CN105192309A - Composite feed additive - Google Patents

Abstract

The invention relates to a composite feed additive and a preparation method thereof, and belongs to the field of feed enzymic preparations. The composite feed additive is prepared from, by weight, 10-15 parts of antimicrobial peptides, 10-20 parts of propolis, 15-25 parts of glossy privet fruit extract, 15-30 parts of rhodotorula culture, 25-50 parts of toadstool enzymolysis powder, 5-10 parts of scutellaria baicalensis extract and 0.01-0.02 part of tea polyphenol, wherein rhodotorula is rhodotorula benthica WYN1 (Rhodotorula sp. WYN1), and the preservation number is CCTCC No: M 2014592. Due to the fact that the rhodotorula used in the method can produce carotene with a high yield, and inorganic copper in a culture medium is converted into chelated copper to be enriched, growth and development of animals are better promoted, the utilization rate of feed is increased, and the heat stability of the feed additive is improved.

Description

A kind of composite feed additive

Technical field:

The invention belongs to field of fodder, particularly relate to feed addictive.

Background technology:

In modern livestock and poultry cultivation process, in order to prevention and therapy Animal diseases, the excessive antibiotic product of normal use guarantees animal health, but excessive antibiotic use often causes the quality of animal meat product and product thereof to decline and antibiotic resistance strengthens, and causes human health to be also subject to having a strong impact on of antibiotic problem by the transmission of food chain.

Be derived from more natural natural materials and not only there is good nutritive peculiarity, also there is the effects such as antiviral, anti-inflammatory, anti-oxidant, conditioner body immunity function simultaneously, have broad application prospects.

The main component of propolis is flavone compound, phenols, lactone, Coumarins, aldehyde ketone, the trace element such as a small amount of iron, calcium, silicon, manganese, lead, nickel, aluminium, copper, zinc and vitamin B complex, vitamin A, and several amino acids, enzyme, polysaccharide and abundant arsanilic acid etc." natural antibiotics " that propolis or both at home and abroad expert generally acknowledge, because it not only has the effect of the external invader of defence and infection pathogen diffusion, also has the effects such as antiviral, anti-inflammatory, anti-oxidant, conditioner body immunity function.

The fruit of glossy privet is also known as glossy privet reality, purpleflower holly fruit, Chinese wax tree, mouse Chinese catalpa.Fruit of glossy privet fruit is containing oleanolic acid (Oleanolicacid), sweet mellow wine, glucose, palmitic acid, stearic acid, oleic acid and leukotrienes.Pericarp is containing ursolic acid (Ursolicacid), oleanolic acid, acetyloleanolic acid (Acetyloleanolicacid).Seed fatty oily 14.9%, in oil, palmitic acid and stearic acid are 19.5%, oleic acid and leukotrienes etc. is 80.5%.The still trace element such as cupric, zinc, iron, manganese in the fruit of glossy privet.

Fruit of glossy privet tonifying liver kidney yin, crow must improving eyesight, and modern medicine study thinks that the fruit of glossy privet can suppress the effect of helicobacter pylori to treat stomach trouble, also has the treatment suppressing purine abnormal metabolism for gout and hyperuricemia.For the deficiency of liver-yin and kidney-yin, soreness of waist tinnitus, poliosis; Unseeing, poor vision; Fever due to yin deficiency, stomach trouble and gout and and hyperuricemia.The fruit of glossy privet is clinical conventional Chinese herbal medicine simply, has begun one's study it in recent years as the effect of feed addictive in Production of Livestock and Poultry.

Antibacterial peptide is the biological small peptides that a class has antibacterial activity, is the important component part of natural animal immune defense system.The stable in physicochemical property of antibacterial peptide, relative molecular mass is little, has a broad antifungal spectrum, and antibacterial mechanisms is unique.Compared with conventional antibiotic, have antibacterial activity strong, have no drug resistance, noresidue and the little advantage of toxicity.In recent years, antibacterial peptide is the focus of research as antibiotic ideal substitute always.It is reported, antibacterial peptide, as additive feeding animals, has and improves immune performance, the generation of minimizing disease and the effect of raising survival rate.Meanwhile, antibacterial peptide has good growth promoting function to animal, can improve the growth performance of animal, reduces feedstuff-meat ratio.Therefore, antibacterial peptide has the potentiality becoming green safety feed addictive.

Baikal skullcap root is the dry root of the labiate root of large-flowered skullcap (ScutellariabaicalensisGeorgi), is a kind of conventional Chinese herbal medicine.Flavone compound is the principle active component of the root of large-flowered skullcap, and the content of baicalin (Baicalin) is the highest.Inside and outside bacteriostatic experiment shows, skullcapflavone all has bacteriostasis to Escherichia coli, staphylococcus aureus, Pseudomonas aeruginosa.

In addition, in field of animal feed, mainly with the source of the Inorganic Coppers such as copper sulphate as copper, but it is low to there is absorptivity in Inorganic Copper, there is the problems such as antagonism with other nutriments, chelated copper then can well solve the problem, it can alleviate the antagonism with other nutriments, promote the utilization of copper and other mineral matter elements, promote growing of animal, improve efficiency of feed utilization, improve carcass quality, improve reproductive performance, also there is antibiooxidation and a series of function such as immunity and good stability simultaneously, preferably economic benefit can be brought.

