CN112535237A - Mangosteen fermentation extract, preparation method and application - Google Patents
Mangosteen fermentation extract, preparation method and application Download PDFInfo
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- CN112535237A CN112535237A CN202011386907.8A CN202011386907A CN112535237A CN 112535237 A CN112535237 A CN 112535237A CN 202011386907 A CN202011386907 A CN 202011386907A CN 112535237 A CN112535237 A CN 112535237A
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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
- 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
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- 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
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- 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
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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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
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- C—CHEMISTRY; METALLURGY
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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- 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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Abstract
The invention discloses a mangosteen extract with high mangostin content, and a preparation method and application thereof. The preparation method of the mangosteen extract with high mangostin content specifically comprises the following steps: pulverizing a mangosteen shell raw material to obtain mangosteen shell powder; mixing the mangosteen shell powder with water, emulsifying, stirring and cutting for multiple times to obtain mangosteen shell homogenate; uniformly mixing the mangosteen shell homogenate with the activated bacterial liquid for fermentation; performing solid-liquid separation after fermentation to obtain mangosteen shell fermentation product; mixing and extracting the mangosteen shell fermentation product with an alcohol-water extracting agent to obtain a mangosteen shell fermentation alcohol extract; concentrating and drying to obtain mangosteen extract with high mangostin content, which is suitable for being used as feed additive for resisting bacteria and promoting growth.
Description
Technical Field
The invention belongs to the field of natural plant extraction, and relates to a preparation method and application of a mangosteen extract.
Background
Antibiotics refer to a class of substances produced by bacteria, fungi, or other microorganisms that have anti-pathogenic or other activity. Due to the abuse of antibiotics in recent years, pathogenic bacteria develop resistance to antibiotics and antibiotics may remain in animal products. With the increasing emphasis on food safety in recent years, the focus is on finding antibiotics for feeding, and solving the problem of animal-derived food safety. The Chinese herbal medicine additive has unique advantages in promoting animal growth and improving immunity, so that research strength of the Chinese herbal medicine as the feed additive is further increased, and the Chinese herbal medicine anti-cancer additive has a wide application prospect in development and application of a reliable curative effect, small toxic and side effects, convenience in application and low cost.
Mangosteen, originally named mangosteen, is a plant of the genus garcinia of the family garciniaceae, and is mainly distributed in southeast Asia countries such as Thailand, Vietnam, Malaysia, Indonesia, Philippines, and the like. The pericarp of the mangosteen is always used as a traditional medicine in southeast Asia countries for treating diseases such as abdominal pain, diarrhea, dysentery, cholera, infected wounds, suppuration, chronic ulcer and the like, and modern scientific researches find that the pericarp of the mangosteen contains various active ingredients including bispyridone, phenolic acid, polysaccharide, pigment and the like, has pharmacological activities in various aspects such as anti-inflammation, antibiosis, malaria prevention, blood fat reduction, antioxidation, HIV resistance, immunoregulation and the like, is a plant resource with multiple effects, and is worthy of deep research and wide popularization. The xanthone compound is considered as a main active ingredient of mangosteen shell with drug effect and health care value, and the main active ingredient of the xanthone compound is mangostin (mangostin), so that the xanthone compound has good effect in the field of antibiosis and antiphlogosis.
At present, there are many reports related to the production process of mangosteen extract, such as chinese patent publication No. CN201310340403.6, a mangosteen extract and its application, CN111170981A, a garcinia extract extracted from mangosteen, its preparation method and its application, CN201710310190.0, a mangosteen extract for treating gout, and its preparation method. The patent methods adopt conventional extraction processes such as leaching, thermal reflux, ultrasonic and the like, and are difficult to completely break firm cell walls of the mangosteen shell powder raw material, so that effective components in plants cannot be well extracted and utilized, not only is resource waste caused, but also the prospect of the mangosteen extract as an antibacterial raw material is influenced. In fact, many polyphenols and polysaccharides exist in the mangosteen besides antibacterial substances such as mangostin, and substances with low application value in mangosteen shells are further converted into substances with high application value by a fermentation method, so that the application value of the mangosteen extract is improved. Based on the background, the invention further researches the physical and biotransformation extraction method of the mangosteen, so as to further obtain the process technology of the mangosteen extract which is more comprehensive and more suitable for being used as a feed additive for replacing antibiotics than the common extract, and particularly obtain the application method of the extract.
