CN104222643A - Hericium caput-medusae fermentation extract feed additive - Google Patents
Hericium caput-medusae fermentation extract feed additive Download PDFInfo
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Abstract
The invention discloses a hericium caput-medusae fermentation extract feed additive which is an extract obtained by performing inoculation, shake-flask culture and seeding tank fermentation on hericium caput-medusae strains and performing steaming and filtration on fermenting liquid under the temperature of 95-100 DEG C and concentrating filtrate; 0.05-0.75 percent of a hericium caput-medusae fermentation extract is added into a feed, so that the daily gain of meat chicken can be increased, and growth of animals is obviously promoted; the daily gain is increased by 8.96 percent; the feed conversion rate is increased, and the productivity of the animals is improved; meanwhile, the quality of poultry meat can be also improved, and the cholesterol content of chicken meat is reduced; the contents of cholesterols in breast muscles, leg muscles and livers of the meat chickens are respectively reduced by 30.67 percent, 48.18 percent and 57.04 percent.
Description
Technical field
The invention belongs to feed additive field, specifically Hericium caput-medusae fermentation medicinal extract feed addictive and the feed for the production of low cholesterol animal products.
Background technology
Antibacterial, the antiviral large problem being puzzlement always and cultivating industry, and use the residue problem of antibiotics to cause countries in the world to pay much attention to.From 1 day January in 2006, European Union member countries completely forbade foodstuff animal and use antibiotic feed additive for promoting growth.So find a kind of green, efficient, safe feed addictive seems very urgent.In recent years, application beneficial microbe carries out the development and production of feed addictive, for feed addictive provides a new development approach, for food industry, feed industry and its feeding provide a kind of healthy, nontoxic, free of contamination selection.
Small acupuncture Hericium erinaceus (Bull. Ex Fr.) Pers [
hericium caput-medusae(Bull.:Fr.) Pers.] be a kind of rare edible and medicinal fungi.Modern pharmacology research shows, Small acupuncture Hericium erinaceus (Bull. Ex Fr.) Pers has raising body's hypoxia tolerance and painstaking effort output quantity, accelerates blood circulation, and remove interior free yl, improve immunity, the effects such as inhibition tumor cell growth, especially have good protective effect to gastric mucosa.Artificial fermentation's medicinal extract that scientific research personnel finds to utilize Small acupuncture Hericium erinaceus (Bull. Ex Fr.) Pers to ferment and obtains has very large similitude with Hericium caput-medusae fructification in active ingredient, pharmacologically active and clinical effect etc.Hericium caput-medusae fermentation medicinal extract is except containing except polysaccharide, also have the materials such as amino acid, trace element, vitamin, phenols, terpene, there is pharmacologically active widely, now use it for widely in the treatment of the disease of digestive systems such as treatment indigestion, gastric ulcer, duodenal ulcer, cancer of the esophagus, cancer of the stomach, duodenal cancer and tumour.The existing multiple marketing drugs deriving from Hericium caput-medusae fermentation medicinal extract at present.
At present the research of Hericium caput-medusae is all only conceived to the exploitation of medicine or health food, at home and abroad on market, there is not yet the Related product utilizing Small acupuncture Hericium erinaceus (Bull. Ex Fr.) Pers to make feed addictive.
Summary of the invention
The pure natural fodder additive that the object of the invention is to provide a kind of growth promotion, provides immunocompetence, reduces foodstuff animal cholesterol level, Hericium caput-medusae fermentation medicinal extract feed addictive.
Hericium caput-medusae fermentation medicinal extract feed addictive, it is that Hericium caput-medusae bacterial strain is through inoculation, Shaking culture, seeding tank fermentation; Fermentation liquor 95-100 DEG C of boiling, filters, the medicinal extract that filtrate is condensed into;
The culture medium of described seeding tank fermentation is: 0.3% glucose, 0.03%KH
2pO
4, 0.3% peptone, 0.05% sodium chloride, Cobastab
1, defoamer, surplus is water;
Described medicinal extract polyoses content is not less than 5%.
For the production of a feed for low cholesterol animal products, it is in feed, with the addition of the above-mentioned Hericium caput-medusae fermentation medicinal extract feed addictive of 0.05-0.75%;
Described addition is 0.3%.