Carotenoid is a class in Polyenes that is yellow, orange or red, and it is the most ubiquity of occurring in nature is also the most stable natural colouring matter, is extensively present in many animals and plants.Carotenoid as a kind of excellent additive and the nutritional supplement improving nutrition, its fine quality and effect, for a long time just generally acknowledge by countries in the world, be described as most promising antioxidant.At present, carotenoid is widely used in feed additive industry, the application prospect good due to it and the function of brilliance, for a long time extremely people's favor.It is significant to feed additive industry development how effective exploitation obtains green feed additive product with natural plants and fungi.

Summary of the invention:

The invention provides a kind of composite feed additive;

Composite feed additive weight fraction is composed as follows:

Antibacterial peptide 10-15 part, propolis 10-20 part, fruit of glossy privet extract 15-25 part, rhodotorula culture 15-30 part, hickory chick enzymolysis powder 25-50 part, Baical Skullcap root P.E 5-10 part, Tea Polyphenols 0.01-0.02 part;

Rhodotorula used in the present invention, be the rich copper ocean rhodotorula WYN1 of a strain High Yield of Carotenoid, described bacterial strain on November 25th, 2014 be preserved in China typical culture collection center (address: China. Wuhan. Wuhan University, postcode 430072), deposit number is: CCTCCNo:M2014592, Classification And Nomenclature: ocean rhodotorula WYN1 (Rhodotorulasp.WYN1).

Described antibacterial peptide, propolis and Tea Polyphenols are commercially available prod;

Described fruit of glossy privet extract preparation method is as follows:

The fruit of glossy privet is pulverized, and after crossing 40-50 mesh sieve, add fruit of glossy privet 3-6 times of weight absolute ethyl alcohol Soakage extraction, adjusting temperature after control temperature 30-45 DEG C, 2-4 hour is 55-60 DEG C of 1-2 hour, and extract is concentrated, drying obtains ethanol extract; Add 75-85 DEG C of hot water in fruit of glossy privet residue after alcohol extract, hot water addition is 2-4 times of fruit of glossy privet residue weight, and processing time 30-50 minute, extracts 2-3 time continuously, by spraying dry after extract Vacuum Concentration, obtain hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, crosses 60 mesh sieves, obtain glossy privet fruit extract.

The preparation method of described hickory chick enzymolysis powder is as follows:

(1) Morchella esculenta (L.) Pers sporophore drying and crushing;

(2) water-soluble homogeneous: the Morchella esculenta (L.) Pers sporophore after pulverizing is added in stainless steel cylinder, add fructification quality 3-6 water doubly, soak 3-5 hour, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: adjustment colloid mill stator and the gap of rotor are 0.5-1 micron, colloid mill flow be 0.4-1 ton/hour;

(3) intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 50-60 DEG C, adjustment pH to 4.5-6.0, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05-0.1%, the 1,4 beta-glucanase of 0.01-0.1%, the protease of 0.01-0.1%, insulation enzymolysis 0.5-1.5 hour, constantly stirs in enzymolysis process;

(4) dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis.

The preparation method of described Baical Skullcap root P.E is as follows:

The root of large-flowered skullcap is put in the supersonic wave cleaning machine filling 0.1-0.3% sodium bicarbonate solution and clean 10-15min in 200W, 30KHz, drain, being crushed to particle diameter is below 2mm, put Homogeneous phase mixing in container and add the water of 3-6 times of weight, control temperature 40-50 DEG C keeps 2-4h, then being warming up to 70 DEG C-90 DEG C, is 3.5-4.5 by lactic acid adjust ph, under power 150-300W condition, carry out Microwave Extraction 10-15min; Then the complex enzyme adding mixed material gross weight 0.5-1.5% carries out enzymolysis, be 5.5-6.8 by lactic acid adjust ph, adjust the temperature to 40-50 DEG C, enzymolysis 2-4h, finally add the mixture of mixed material 1-3 times of w ethanol and propyl alcohol, the mass ratio of ethanol and propyl alcohol mixing is 1:1-3, control temperature to 50 DEG C-60 DEG C of maintenance 3-4h, filter, obtain the first filtrate; Add the water of filter residue 1-3 times of weight, control temperature 65 DEG C-75 DEG C keeps 1-3h, is then cooled to 25-35 DEG C, filters, obtains the second filtrate; First filtrate and the second filtrate are merged according to mass ratio 2-4:1-3, namely Homogeneous phase mixing obtains root of large-flowered skullcap extract; Namely filtrate obtain Baical Skullcap root P.E through ultrafiltration, reduced pressure concentration, freeze drying.

Described complex enzyme is dextranase, zytase, pentosanase, pectase 3-5:2-4:1-3:1-3 Homogeneous phase mixing in mass ratio.

Described rhodotorula culture preparation method is as follows:

Seed culture is cultivated by yeast conventional culture methods;

Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 3-8%, and shaking speed is 50-80r/min, cultivate for 24-28 DEG C and to adjust temperature after 10-15 hour and be 16-20 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 3-5, keeps 2-4 hour; Stairstepping is warmed up to 24-30 DEG C afterwards, and pH is adjusted to 4.6-5.5, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 50-100r/min, continues fermentation 10-15h.

After fermentation ends, 10-1000 μm, fermentation liquor aperture coarse filtration, then concentratedly with 10-20 DEG C of loop ultrafiltration removes moisture and obtains solid content 20-40% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding.

Fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 1-3%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 150-200mg/L, sodium chloride 20%, and all the other are water;

The preparation method of described enzymolysis corn flour is: by corn flour material-compound tank, be that 2:1 adds pure water with V:m, adjust ph 3-7, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, and warming while stirring is to 45-55 DEG C of insulation 15-30min, then be slowly warming up to 60-65 DEG C of insulation 15-30min, freeze drying is for subsequent use.

Described composite feed additive is obtained by following methods:

After pulverizing, hickory chick enzymolysis powder, antibacterial peptide, propolis, fruit of glossy privet extract, Baical Skullcap root P.E, Tea Polyphenols and rhodotorula culture are proportionally mixed to get composite feed additive.

Beneficial effect:

1, the present invention utilizes the rich copper ocean rhodotorula WYN1 being separated from marine environment and obtaining a plant height product carrotene, effectively raises carrotene and copper output in yeast by static gas wave refrigerator method.Adopt said method fermented and cultured ocean rhodotorula WYN1, in its biomass, carotenoid output and born of the same parents, copper content is respectively more than 27g/L, the dry bacterium of 35-40mg/L and 7240-8000 μ g/g.In research and development, inventor surprisingly finds that low temperature static gas wave refrigerator effectively can improve carrot and copper content, is important achievement of the present invention.

2, because rhodotorula used in the present invention can high yield carrotene, and the Inorganic Copper in culture medium is converted into chelated copper and carries out enrichment, better facilitate growing of animal, improve efficiency of feed utilization, can 58.6% be improved for milking sow weight of weaning litter, piglet head daily gain improves 41.7%, diarrhea rate reduces 54.1%, improves 48.91% for weanling pig daily gain, and feedstuff-meat ratio improves 14.1%, diarrhea rate reduces 10%, and death rate improves 3.7%.

3, because chelated copper has good heat endurance, feed addictive provided by the present invention is had and preserves active and heat endurance preferably, make the preservation activity of additive improve 35%, heat endurance improves more than 25%

4, the fruit of glossy privet of the present invention carries out pulverize and break cellular wall process, function factor in the fruit of glossy privet is made to realize full price stripping and high efficiency extraction utilization, the effective extraction, particularly control temperature stage extraction that adopt organic solvent and hot water to realize wherein different efficacies composition respectively effectively improve extraction efficiency and the quality of glossy privet fruit extract especially.

5, the present invention is composite by science, rhodotorula, hickory chick are achieved organic assembling, particularly add the traditional Chinese medicine ingredients such as the root of large-flowered skullcap, product of the present invention effectively can reduce the use amount of antibiotics in letting animals feed, improve the security of animal meat product, improve the food utilization efficiency of animal, strengthen the immunity of animal, improve raise benefit.

Detailed description of the invention:

Embodiment 1: the acquisition of ocean rhodotorula bacterium WYN1 and the domestication of resistance to copper ability

(1) by gathering the fresh biological specimen such as extra large shrimp, starfish, marine alga from Huanghai Sea seashore, be placed in aseptic YEPD culture medium, picking redness or pink circular colonies, be further purified cultivation.According to the accumulation of each bacterial classification carotenoid, the bacterial classification that screening carotenoid output is high.The ocean rhodotorula WYN1 that wherein carotenoid output is higher, cell is oval, does not have pseudohypha; The YEPD culture medium of seawater configuration forms red colonies, smooth surface, neat in edge; Product spore culture medium does not produce ascospore, and polygon budding, without ballistopore; Azymic sugar, does not form kind of starch compound, does not assimilate inositol.According to These characteristics, tentatively determine to belong to Rhodotorula (Rhodotorula).Inclined-plane is stored in refrigerator.

(2) fermented and cultured of ocean rhodotorula WYN1

Activate on strain transfer to slant medium, after 28 DEG C of cultivation 24h, be inoculated in liquid seed culture medium, 28 DEG C of shaken cultivation 36h obtain seed liquor, be equipped with in the 250mL triangular flask of 60mL fermentation medium by 10% inoculum concentration access, 28 DEG C, shaken cultivation 48h (shaking speed is 180r/min).Fermentation medium consists of glucose 2%, peptone 1%, yeast extract 1%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, salinity 20.48h cultivated by 28 DEG C of shaking tables, and rotating speed is 180r/min.

(3) resistance to copper ability domestication

Ocean rhodotorula bacterium WYN1 being inoculated in content of copper ion is in 50mg/L (copper sulphate configuration) culture medium, is placed in 28 DEG C, and on 180r/min constant-temperature table, 24h is cultivated in concussion.Then from this culture, get 3ml fermentate is inoculated in the malt extract medium of not cupric, cultivates 24h.Repeatedly cultivate several generations, yeast is grown in the culture medium of cupric stable, and when quantity no longer increases, more progressively improve the copper concentration of culture medium.So saccharomycete repeatedly just can be made can to adapt to the environment that content of copper ion is 300mg/l, obtain the characteristic of resistance to copper.The present invention, after ocean rhodotorula being carried out to the domestication of resistance to copper ability, can improve cellular biomass, the output of carotenoid and the rich copper ability of yeast effectively.

(4) mensuration of cellular biomass

By centrifugal for zymotic fluid 3000r/min 10min, abandon supernatant, thalline is again centrifugal after aseptic washing 2 ~ 3 times, and gained thalline is placed in 60 DEG C of baking ovens and is dried to constant weight, correct amount.