Disclosure of Invention
The invention aims to provide a physical-microbial composite wall-breaking extraction preparation method which is easy to operate and can efficiently produce mangosteen extract rich in mangosteen, beneficial components such as mangosteen, polyphenol, polysaccharide and the like in mangosteen, and enzymes, probiotics, polypeptides and other nutrient substances formed by fermentation are extracted at one time, and the extract is used in the field of feed additives, and is applied to the fields of antibacterial function extract, substitute antibiotics and growth promotion as feed additives.
The preparation process method related by the invention comprises the following steps: through a mechanical wet grinding mode, the mangosteen shells are gradually ground into a fine particle state in water as much as possible, then grinding liquids (homogeneous liquids) are combined, fermentation wall breaking and conversion are further carried out by a microbial fermentation method, and finally, conventional alcohol-water solution is used for extraction, and conventional spray drying is used for drying, so that the mangosteen extract with unexpected effect on antibacterial effect is obtained.
In order to achieve the purpose, the invention adopts the following process steps:
1) drying or baking fresh mangosteen, crushing the raw materials, and sieving the crushed raw materials with a 20-mesh sieve to obtain a mangosteen shell crushed material;
2) mixing the pulverized mangosteen shell with water, emulsifying, stirring and cutting in a homogenizer to obtain mangosteen shell homogenate, sterilizing and cooling; the preparation method of the mangosteen shell homogenate comprises the following steps: mixing the mangosteen shell crushed material with water according to the mass ratio of 1 (5-7), homogenizing and shearing for 0.4-1 h to obtain first fine crushed pulp and first fine crushed slag precipitated at the bottom of the water, and filtering and separating; adding water into the first fine crushed slag to carry out secondary homogenizing shearing to obtain second fine crushed pulp and second fine crushed slag deposited at the bottom of the water, and filtering and separating; homogenizing and separating for 5 times, and mixing all fine pulp to obtain mangosteen homogenate;
3) adding activated composite microbial strain culture solution into the homogenate, and performing fermentation culture at 32-36 deg.C and 160 r/min; the culture medium formula for activating the strains is 10.0g/L peptone, 10.0g/L yeast extract, 2.0g/L diammonium hydrogen citrate, 0.6 g/L magnesium sulfate, 2.0g/L dipotassium hydrogen phosphate, 0.3 g/L manganese sulfate, 1.0mL Tween-80 and the pH value is 6.2-6.6. The activated composite microbial strain is prepared by culturing lactobacillus plantarum powder and saccharomyces cerevisiae powder in a ratio of 1:3 in a liquid culture medium until the number of bacterial colonies reaches 108Liquid seed culture in CFU/ml. The inoculation mass of the composite microbial strain culture solution is 15-20% of the mass of mangosteen homogenate, and the fermentation time is 72 h;
4) filtering the fermented substance; adding 60-80% ethanol into the filter residue according to the mass ratio of 1:5-1:7, performing ultrasonic extraction for 30min, filtering, and repeating the above operation once; mixing the three filtrates, vacuum concentrating under reduced pressure to more than 10 Baume degree, stirring, homogenizing, adding appropriate adjuvants, and spray drying to obtain powdery mangosteen extract rich in mangostin, polypeptide, polyphenol, and probiotic.
