filtering to remove the macroporous resin.
2. The prebiotic functional fish feed of claim 1, wherein the conditions of sonication in step (c) are: the ultrasonic power is 300W, and the degradation time is 2-20min.
3. The prebiotic functional fish feed of claim 1 wherein the functional feed and basal fish feed are used in a 3:7 ratio.
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| CN112626148A (en) * | 2020-12-11 | 2021-04-09 | 广东省农业科学院蚕业与农产品加工研究所 | Preparation method of synbiotics |
| CN113143825A (en) * | 2021-05-10 | 2021-07-23 | 广东省农业科学院蚕业与农产品加工研究所 | A collutory containing Chinese medicinal prebiotics and its preparation method |
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| CN108371231A (en) * | 2018-01-30 | 2018-08-07 | 通威股份有限公司 | A kind of tilapia feed additive and preparation method thereof |
| CN112626148A (en) * | 2020-12-11 | 2021-04-09 | 广东省农业科学院蚕业与农产品加工研究所 | Preparation method of synbiotics |
| CN113143825A (en) * | 2021-05-10 | 2021-07-23 | 广东省农业科学院蚕业与农产品加工研究所 | A collutory containing Chinese medicinal prebiotics and its preparation method |
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