(5) extraction of carotenoid:

1g dry mycelium is put into 100mL triangular flask, and add the hydrochloric acid soaking at room temperature 1h of 5mL3mol/L, boiling water bath 4min, cools rapidly, and 3000r/min is centrifugal, and 15min is precipitated as cell residue, is settled to 20mL extraction, obtains carotenoid leaching liquor with acetone.

(6) mensuration of carotenoid

After suitably being diluted by extract, under 475nm condition, measure absorbance value with 722 type spectrophotometers.Be calculated as follows carotenoid content:

In formula: A λ max is the absorbance at 475nm wavelength place; D is extension rate when measuring sample; V is acetone consumption (mL); 0.16 is the molar extinction coefficient of carotenoid; W is rhodotorula dry cell weight (g).

(7) yeast copper content measures

Use aas determination.

(8) culture presevation

The above-mentioned rich copper ocean rhodotorula WYN1 through domestication is preserved in China typical culture collection center, and deposit number is: CCTCCNo:M2014592.

In carotenoid production and born of the same parents, chelated copper content is respectively 27.52g/L, 25.12mg/L and the dry bacterium of 6240 μ g/g.

Embodiment 2 one kinds of composite feed additives

Composite feed additive weight fraction is composed as follows:

Antibacterial peptide 12 parts, propolis 15 parts, fruit of glossy privet extract 20 parts, rhodotorula culture 20 parts, 40 parts, hickory chick enzymolysis powder, Baical Skullcap root P.E 8 parts, Tea Polyphenols 0.015 part; Rhodotorula is CCTCCNo:M2014592;

Antibacterial peptide is Chu Tai bio tech ltd, Shanghai sell goods;

Propolis, Tea Polyphenols are commercially available;

Described fruit of glossy privet extract preparation method is as follows:

The fruit of glossy privet is pulverized, and crosses after 45 mesh sieves, adds the fruit of glossy privet 5 times of weight absolute ethyl alcohol Soakage extraction, control temperature 40 DEG C, adjusts temperature and be 58 DEG C and maintain 1.5 hours after 3 hours, and concentrated, the drying of extract afterwards obtains ethanol extract; Add 80 DEG C of hot water in fruit of glossy privet residue after alcohol extract, hot water addition is 3 times of fruit of glossy privet residue weight, 40 minutes processing times, extracts 2 times continuously, by spraying dry after extract Vacuum Concentration, obtains hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, crosses 60 mesh sieves, obtain glossy privet fruit extract.

The preparation method of described hickory chick enzymolysis powder is as follows:

(1) Morchella esculenta (L.) Pers sporophore drying and crushing is to particle diameter 1mm;

(2) water-soluble homogeneous: the Morchella esculenta (L.) Pers sporophore after pulverizing is added in stainless steel cylinder, add the water of fructification quality 5 times, soak 4 hours, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: the gap of adjustment colloid mill stator and rotor is 0.8 micron, and colloid mill flow is 0.8 ton/hour;

(3) intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 55 DEG C, pH is to 5.0 in adjustment, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.08%, the 1,4 beta-glucanase of 0.05%, the protease of 0.05%, insulation enzymolysis 1 hour, constantly stirs in enzymolysis process;

(4) dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis.

The preparation method of described Baical Skullcap root P.E is as follows:

The root of large-flowered skullcap is put in the supersonic wave cleaning machine filling 0.2% sodium bicarbonate solution and clean 12min in 200W, 30KHz, drain, being crushed to particle diameter is 2mm, put Homogeneous phase mixing in container and add the water of 5 times of weight, control temperature 45 DEG C keeps 3h, then being warming up to 80 DEG C, is 4.0 by lactic acid adjust ph, under power 200W condition, carry out Microwave Extraction 12min; Then the complex enzyme adding mixed material gross weight 1% carries out enzymolysis, be 6 by lactic acid adjust ph, adjust the temperature to 45 DEG C, enzymolysis 3h, finally add the mixture of mixed material 2 times of w ethanol and propyl alcohol, the mass ratio of ethanol and propyl alcohol mixing is 1:2, and control temperature to 55 DEG C keeps 3.5h, filter, obtain the first filtrate; Add the water of filter residue 2 times of weight, control temperature 70 DEG C keeps 2h, is then cooled to 30 DEG C, filters, obtains the second filtrate; First filtrate and the second filtrate are merged according to mass ratio 3:2, namely Homogeneous phase mixing obtains root of large-flowered skullcap extract; Namely filtrate obtain Baical Skullcap root P.E through ultrafiltration, reduced pressure concentration, freeze drying.

Described complex enzyme is dextranase, zytase, pentosanase, pectase 4:3:2:2 Homogeneous phase mixing in mass ratio.

Described rhodotorula culture preparation method is as follows:

Rhodozyma culture method carries out seed culture routinely;

Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 5%, and shaking speed is 65r/min, cultivate for 26 DEG C and to adjust temperature after 12 hours and be 18 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 4, keeps 3 hours; Do not have a hour intensification 2 DEG C to 26 DEG C afterwards, pH is adjusted to 5, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 80r/min, continues fermentation 12h;

After fermentation ends, 500 μm, fermentation liquor aperture coarse filtration, then concentratedly with 15 DEG C of loop ultrafiltrations removes moisture and obtains solid content 30% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding.

Fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 2%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 180mg/L, sodium chloride 20%, and all the other are water;

The preparation method of described enzymolysis corn flour is: by corn flour material-compound tank, be that 2:1 adds pure water with V:m, adjust ph 5, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, warming while stirring to 50 DEG C insulation 25min, then be slowly warming up to 62 DEG C of insulation 25min, freeze drying is for subsequent use.

Adopt said method fermented and cultured ocean rhodotorula WYN1, in zymotic fluid, in biomass, carotenoid output and born of the same parents, chelated copper content is respectively 32.52g/L, 40mg/L and the dry bacterium of 8000 μ g/g.

Described composite feed additive is obtained by following methods:

After pulverizing, hickory chick enzymolysis powder, antibacterial peptide, propolis, fruit of glossy privet extract, Baical Skullcap root P.E, Tea Polyphenols and rhodotorula culture are proportionally mixed to get composite feed additive.

Embodiment 3 one kinds of composite feed additives

Composite feed additive parts by weight are composed as follows:

Antibacterial peptide 10 parts, propolis 10 parts, fruit of glossy privet extract 15 parts, rhodotorula culture 15 parts, 50 parts, hickory chick enzymolysis powder, Baical Skullcap root P.E 5 parts, Tea Polyphenols 0.01 part; Rhodotorula is CCTCCNo:M2014592;

Antibacterial peptide is Chu Tai bio tech ltd, Shanghai sell goods;

Propolis, Tea Polyphenols are commercially available;

Described fruit of glossy privet extract preparation method is as follows:

The fruit of glossy privet is pulverized, and crosses after 40 mesh sieves, adds the fruit of glossy privet 3 times of weight absolute ethyl alcohol Soakage extraction, control temperature 30 DEG C, adjusts temperature and be 55 DEG C and maintain 1 hour after 2 hours, and concentrated, the drying of extract afterwards obtains ethanol extract; Add 75 DEG C of hot water in fruit of glossy privet residue after alcohol extract, hot water addition is 2 times of fruit of glossy privet residue weight, 30 minutes processing times, extracts 2 times continuously, by spraying dry after extract Vacuum Concentration, obtains hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, crosses 60 mesh sieves, obtain glossy privet fruit extract.

The preparation method of described hickory chick enzymolysis powder is as follows:

(1) Morchella esculenta (L.) Pers sporophore drying and crushing is to particle diameter 2mm;

(2) water-soluble homogeneous: the Morchella esculenta (L.) Pers sporophore after pulverizing is added in stainless steel cylinder, add the water of fructification quality 3 times, soak 3 hours, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: the gap of adjustment colloid mill stator and rotor is 0.5 micron, and colloid mill flow is 0.4 ton/hour;

(3) intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 50 DEG C, pH is to 4.5 in adjustment, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05%, the 1,4 beta-glucanase of 0.01%, the protease of 0.01%, insulation enzymolysis 0.5 hour, constantly stirs in enzymolysis process;

(4) dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis.

The preparation method of described Baical Skullcap root P.E is as follows:

The root of large-flowered skullcap is put in the supersonic wave cleaning machine filling 0.1% sodium bicarbonate solution and clean 10min in 200W, 30KHz, drain, being crushed to particle diameter is 1mm, put Homogeneous phase mixing in container and add the water of 3 times of weight, control temperature 40 DEG C keeps 2h, then being warming up to 70 DEG C DEG C, is 3.5 by lactic acid adjust ph, under power 150W condition, carry out Microwave Extraction 10min; Then the complex enzyme adding mixed material gross weight 0.5% carries out enzymolysis, be 5.5 by lactic acid adjust ph, adjust the temperature to 40 DEG C, enzymolysis 2h, finally add the mixture of mixed material 1 times of w ethanol and propyl alcohol, the mass ratio of ethanol and propyl alcohol mixing is 1:1, control temperature to 50 DEG C DEG C maintenance 3h, filter, obtain the first filtrate; Add the water of filter residue 1 times of weight, control temperature 65 DEG C DEG C keeps 1h, is then cooled to 25 DEG C, filters, obtains the second filtrate; First filtrate and the second filtrate are merged according to mass ratio 2:1, namely Homogeneous phase mixing obtains root of large-flowered skullcap extract; Namely filtrate obtain Baical Skullcap root P.E through ultrafiltration, reduced pressure concentration, freeze drying.

Described complex enzyme is dextranase, zytase, pentosanase, pectase 3:2:1:1 Homogeneous phase mixing in mass ratio.

Described rhodotorula culture preparation method is as follows:

Seed culture is cultivated by yeast conventional culture methods;

Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 3%, and shaking speed is 50r/min, cultivate for 24 DEG C and to adjust temperature after 10 hours and be 16 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 3, keeps 2 hours; Intensification per hour 1.5 DEG C is afterwards warmed up to 24-30 DEG C, and pH is adjusted to 4.6, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 50r/min, continues fermentation 10h.

After fermentation ends, 100 μm, fermentation liquor aperture coarse filtration, then concentratedly with 10 DEG C of loop ultrafiltrations removes moisture and obtains solid content 20% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding.

Fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 2%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 150mg/L, sodium chloride 20%, and all the other are water;

The preparation method of described enzymolysis corn flour is: by corn flour material-compound tank, be that 2:1 adds pure water with V:m, adjust ph 3, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, warming while stirring to 45 DEG C insulation 15min, then be slowly warming up to 60 DEG C of insulation 15min, freeze drying is for subsequent use.