The invention has the following beneficial effects:
1. the batch high shear mode of the homogenizer can break plant cell walls. The microbial fermentation further destroys the cell tissue structure of the plant by utilizing the physiological process of the microorganism, decomposes the plant tissue into small molecular substances, reduces the extraction mass transfer resistance, fully releases active ingredients, and has the characteristics of stable extraction, high purity, low energy consumption, high efficiency and the like. The physical rolling and microbial fermentation modes are combined, plant cell walls can be efficiently crushed, active ingredients are fully released, and meanwhile, carbohydrate in the mangosteen shells is used as a carbon source, so that carbohydrate and water-soluble pigment impurities are reduced, and small molecular substances which are easy to absorb by intestinal tracts and improve intestinal tract steady state are generated.
2. The invention has simple process condition, short production period and low cost, and the used raw materials are pure natural components, are safe, have no pollution, and are cheap and easy to obtain. Compared with the mangosteen extract prepared by pure water extraction and alcohol extraction, the mangosteen extract prepared by the method has the advantages that the mangosteen extract has obviously improved mangostin extraction rate, has good antibacterial rate on methicillin-resistant staphylococcus aureus and candida albicans, has obviously enhanced inhibition effect on gram negative bacteria such as escherichia coli and the like, and has wide application prospect in the field of replacing antibiotics and preservatives.
Detailed Description
The technical solutions of the present invention are further described below by specific examples, but the present invention is not limited thereto. The invention is not limited to the above embodiments, but may be modified and replaced by other embodiments.
Example 1
1) Drying 3.0 kg of fresh mangosteen, crushing the raw materials, and sieving the crushed raw materials with a 20-mesh sieve to obtain 1.17 kg of crushed mangosteen powder;
2) mixing the mangosteen shell crushed material with 6kg of water, putting the mixture into a high-speed refiner, carrying out wet grinding and stirring for 25min, and then carrying out centrifugal settling separation to obtain first fine crushed pulp and 1.40kg of first fine crushed slag (small loss is not counted) precipitated at the bottom of water; keeping the fine crushed pulp for standby, adding 6kg of water into the first fine crushed slag, carrying out second wet grinding, stirring and cutting to obtain 0.81kg of second fine crushed pulp and second settled fine crushed slag; then continuously carrying out wet grinding and stirring cutting for 3 times in a mode of slag-liquid ratio of 1:5, and finally obtaining fifth fine crushed pulp and a very small amount of fifth fine crushed slag. The five fine crushed pulps are combined to obtain 29kg of mangosteen homogenate.
3) Adding 4.35kg of activated composite microbial strain culture solution into the mangosteen homogenate, and performing fermentation culture at 32 ℃ and 160r/min for 72 h; the culture medium formula for activating the strains is 10.0g/L peptone, 10.0g/L yeast extract, 2.0g/L diammonium hydrogen citrate, 0.6 g/L magnesium sulfate, 2.0g/L dipotassium hydrogen phosphate, 0.3 g/L manganese sulfate, 1.0mL Tween-80 and the pH value is 6.2-6.6. The activated composite microbial strain is prepared by culturing lactobacillus plantarum powder and saccharomyces cerevisiae powder in a ratio of 1:3 in a liquid culture medium until the number of bacterial colonies reaches 108Liquid seed culture in CFU/ml.
4) Filtering the fermented substance to obtain 0.62 kg of filter residue; adding 0.6kg of 60% ethanol into the filter residue according to the mass ratio of 1:5, performing ultrasonic extraction for 30min, filtering to obtain 0.27kg of filter residue, and repeating the above operation once; mixing the three filtrates, vacuum concentrating under reduced pressure to 18 kg, adding dextrin 2kg, stirring, homogenizing, and spray drying to obtain 2.9 kg of mangosteen extract (with adhesive loss in spray drying stage);
and (3) taking an alpha-mangostin reference substance (with the content of 99.5%) as a standard working curve, and measuring the content of the alpha-mangostin in the obtained mangosteen extract dry powder by adopting a high performance liquid chromatography to obtain the content of the alpha-mangostin in the mangosteen extract of 5.46%.