The invention provides Hericium caput-medusae fermentation medicinal extract feed addictive, it is that Hericium caput-medusae bacterial strain is through inoculation, Shaking culture, seeding tank fermentation; Fermentation liquor 95-100 DEG C of boiling, filters, the medicinal extract that filtrate is condensed into; The Hericium caput-medusae fermentation medicinal extract that 0.05-0.75% is above-mentioned is with the addition of in feed, broiler chicken daily gain can be improved, remarkable promotion growth of animal, daily gain improves 8.96%, improve food conversion ratio, improve breeding performonce fo animals, the quality of poultry can also be promoted simultaneously, reduce chicken cholesterol level, in chicken chest, leg flesh, liver, the content of cholesterol reduces by 30.67%, 48.18% and 57.04% respectively.
Detailed description of the invention
the preparation of embodiment 1 Hericium caput-medusae fermentation medicinal extract
(1) slant strains is cultivated:
In desinfection chamber super-clean bench rapidly by pure for Hericium caput-medusae inoculation in Special slope culture medium, cultivate 7-9 days at 26-28 DEG C, obtain liquid shaking bottle bacterial classification; Described Special slope culture medium prescription is: potato 100: glucose 1: potassium dihydrogen phosphate 1: agar 1, pH nature;
Method for making: 1000 ml that add water of fresh potato 200 g peeling being cut into small pieces boil 25 minutes, filtrate is got by filtered through gauze, add after agar soaks in advance in filtrate to boil and constantly stir simultaneously, until dissolve completely, then add the material such as glucose, potassium dihydrogen phosphate respectively to stir, be sub-packed in while hot in test tube, be stoppered bottle stopper and be placed on autoclave sterilizing, steam pressure 1.1 atmospheric pressure, 120 DEG C, 30 minutes time, after taking out, Temperature fall is to 40-45 DEG C, be paved into inclined-plane, blank cultivation 1 day, can use without miscellaneous bacteria.
(2) Shaking culture:
One-level shaking flask is inoculated: in desinfection chamber super-clean bench by tampon beyond the Great Wall after the liquid shaking bottle bacterial classification access shaking flask that obtains in described step (1), put bottle swingging machine and cultivate;
Second-level shake flask is inoculated: take off rapidly I and II shaking flask bottle stopper in desinfection chamber super-clean bench, poured into rapidly in second-level shake flask by first order seed, beyond the Great Wall tampon, cover gauze, fasten cotton rope, puts bottle swingging machine and cultivates.
Condition of culture: I and II shaking flask is placed in bottle swingging machine and turns in 180-230, shaken cultivation 4-7 days under 26-28 DEG C of condition, 4 DEG C of refrigerators are for subsequent use, obtain the liquid seeds that can be used as first class seed pot;
Described shake-flask seed culture medium prescription (I and II shaking flask method for making is identical)
Formula: 3-4% glucose, 0.03%KH
2pO
4, 0.3% peptone, 0.05% sodium chloride, appropriate vitamin B
1, a little defoamer.
Method for making: mentioned reagent is used distilled water heating for dissolving, interior loading amount 100 ml of 500 ml bottles (one-level shaking flask), 2000 ml bottle (second-level shake flask) interior loading amount 500 ml, be stoppered tampon, load autoclave in an atmospheric pressure, 121 DEG C of sterilizings 30 minutes, be cooled to room temperature and place one day, for subsequent use;
(3) first class seed pot fermentation: the condensed water in deduction sterilization process, then adds raw material and the defoamer of fluid nutrient medium, pass into vapours and be heated to 120 DEG C in seeding tank by formula, under a 0.12-0.13 MPa atmospheric pressure, 30-40 minute; Liquid seeds access step (2) obtained after sterilizing cooling is cultivated, inoculum concentration 5-10%, tank temperature 26-28 DEG C, throughput 1:0.5(V/Vmin), tank pressure 0.05 MPa, through 5-7 days ventilate cavity, bacterial classification reached exponential phase, obtains seed culture fluid.Liquid Culture based formulas used is: 0.3% glucose, 0.03%KH
2pO
4, 0.3% peptone, 0.05% sodium chloride, appropriate vitamin B
1, a little defoamer, all the other are water.
(4) ferment tank: the condensed water in deduction sterilization process, then adds fermentation culture medium raw material and defoamer by formula, pass into vapours and be heated to 120 DEG C in fermentation tank, under a 0.2 MPa atmospheric pressure, 30 minutes; After sterilizing cooling, the seed liquor access fermentation tank in first class seed pot, step (3) obtained, inoculum concentration 5-10%, ventilate cavity, tank temperature 28 DEG C, speed of agitator 150r/m, throughput 1:0.4-0.5(V/Vmin), tank pressure 0.05 MPa, incubation time 5-7 days, obtains zymotic fluid.Fermentation culture medium prescription used is: 0.3% glucose, 0.03%KH
2pO
4, 0.3% peptone, 0.05% sodium chloride, appropriate vitamin B
1, a little defoamer, all the other are water.