Adopt said method fermented and cultured ocean rhodotorula WYN1, in zymotic fluid, in biomass, carotenoid output and born of the same parents, chelated copper content is respectively 29.38g/L, 35mg/L and the dry bacterium of 7240 μ g/g.

Described composite feed additive is obtained by following methods:

After pulverizing, hickory chick enzymolysis powder, antibacterial peptide, propolis, fruit of glossy privet extract, Baical Skullcap root P.E, Tea Polyphenols and rhodotorula culture are proportionally mixed to get composite feed additive.

Embodiment 4 one kinds of composite feed additives

Composite feed additive parts by weight are composed as follows:

Antibacterial peptide 15 parts, propolis 20 parts, fruit of glossy privet extract 25 parts, rhodotorula culture 30 parts, 25 parts, hickory chick enzymolysis powder, Baical Skullcap root P.E 10 parts, Tea Polyphenols 0.02 part; Rhodotorula is CCTCCNo:M2014592;

Antibacterial peptide is Chu Tai bio tech ltd, Shanghai sell goods;

Propolis, Tea Polyphenols are commercially available;

Described fruit of glossy privet extract preparation method is as follows:

The fruit of glossy privet is pulverized, and after crossing 50 mesh sieves, adds the fruit of glossy privet 6 times of weight absolute ethyl alcohol Soakage extraction, control temperature 45 DEG C, and adjusting temperature after 4 hours is 60 DEG C of maintenances 2 hours, and extract is concentrated, drying obtains ethanol extract; Add 85 DEG C of hot water in fruit of glossy privet residue after alcohol extract, hot water addition is 4 times of fruit of glossy privet residue weight, 50 minutes processing times, extracts 3 times continuously, by spraying dry after extract Vacuum Concentration, obtains hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, crosses 60 mesh sieves, obtain glossy privet fruit extract.

The preparation method of described hickory chick enzymolysis powder is as follows:

(1) Morchella esculenta (L.) Pers sporophore drying and crushing is to particle diameter 2mm;

(2) water-soluble homogeneous: the Morchella esculenta (L.) Pers sporophore after pulverizing is added in stainless steel cylinder, add the water of fructification quality 6 times, soak 5 hours, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: the gap of adjustment colloid mill stator and rotor is 1 micron, and colloid mill flow is 1 ton/hour;

(3) intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 60 DEG C, pH is to 6.0 in adjustment, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.1%, the 1,4 beta-glucanase of 0.1%, the protease of 0.1%, insulation enzymolysis 1.5 hours, constantly stirs in enzymolysis process;

(4) dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis.

The preparation method of described Baical Skullcap root P.E is as follows:

The root of large-flowered skullcap is put in the supersonic wave cleaning machine filling 0.3% sodium bicarbonate solution and clean 15min in 200W, 30KHz, drain, being crushed to particle diameter is below 2mm, put Homogeneous phase mixing in container and add the water of 6 times of weight, control temperature 50 DEG C keeps 4h, then being warming up to 90 DEG C, is 4.5 by lactic acid adjust ph, under power 300W condition, carry out Microwave Extraction 15min; Then the complex enzyme adding mixed material gross weight 1.5% carries out enzymolysis, be 6.8 by lactic acid adjust ph, adjust the temperature to 50 DEG C, enzymolysis 4h, finally add the mixture of mixed material 3 times of w ethanol and propyl alcohol, the mass ratio of ethanol and propyl alcohol mixing is 1:3, and control temperature to 60 DEG C keeps 4h, filter, obtain the first filtrate; Add the water of filter residue 3 times of weight, control temperature 75 DEG C keeps 3h, is then cooled to 35 DEG C, filters, obtains the second filtrate; First filtrate and the second filtrate are merged according to mass ratio 4:3, namely Homogeneous phase mixing obtains root of large-flowered skullcap extract; Namely filtrate obtain Baical Skullcap root P.E through ultrafiltration, reduced pressure concentration, freeze drying.

Described complex enzyme is dextranase, zytase, pentosanase, pectase 5:4:3:3 Homogeneous phase mixing in mass ratio.

Described rhodotorula culture preparation method is as follows:

Seed culture is cultivated by yeast conventional culture methods;

Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 8%, and shaking speed is 80r/min, cultivate for 28 DEG C and to adjust temperature after 15 hours and be 20 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 5, keeps 4 hours; 3 DEG C per hour are warmed up to 30 DEG C afterwards, and pH is adjusted to 5.5, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 100r/min, continues fermentation 15h.

After fermentation ends, 1000 μm, fermentation liquor aperture coarse filtration, then concentratedly with 20 DEG C of loop ultrafiltrations removes moisture and obtains solid content 40% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding.

Fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 3%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 200mg/L, sodium chloride 20%, and all the other are water;

The preparation method of described enzymolysis corn flour is: by corn flour material-compound tank, be that 2:1 adds pure water with V:m, adjust ph 7, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, warming while stirring to 55 DEG C insulation 30min, then be slowly warming up to 65 DEG C of insulation 30min, freeze drying is for subsequent use.

Adopt said method fermented and cultured ocean rhodotorula WYN1, in zymotic fluid, in biomass, carotenoid output and born of the same parents, chelated copper content is respectively 28.76g/L, 38mg/L and the dry bacterium of 7580 μ g/g.