Example 2
1) Drying 2.0kg of fresh mangosteen, crushing, and sieving with a 20-mesh sieve to obtain 768g of mangosteen shell crushed material;
2) mixing the mangosteen shell crushed material with 4kg of water, homogenizing at high speed, stirring and cutting for 30min, and performing centrifugal sedimentation to obtain 826g of first fine crushed pulp and first fine crushed slag at the bottom of precipitation; 4.5kg of water is added into the first fine crushed slag to carry out secondary high-speed homogenizing stirring and cutting to obtain second fine crushed pulp and 321g of second fine crushed slag deposited at the bottom of the water; continuously carrying out stirring separation circulation for 3 times to obtain fifth fine crushed pulp (the fifth fine crushed slag is less than 20 g); mixing the fine crushed pulp to obtain 19.3kg of mangosteen homogenate, and discarding the final fine crushed slag;
3) adding activated complex microorganism strain into the combined fine crushing paddle3.86kg of culture solution, and carrying out fermentation culture for 72h at the temperature of 32 ℃ and the rotating speed of 160 r/min; the culture medium formula for activating the strains is 10.0g/L peptone, 10.0g/L yeast extract, 2.0g/L diammonium hydrogen citrate, 0.6 g/L magnesium sulfate, 2.0g/L dipotassium hydrogen phosphate, 0.3 g/L manganese sulfate, 1.0mL Tween-80 and the pH value is 6.2-6.6. The activated composite microbial strain is prepared by culturing lactobacillus plantarum powder and saccharomyces cerevisiae powder in a ratio of 1:3 in a liquid culture medium until the number of bacterial colonies reaches 108Liquid seed culture in CFU/ml.
4) Filtering the fermented substance to obtain about 0.41kg of filter residue; adding 80% ethanol 0.7kg into the residue at a solid-to-liquid ratio of 1:7, ultrasonic treating for 30min, filtering, and repeating the above steps; mixing the three filtrates, vacuum concentrating under reduced pressure to 8kg, adding dextrin 1kg, stirring, homogenizing, and spray drying to obtain 1.4kg of mangosteen extract;
and (3) taking an alpha-mangostin reference substance (with the content of 99.5%) as a standard working curve, and measuring the content of the alpha-mangostin in the obtained mangosteen extract dry powder by adopting a high performance liquid chromatography to obtain the content of the alpha-mangostin in the mangosteen extract of 5.94%.
Example 3
The antibacterial activity of the mangosteen extract of example 1 against methicillin-resistant staphylococcus aureus, candida albicans, escherichia coli was determined: in this example, the Minimal Inhibitory Concentration (MIC) of methicillin-resistant Staphylococcus aureus, Candida albicans, and Escherichia coli activity of the sample was determined by Resazurin color development.