(5) zymotic fluid is heated: logical steam is heated to fermentation tank, keeps 10 minutes after temperature is added to 100 DEG C.
(6) filtrate is filtered: zymotic fluid is crossed plate-frame filtering, and can not leak material, filtrate asks for a clarification transparent, and filtrate flows into storage in material-storage jar.Chemical examination polyoses content is not less than 5%.
Fehlings reagent is adopted to the mensuration of total reducing sugar in zymotic fluid: get zymotic fluid 1ml and be placed in 25ml colorimetric cylinder, add 2mol/L H
2sO
415ml, puts into boiling water molten 15 minutes colorimetric cylinder; The each 5ml of film first, second test solution is added in 250ml triangular flask; Intermediate after boil-off is cooled rapidly, inserts in 100ml volumetric flask after adjusting pH6.8 with 40%NaOH, repeatedly rinse colorimetric cylinder constant volume; The intermediate 5ml getting pH6.8, in the 250ml triangular flask got ready, puts 1200W electric furnace and makes it seethe with excitement, timing 1 minute from boiling; With 0.1% glucose standard titration; Writing down and consuming number is y; Measure blank; Add each 5ml of film first, second liquid in 250ml triangular flask, do not add intermediate, directly with 0.1% glucose standard titration, consumption number is x, then calculates.Sum=(x-y) × 2
(7) filtrate simmer down to medicinal extract: tank temperature control between 60 ~ 80 DEG C, about steam pressure 0.04-0.06 MPa; Vacuum tank vacuum degree control is in 0.06-0.09 MPa.Reinforced when the heating tube gone up most will expose, each reinforced will adding less, spares no effort to, about 30 minutes.Be concentrated to all material when all having entered, be combined by the material of two cylinders, vacuum declined before putting cylinder, tank temperature brings up to about 90 DEG C.Material is concentrated to heating tube topmost and exposes sampling survey mother-in-law U.S. degree, then diligently will survey mother-in-law U.S. degree and control namely to want discharging at 33-38.When every sub-sampling, blowing, first termination of pumping, after vacuum back to zero, proportion is surveyed in sampling again.During discharging barrelling, polysaccharide is surveyed in sampling, and assay method is with the mensuration of total reducing sugar in zymotic fluid in (6).Expect in thorough purge tank.Namely concentrated extract obtains described Hericium caput-medusae fermentate feed addictive.
the mensuration of embodiment 1 Small acupuncture Hericium erinaceus (Bull. Ex Fr.) Pers liquid extract
This product be Hericium Hericium caput-medusae (
hericium caput-medusae(Bull.:Fr.) Pers.) zymotic fluid and the concentrated extract of mycelia extract.Polyoses content must not lower than 5%(Fehlings reagent).This product shows brown, the micro-perfume (or spice) of gas, mildly bitter flavor.
Polyoses content: glucose standard curve preparation is accurate takes the DEXTROSE ANHYDROUS reference substance 0.1g that 105 DEG C are dried to constant weight, puts in 100ml volumetric flask, is dissolved in water, and be diluted to scale, shake up, and obtains (containing DEXTROSE ANHYDROUS 1mg in every 1ml).
The preparation precision of calibration curve measure Glucose standards solution 0.2,0.4,0.6,0.8,1.0,1.2,1.4,1.6ml, be placed in 25ml measuring bottle respectively, add water to 2.0ml, precision adds 3 respectively, 5-dinitrosalicylic acid test solution 1.5ml, mixing, heat 5 minutes in boiling water bath, take out, be cooled to room temperature with cold water immediately, be diluted with water to scale, shake up, use water as blank, measure absorbance according to AAS (Pharmacopoeia of the People's Republic of China version in 2005 two annex IV A) at the wavelength place of 520nm, drawing standard curve.
This product 1g is got in the preparation of need testing solution, accurately weighed, puts in 50ml measuring bottle, and add water 30ml, 50 DEG C of heating water baths 20 minutes, takes out, is cooled to room temperature, is diluted with water to scale, shake up, and filters, to obtain final product.
The mensuration precision of determination method total reducing sugar measures need testing solution 2ml and puts in 25ml measuring bottle, adds 6mol/L hydrochloric acid solution 5ml, water 5ml, heat 30 minutes in boiling water bath, add instructions phenolphthalein solution 1 after cooling, be adjusted to blush with 20% sodium hydroxide solution, add water to scale, filter, precision measures filtrate 1ml, according to the method under calibration curve preparation, rise to " adding water to 2.0ml ", measure absorbance in accordance with the law, from calibration curve, read the concentration value of sugar, to obtain final product.