Described composite feed additive is obtained by following methods:

After pulverizing, hickory chick enzymolysis powder, antibacterial peptide, propolis, fruit of glossy privet extract, Baical Skullcap root P.E, Tea Polyphenols and rhodotorula culture are proportionally mixed to get composite feed additive.

The result of use test of embodiment 5 embodiment 2 gained feed addictive in weanling pig

Test method:

Choose 120 weanling pigs, be divided into experimental group and control group by body weight, blood lineage and no sex difference, often organize 3 repetitions, each repetition 20 pigs.Experiment periods is 28 days, and experimental group feed per ton adds additive 120g described in the present embodiment 2, and control group does not add additive, weighs on an empty stomach, calculate feed conversion rate, daily gain, feedstuff-meat ratio, diarrhea rate and death rate morning day at the whole story of experimental period.Experimental result is as shown in the table:

	Daily gain	Feedstuff-meat ratio	Diarrhea rate	Death rate
					Control group	226.31g	1.49	15％	5％
Experimental group	337.16g	1.28	5％	1.7％

From experimental result, the feed being added with additive of the present invention can make weanling pig daily gain improve 48.91%, and feedstuff-meat ratio improves 14.1%, and diarrhea rate reduces 10%, and death rate improves 3.7%

The result of use test of embodiment 6 example 2 of the present invention feed addictive in milking sow

Test adopts single factor test contrast design, random selecting 30 healthy, farrowing head number is close with birth counterpoise, parity is the milking sow of 2 or 3 tires, be divided at random 2 groups (i.e. test group and control groups), often organize 15 repetitions, carry out the feeding experiment of 21 days by a definite date.Wherein: test group daily ration adds the additive be prepared into by embodiment 2, and control group does not add.Record weaned piglet weight, diarrhea rate, the death rate.

Project	Test group	Control group
			Number born alive (head number)	12	10.9
Birth counterpoise (kg)	1.51	1.46
			21 days small weaning pigs (head)	10	8.30
21 days weight of weaning litters (kg)	68.5	43.2
			Wean counterpoise (kg) in 21 days	6.85	5.20
Head net gain (kg)	5.34	3.74
			Head daily gain (g)	252.28	178.1
Grice diarrhoea rate (%)	6.1	13.3

Test shows: test group can improve 58.6% than control group weight of weaning litter, and piglet head daily gain improves 41.7%, and diarrhea rate reduces 54.1%.

The above results shows that product of the present invention can improve sow and piglet body immunity, reduces antibiotic dosage, increases cultivation quality and benefits.

Embodiment 7 rhodotorula preserves active impact to additive

Rhodotorula in embodiment 2, for sample 1, is replaced with commercially available fodder yeast by the additive obtained with the preparation method described in embodiment 2, and the preservation of comparing two kinds of samples is active.At room temperature preserve above-mentioned sample 6 months, repeat the experiment of embodiment 8 afterwards, characterized the change of additive activity by the change of feedstuff-meat ratio, result shows: the residual activity of sample 1 is more than 85%, and the residual activity of sample 2 remains on about 60%.

Embodiment 8 rhodotorula is on the impact of additive heat endurance

Rhodotorula in embodiment 2, for sample 1, is replaced with commercially available fodder yeast, compares the heat endurance of two kinds of samples by the additive obtained with the preparation method described in embodiment 2.At 45 DEG C, preserve above-mentioned sample 24h, repeat the experiment of embodiment 8 afterwards, characterized the change of additive activity by the change of feedstuff-meat ratio, result shows: the residual activity of sample 1 is more than 75%, and the residual activity of sample 2 remains on about 40%.

Claims (7)

1. a composite feed additive, it is characterized in that, be made up of the component of following weight fraction: antibacterial peptide 10-15 part, propolis 10-20 part, fruit of glossy privet extract 15-25 part, rhodotorula culture 15-30 part, hickory chick enzymolysis powder 25-50 part, Baical Skullcap root P.E 5-10 part, Tea Polyphenols 0.01-0.02 part;

Described rhodotorula is specially ocean rhodotorula WYN1 (Rhodotorulasp.WYN1), and deposit number is: CCTCCNo:M2014592.

2. a kind of composite feed additive as claimed in claim 1, is characterized in that, described rhodotorula culture preparation method is as follows:

Seed culture is cultivated by yeast conventional culture methods;

Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 3-8%, and shaking speed is 50-80r/min, cultivate for 24-28 DEG C and to adjust temperature after 10-15 hour and be 16-20 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 3-5, keeps 2-4 hour; Stairstepping is warmed up to 24-30 DEG C afterwards, and pH is adjusted to 4.6-5.5, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 50-100r/min, continues fermentation 10-15h.

After fermentation ends, 10-1000 μm, fermentation liquor aperture coarse filtration, then concentratedly with 10-20 DEG C of loop ultrafiltration removes moisture and obtains solid content 20-40% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding.

3. a kind of composite feed additive as claimed in claim 2, it is characterized in that, described fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 1-3%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 150-200mg/L, sodium chloride 20%, all the other are water.

4. a kind of composite feed additive as claimed in claim 2, it is characterized in that, the preparation method of described enzymolysis corn flour is: corn flour is placed in material-compound tank, is that 2:1 adds pure water, adjust ph 3-7 with V:m, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, and warming while stirring is to 45-55 DEG C of insulation 15-30min, then be slowly warming up to 60-65 DEG C of insulation 15-30min, freeze drying is for subsequent use.