1) Preparing a methicillin-resistant staphylococcus aureus, candida albicans and escherichia coli liquid: respectively adopting fungus culture medium and bacteria culture medium, preparing liquid culture medium, culturing methicillin-resistant Staphylococcus aureus, Candida albicans and Escherichia coli, preparing bacteria liquid, and preparing bacterial suspension with concentration of 107-108CFU/ml；
2) Preparation of a sample solution: weighing the mangosteen extract obtained in example 1, and preparing a mangosteen extract sample solution of 2.5mg/ml (calculated by the content of alpha-mangostin) by using a dimethyl sulfoxide (DMSO) solution;
3) preparation of a resazurin developing solution: weighing 2.4mg of resazurin color developing agent, and dissolving with 24ml of deionized water;
4) preparing a resazurin-bacterium liquid mixed solution: mixing the color development liquid and the bacterial liquid according to the proportion of 3: 2;
5) 96-well plate gradient dilution assay: taking 20ul of sample solution respectively, repeating 2 holes in parallel in a first row of 96 holes, adding 180ul of the resazurin-bacterial liquid mixed solution in the first row, and adding 100ul of the resazurin-bacterial liquid mixed solution in other rows; after the first row is mixed by re-suction, 100uL of the solution is taken out and transferred to the corresponding plate hole of the second row, and the solution is multiplied and diluted to the 8 th row by the same method. Finally, the well plate added with the sample is placed into a constant temperature incubator and cultured for 8-10h at 37 ℃. The bacteria liquid turns red into no bacteriostatic activity, blue is bacteriostatic activity, and the lowest dilution concentration of the bacteria liquid maintaining blue is considered as the lowest bacteriostatic concentration of the compound to be detected. The minimum inhibitory concentration of the mangosteen extract to methicillin-resistant staphylococcus aureus and candida albicans is 18ug/ml (calculated by the content of alpha-mangostin) and the minimum inhibitory concentration of the mangosteen extract to escherichia coli is 135ug/ml (calculated by the content of alpha-mangostin) by taking vancomycin hydrochloride and kanamycin sulfate as positive controls and taking a dimethyl sulfoxide (DMSO) solution as a blank control.
Example 4
A prepared mangosteen extract with the content of alpha-mangostin of 10.2% (a control sample obtained by ultrasonic extraction twice by using 60% ethanol water solution as an extracting agent, 30min each time, concentration and spray drying) is subjected to the same dilution control test of a 96-well plate as in example 3 to compare the difference of antibacterial performance of a homogeneous fermentation extract and a common extract.
And (3) determining the Minimum Inhibitory Concentration (MIC) of the activity of methicillin-resistant staphylococcus aureus, candida albicans and escherichia coli of the sample by adopting a resazurin color development method.
1) Preparing a methicillin-resistant staphylococcus aureus, candida albicans and escherichia coli liquid: respectively adopting fungus culture medium and bacteria culture medium, preparing liquid culture medium, culturing methicillin-resistant Staphylococcus aureus, Candida albicans and Escherichia coli, preparing bacteria liquid, and preparing bacterial suspension with concentration of 107-108CFU/ml；
2) Preparation of a sample solution: weighing a conventional mangosteen extract with the content of 10.2 percent of alpha-mangostin, and preparing a mangosteen extract sample solution with the content of 2.5mg/ml (calculated by the content of the alpha-mangostin) by using a dimethyl sulfoxide (DMSO) solution;
3) preparation of a resazurin developing solution: weighing 2.4mg of resazurin color developing agent, and dissolving with 24ml of deionized water;
4) preparing a resazurin-bacterium liquid mixed solution: mixing the color development liquid and the bacterial liquid according to the proportion of 3: 2;
5) 96-well plate gradient dilution assay: taking 20ul of sample solution respectively, repeating 2 holes in parallel in a first row of 96 holes, adding 180ul of the resazurin-bacterial liquid mixed solution in the first row, and adding 100ul of the resazurin-bacterial liquid mixed solution in other rows; after the first row is re-pipetted and mixed, 100. mu.L of the solution is transferred to the corresponding well in the second row and is multiply diluted to row 8 in the same way. Finally, the well plate added with the sample is placed into a constant temperature incubator and cultured for 8-10h at 37 ℃. The bacteria liquid turns red into no bacteriostatic activity, blue is bacteriostatic activity, and the lowest dilution concentration of the bacteria liquid maintaining blue is considered as the lowest bacteriostatic concentration of the compound to be detected. By taking vancomycin hydrochloride and kanamycin sulfate as positive controls and taking dimethyl sulfoxide (DMSO) solution as a blank control, the minimum inhibitory concentration of the mangosteen extract to methicillin-resistant staphylococcus aureus and candida albicans is 27 mu g/ml (calculated by the content of alpha-mangostin), and the minimum inhibitory concentration to escherichia coli is 635ug/ml (calculated by the content of alpha-mangostin).