The mensuration precision of monose measures need testing solution 2ml, puts in 10ml measuring bottle, adds water to scale, shake up, filter, accurate absorption filtrate 1ml, according to the method under calibration curve preparation, rises to " adding water to 2.0ml ", measure absorbance in accordance with the law, from calibration curve, read the concentration value of sugar, to obtain final product.
The concentration of polysaccharide presses the concentration of total reducing sugar, deducts the concentration of monose.
The main physical and chemical index of Hericium caput-medusae fermentation extract product have selected the large nutrient of feed six and effective active composition, measures as table 1.
embodiment 3 Hericium caput-medusae fermentation medicinal extract promotes growth of meat chicken test
1, materials and methods
1.1 experimental animals and grouping
Select 1 age in days Arbor Acres (AA) broiler chicken 3000, male and female ratio half and half, body weight (36.25 scholar 0.38).Without exception through observing 6h, then it is divided into 2 groups at random, every component 3 repetition, each repetition is 500.
1.2 experimental animal daily ration composition and nutrition
Test feed raw material corn, Soybean Meal, calcium monohydrogen phosphate meet the requirement of state health standards (GB/13078-2001).With reference to national meat fowl feeding standard (NY/T33-1986) and NRC(1994) home poultry raising standard design daily ration, Hericium caput-medusae fermentation medicinal extract according to 0.3% and 0%(control group) ratio to add in basal diet (not adding any medicated premix in daily ration) and to stir and evenly mix.
1.3 broiler feeding experiment date and management
All test chickens are all raised in same hen house, are divided into 6 communities with plastic wire.Be responsible for by special messenger, ground is flat supports, brood time every square metre 30, finishing period (4-8 age in week) 12 every square metre.Use charging basket artificial feeding, free choice feeding.Freely drink water with water fountain.First week room temperature of brooding controls at 31 DEG C-32 DEG C, reduces 2-3 DEG C weekly later, remains on 23-25 DEG C after surrounding.24h illumination in first week, later natural lighting.Regularly carry out the immunity inoculation of vaccine, experimental period is 42d.
2 Hericium caput-medusaes fermentation medicinal extract are on the impact of the full phase gaining effect of chick and feed efficiency
Concrete result of the test is in table 2:
Can find out that in feed, to add the Hericium caput-medusae medicinal extract that ferments feeds AA broiler chicken by result in table 2, add 0.3% and can effectively improve its gaining effect, daily gain and the blank group difference of test group reach the level of signifiance (P<0.05), improve 8.96% than control group daily gain.Add the conversion ratio that 0.3% culture significantly can improve feed in addition in daily ration, difference reaches the level of signifiance (P<0.05) compared with control group.
embodiment 4 Hericium caput-medusae fermentation medicinal extract is on the impact of broiler chicken immunologic function
1, material
Test broiler chicken is 14,21,28 age in days AA commercial meat birds.To add the Broiler chicks group of hello Hericium caput-medusae fermentation medicinal extract for test group, addition is 0.3%; The broiler chicken group of not adding the nursing of Hericium caput-medusae fermentation medicinal extract in daily ration is blank group.At random extract each 10 plumages of donor stem cell infusion from each group, butcher peeling through bloodletting, get its thymus gland respectively, spleen, the bursa of farbricius weigh, Computation immunity shoot formation.
Immune Organs Index=immune organ quality/body weight × 100
2, data process&analysis
Experimental data adopts SPSS 19.0 statistical software to carry out statistical analysis.
3, results and analysis
The analysis result of thymus index, index and spleen index, bursal index is in table 3:
As shown in Table 3, when 14 age in days, test group broiler chicken thymus index is significantly higher than blank group (P<0.05); When 21 age in days, test group index and spleen index is significantly higher than blank group (P<0.05); The bursal index of each test group is all higher than blank group, though without significant difference (P>0.05), but each test group its bursa of farbricius catagen speed compared with blank group slows down, illustrate that Hericium caput-medusae fermentation medicinal extract delays degeneration to it and plays certain effect, Hericium caput-medusae fermentation medicinal extract is by improving the weight of immune organ and then carrying out enhanced machine body immunity function.
example 5 Hericium caput-medusae fermentation medicinal extract is on the impact of cholesterol level in chicken
1, material
Test broiler chicken is 42 age in days AA commercial meat birds.To add the Broiler chicks group of hello Hericium caput-medusae fermentation medicinal extract for test group, addition is 0.3%; The broiler chicken group of not adding the nursing of Hericium caput-medusae fermentation medicinal extract in daily ration is blank group.Extract each 10 plumages of donor stem cell infusion from each group at random, butcher peeling through bloodletting, get liver and the fresh chest muscle of same area, leg muscle immediately, frozen to be analyzed.