5. prepare a method for composite feed additive as claimed in claim 1, comprise the steps:

(1) described antibacterial peptide, propolis and Tea Polyphenols are commercially available prod;

(2) described fruit of glossy privet extract preparation method is as follows:

The fruit of glossy privet is pulverized, and after crossing 40-50 mesh sieve, add fruit of glossy privet 3-6 times of weight absolute ethyl alcohol Soakage extraction, adjusting temperature after control temperature 30-45 DEG C, 2-4 hour is 55-60 DEG C of 1-2 hour, and extract is concentrated, drying obtains ethanol extract; Add 75-85 DEG C of hot water in fruit of glossy privet residue after alcohol extract, hot water addition is 2-4 times of fruit of glossy privet residue weight, and processing time 30-50 minute, extracts 2-3 time continuously, by spraying dry after extract Vacuum Concentration, obtain hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, crosses 60 mesh sieves, obtain glossy privet fruit extract;

(3) preparation method of described hickory chick enzymolysis powder is as follows:

Morchella esculenta (L.) Pers sporophore drying and crushing;

Water-soluble homogeneous: the Morchella esculenta (L.) Pers sporophore after pulverizing is added in stainless steel cylinder, add fructification quality 3-6 water doubly, soak 3-5 hour, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: adjustment colloid mill stator and the gap of rotor are 0.5-1 micron, colloid mill flow be 0.4-1 ton/hour;

Intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 50-60 DEG C, adjustment pH to 4.5-6.0, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05-0.1%, the 1,4 beta-glucanase of 0.01-0.1%, the protease of 0.01-0.1%, insulation enzymolysis 0.5-1.5 hour, constantly stirs in enzymolysis process;

Dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis;

(4) preparation method of described Baical Skullcap root P.E is as follows:

The root of large-flowered skullcap is put in the supersonic wave cleaning machine filling 0.1-0.3% sodium bicarbonate solution and clean 10-15min in 200W, 30KHz, drain, being crushed to particle diameter is below 2mm, put Homogeneous phase mixing in container and add the water of 3-6 times of weight, control temperature 40-50 DEG C keeps 2-4h, then being warming up to 70 DEG C-90 DEG C, is 3.5-4.5 by lactic acid adjust ph, under power 150-300W condition, carry out Microwave Extraction 10-15min; Then the complex enzyme adding mixed material gross weight 0.5-1.5% carries out enzymolysis, be 5.5-6.8 by lactic acid adjust ph, adjust the temperature to 40-50 DEG C, enzymolysis 2-4h, finally add the mixture of mixed material 1-3 times of w ethanol and propyl alcohol, the mass ratio of ethanol and propyl alcohol mixing is 1:1-3, control temperature to 50 DEG C-60 DEG C of maintenance 3-4h, filter, obtain the first filtrate; Add the water of filter residue 1-3 times of weight, control temperature 65 DEG C-75 DEG C keeps 1-3h, is then cooled to 25-35 DEG C, filters, obtains the second filtrate; First filtrate and the second filtrate are merged according to mass ratio 2-4:1-3, namely Homogeneous phase mixing obtains root of large-flowered skullcap extract; Namely filtrate obtain Baical Skullcap root P.E through ultrafiltration, reduced pressure concentration, freeze drying;

Described complex enzyme is dextranase, zytase, pentosanase, pectase 3-5:2-4:1-3:1-3 Homogeneous phase mixing in mass ratio;

(5) described rhodotorula culture preparation method is as follows:

Seed culture is cultivated by yeast conventional culture methods;

Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 3-8%, and shaking speed is 50-80r/min, cultivate for 24-28 DEG C and to adjust temperature after 10-15 hour and be 16-20 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 3-5, keeps 2-4 hour; Stairstepping is warmed up to 24-30 DEG C afterwards, and pH is adjusted to 4.6-5.5, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 50-100r/min, continues fermentation 10-15h.

After fermentation ends, 10-1000 μm, fermentation liquor aperture coarse filtration, then concentratedly with 10-20 DEG C of loop ultrafiltration removes moisture and obtains solid content 20-40% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding;

Fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 1-3%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 150-200mg/L, sodium chloride 20%, and all the other are water;

The preparation method of described enzymolysis corn flour is: corn flour is placed in material-compound tank, be that 2:1 adds pure water with V:m, adjust ph 3-7, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, and warming while stirring is to 45-55 DEG C of insulation 15-30min, then be slowly warming up to 60-65 DEG C of insulation 15-30min, freeze drying is for subsequent use;

(6) described composite feed additive is obtained by following methods:

Hickory chick enzymolysis powder, antibacterial peptide, propolis, fruit of glossy privet extract, Baical Skullcap root P.E and rhodotorula culture are proportionally mixed to get composite feed additive.

6. a composite feed additive according to claim 1, is characterized in that, is made up of the component of following weight fraction: antibacterial peptide 12 parts, propolis 15 parts, fruit of glossy privet extract 20 parts, rhodotorula culture 20 parts, 40 parts, hickory chick enzymolysis powder, Baical Skullcap root P.E 8 parts, Tea Polyphenols 0.015 part; Rhodotorula is CCTCCNo:M2014592.

7. the application of composite feed additive according to claim 1 in animal and fowl fodder.