Therefore, under the condition of comparing solutions with the same content of alpha-mangostin, the mangosteen extract obtained by adopting the homogenization-fermentation-extraction-drying process has obviously improved inhibition effect on gram-positive bacteria and candida albicans and obviously enhanced inhibition effect on gram-negative bacteria (escherichia coli) compared with the mangosteen extract obtained by the conventional extraction method. Therefore, the mangosteen extract obtained by the invention has bacteriostasis capability superior to that of common mangosteen extracts, and has potential application prospect in the field of replacing antibiotics and antiseptics.
Example 5
The mangosteen extract obtained in example 1 was mixed with corn starch to form a premix having an α -mangostin content of 2.0% and used as a feed additive instead of antibiotics. The premix is added into daily ration of broiler chickens according to the adding proportion of 1 kilogram per ton, and the mixture is pressed into feed particles. The growth promotion experiment is carried out on 15-day-old chicks consisting of 200 experimental groups to 200 control groups for 15 days, and antibiotics and other growth promotion feed additives are not used in the experimental group and the control group in the experimental process.
The experimental results show that: the number of dead chicks in the experimental group is 2, and the weight of the chicks is increased by 2.15kg in 15 balances; the number of dead chicks in the control group is 32, and the average weight is increased by 1.78 kg.
Example 6
The mangosteen extract obtained in example 1 and corn starch are mixed into a premix with the alpha-mangostin content of 2.0%, and the premix is used as a daily ration additive for broiler chickens to replace antibiotics. The premix is added into daily ration of broiler chickens according to the adding proportion of 1.5 kg per ton, and the mixture is pressed into feed particles. The growth promotion experiment is carried out on 15-day-old chicks consisting of 200 experimental groups to 200 control groups for 15 days, and antibiotics and other growth promotion feed additives are not used in the experimental group and the control group in the experimental process.
The experimental results show that: the number of dead chicks in the experimental group is 0, and the weight of the chicks is increased by 2.20kg in 15 balances; the number of dead chicks in the control group is 28, and the average weight is increased by 1.90 kg.
Example 7
The mangosteen extract obtained in example 1 and corn starch are mixed into a premix with the alpha-mangostin content of 2.0%, and the premix is used as a daily ration additive for broiler chickens to replace antibiotics. The premix is added into daily ration of broiler chickens according to the adding proportion of 2.0kg per ton, and the mixture is pressed into feed particles. The growth promotion experiment is carried out on 15-day-old chicks consisting of 200 experimental groups to 200 control groups for 15 days, and antibiotics and other growth promotion feed additives are not used in the experimental group and the control group in the experimental process.
The experimental results show that: the number of dead chicks in the experimental group is 0, and the weight of the chicks is increased by 2.40kg in 15 balances; the number of dead chicks in the control group is 29, and the average weight is increased by 1.88 kg.
Example 8
The mangosteen extract obtained in example 1 and corn starch are mixed into a premix with the alpha-mangostin content of 2.0%, and the premix is used as a daily ration additive for broiler chickens to replace antibiotics. The premix is added into daily ration of broiler chickens according to the adding proportion of 2.0kg per ton, and the mixture is pressed into feed particles. The growth promotion experiment is carried out on 15-day-old chicks consisting of 200 experimental groups to 200 control groups for 15 days, and antibiotics and other growth promotion feed additives are not used in the experimental group and the control group in the experimental process.
The experimental results show that: the number of dead chicks in the experimental group is 0, and the weight of the chicks is increased by 2.20kg in 15 balances; the number of dead chicks in the control group is 22, and the average weight is increased by 1.80 kg.
Example 9
Commercially available mangosteen extract (17% of alpha-mangostin determined by high performance liquid chromatography) and corn starch are mixed into premix with 2.0% of alpha-mangostin, and the premix is used as broiler ration additive for replacing antibiotics. The premix is added into daily ration of broiler chickens according to the adding proportion of 2.0kg per ton, and the mixture is pressed into feed particles. The growth promotion experiment is carried out on 15-day-old chicks consisting of 200 experimental groups to 200 control groups for 15 days, and antibiotics and other growth promotion feed additives are not used in the experimental group and the control group in the experimental process.