2, detection method
2.1 cholesterol level
The sample chest muscle collected, leg flesh, liver are taken out, shred with scissors as far as possible, be placed on plate and stir, accurately take 1g and add the homogenate of homogenate medium, method according to describing in National Standard of the People's Republic of China GB/T9695.24-2008 " meat quail Determination of Cholesterol Content " carries out chest muscle, leg flesh, hepatic cholesterol assay, because the lipid in meat quail is after saponification, cholesterol is extracted as unsaponifable matter, with gas chromatography determination, quantified by external standard method.
2.2 data process&analysis
Experimental data adopts SPSS 19.0 statistical software to carry out statistical analysis.
3, results and analysis
The analysis result of chest muscle, leg flesh, hepatic cholesterol content is in table 4:
As shown in Table 4, the Hericium caput-medusae fermentation medicinal extract of interpolation 0.3% in daily ration significantly can reduce the content (P<0,05) of cholesterol in chicken chest, leg flesh, liver, and content reduces by 30.67%, 48.18% and 57.04% respectively.
the different adding proportion of embodiment 6 Hericium caput-medusae fermentation medicinal extract is on the impact of growth of meat chicken
1, materials and methods
1.1 experimental animals and grouping
Select 1 aa broiler chicken 3000, male and female ratio half and half, body weight (35.72 scholar 0.41).Without exception through observing 6h, then it is divided into 6 groups at random, often organizes 500.
1.2 experimental animal daily ration composition and nutrition
Test feed raw material corn, Soybean Meal, calcium monohydrogen phosphate meet the requirement of state health standards (GB/13078-2001).With reference to national meat fowl feeding standard (NY/T33-1986) and NRC(1994) home poultry raising standard design daily ration, Hericium caput-medusae fermentation medicinal extract is according to 0%(control group), 0.05%, 0.1%, 0.25%,, the ratio of 0.5% and 0.75% to be added in basal diet (not adding any medicated premix in daily ration) and is stirred and evenly mixed.
13 broiler feeding experiment date and management
All test chickens are all raised in same hen house, are divided into 6 communities with plastic wire.Be responsible for by special messenger, ground is flat supports, brood time every square metre 30, finishing period (4-8 age in week) 12 every square metre.Use charging basket artificial feeding, free choice feeding.Freely drink water with water fountain.First week room temperature of brooding controls at 31 DEG C-32 DEG C, reduces 2-3 DEG C weekly later, remains on 23-25 DEG C after surrounding.24h illumination in first week, later natural lighting.Regularly carry out the immunity inoculation of vaccine, experimental period is 42d.
2, the different adding proportion of Hericium caput-medusae fermentation medicinal extract is to the shadow of the full phase gaining effect of chick and feed efficiency
Ring, concrete result of the test is in table 5:
The different adding proportion of table 5 Hericium caput-medusae fermentation medicinal extract is to chick growth effect
As can be seen from Table 5, the Hericium caput-medusae fermentation medicinal extract adding 0.1-0.5% in basal feed can significantly improve daily gain and the feed conversion rate of chick, promotes the growth of chick.And adding proportion is higher than 0.5%, the daily gain of chick can not be improved further.
Claims (5)
1. Hericium caput-medusae fermentation medicinal extract feed addictive, it is that Hericium caput-medusae bacterial strain is through inoculation, Shaking culture, seeding tank fermentation; Fermentation liquor 95-100 DEG C of boiling, filters, the medicinal extract that filtrate is condensed into.
2. Hericium caput-medusae fermentation medicinal extract feed addictive according to claim 1, is characterized in that: the culture medium of described seeding tank fermentation is: 0.3% glucose, 0.03%KH
2pO
4, 0.3% peptone, 0.05% sodium chloride, Cobastab
1, defoamer, surplus is water.
3. Hericium caput-medusae fermentation medicinal extract feed addictive according to claim 1, is characterized in that: described medicinal extract polyoses content is not less than 5%.
4., for the production of a feed for low cholesterol animal products, it is in feed, with the addition of the above-mentioned Hericium caput-medusae fermentation medicinal extract feed addictive of 0.05-0.75%.
5. a Ju a kind of feed for the production of low cholesterol animal products according to claim 4, is characterized in that: described addition is 0.3%.
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| CN104886358A (en) * | 2015-03-27 | 2015-09-09 | 吉林农业大学 | Ganoderma applanatum fermented extract paste feed additive |
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