The experimental results show that: the number of dead chicks in the experimental group is 6, and the weight of the chicks is increased by 2.08kg in 15 balances; the number of dead chicks in the control group is 25, and the average weight is increased by 1.77 kg.
Experiments show that the mangosteen extract prepared by the invention has a certain antibiotic effect on broiler chickens, has a growth promoting function, and can be used as a natural extract feed additive for replacing antibiotics to be used for poultry breeding. Although aquatic and mammalian studies have not been conducted, it is anticipated that the mangosteen extract prepared by the present invention may also have antibacterial and growth promoting effects in other animal breeding fields.
Claims (6)
1. A mangosteen fermentation extract is characterized by adopting the following preparation steps:
1) drying or baking fresh mangosteen, crushing the raw materials, and sieving the crushed raw materials with a 20-mesh sieve to obtain a mangosteen shell crushed material;
2) mixing the pulverized mangosteen shell with water, emulsifying, stirring and cutting in a homogenizer to obtain mangosteen shell homogenate, sterilizing and cooling; the preparation method of the mangosteen shell homogenate comprises the following steps: mixing the mangosteen shell crushed material with water according to the mass ratio of 1 (5-7), homogenizing and shearing for 0.4-1 h to obtain first fine crushed pulp and first fine crushed slag precipitated at the bottom of the water, and filtering and separating; adding water into the first fine crushed slag to carry out secondary homogenizing shearing to obtain second fine crushed pulp and second fine crushed slag deposited at the bottom of the water, and filtering and separating; homogenizing and separating for 5 times, and mixing all fine pulp to obtain mangosteen homogenate;
3) adding activated composite microbial strain culture solution into the homogenate, and performing fermentation culture at 32-36 deg.C and 160 r/min; the formula of the culture medium for activating the strains is 10.0g/L peptone, 10.0g/L yeast extract, 2.0g/L diammonium hydrogen citrate, 0.6 g/L magnesium sulfate, 2.0g/L dipotassium hydrogen phosphate, 0.3 g/L manganese sulfate, 1.0mL of Tween-80 and the pH value of the culture medium is 6.2-6.6;
the activated composite microbial strain is prepared by culturing lactobacillus plantarum powder and saccharomyces cerevisiae powder in a ratio of 1:3 in a liquid culture medium until the number of bacterial colonies reaches 108Liquid spawn of CFU/ml; the inoculation mass of the composite microbial strain culture solution is 15-20% of the mass of mangosteen homogenate, and the fermentation time is 72 h;
4) filtering the fermented substance; adding 60-80% ethanol into the filter residue according to the mass ratio of 1:5-1:7, performing ultrasonic extraction for 30min, filtering, and repeating the above operation once; mixing the three filtrates, vacuum concentrating under reduced pressure to more than 10 Baume degree, stirring, homogenizing, adding appropriate adjuvants, and spray drying to obtain powdery mangosteen extract rich in mangostin, polypeptide, polyphenol, and probiotic.
2. Use of the mangosteen extract of claim 1 in animal feed stocks.
3. Use of the mangosteen extract according to claim 1 in a poultry farming feed.
4. Use of the mangosteen extract of claim 1 in broiler breeding feed.
5. Use of the mangosteen extract of claim 1 in the replacement of antibiotic feed additives and feed additive raw materials.
6. Use of the mangosteen extract of claim 1 in growth-promoting feed additives and feed additive raw materials.
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| CN115590846A (en) * | 2022-10-27 | 2023-01-13 | 中国农业科学院兰州畜牧与兽药研究所（Cn） | Application of alpha-mangostin in preparation of preparation for inhibiting formation of escherichia coli biofilm |
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