CN109907168A - Pet food and preparation method thereof - Google Patents

Abstract

The invention provides a pet ration and a preparation method thereof. The pet ration uses agricultural waste residue as the main raw material, first is fermented and decomposed by the combination of black fungus, Grifola frondosa and Inonotus obliquus, then fermented by lactic acid bacteria, and finally mixed with soybean meal , the bran is mixed to prepare pet rations, the invention can effectively degrade the lignin and hemicellulose of the waste residue, improve the fermentation efficiency and the palatability of the ration, and the black fungus, frondosa and Inonotus obliquus are medicinal and edible fungi , Rich in nutrients, it can promote the pet's stomach power, promote digestion and absorption, and improve immunity. The pet ration agricultural waste residue prepared by the invention has high degradation rate, rich nutrient content, short preparation time and good development prospect.

Description

A kind of pet grain ration and preparation method thereof

Technical field

The invention belongs to feed for pet technical field, in particular to a kind of pet grain ration and preparation method thereof.

Background technique

China is large agricultural country, and agriculture waste residue such as bagasse, manioc waste are chiefly used in firing at present as one of Main By product Burning and paper pulp papermaking, with the continuous progress of science and technology, agriculture waste residue will be by more as the synthesis higher value application of high-quality resource More concerns.Main utilization ways of waste residue at present: first is that directly burning heat production power generation, accounts for 75% or so of bagasse yield；Second is that making Paper pulp papermaking raw material accounts for 20% or so of waste residue yield；Other purposes such as feed, fertilizer account for 5% or so.Agriculture waste residue it is main In ingredient, cellulose is 35% ~ 50%, hemicellulose 19% ~ 24%, lignin 23% ~ 32%, greatly affects the agreeable to the taste of feed Property, it is unfavorable for digesting.

Patent of invention CN201510360054.3, a kind of mushroom taste feed improving young companion animals dog palatability and its preparation Method, the feed include the raw material of following weight percent: mushroom powder 3.0%, dregs of beans 28.0%, peanut meal 9.0%, wheat bran 5.0%, Corn flour 35.0%, calcium monohydrogen phosphate 2.2%, live stock and fowl bone mud albumen 8.5%, edible oil 3.0%, mountain flour 2.0%, carrot meal 1.5% are made a gift of Shellfish foot-powder 2.0%, salt 0.8%；It further include 0.2% trehalose for accounting for above-mentioned raw materials gross weight, 0.08%4- hexyl resorcin, 0.02% multi-vitamins.The mushroom taste feed of invention preparation soaks in water rear feed shape and keeps complete, and palatability is good, battalion It supports and enriches comprehensively, long-term consumption can make pet dog increase immunity, promote length development and improve coat health.

Patent of invention CN201110303174.1 utilizes the method for banana lees production expanded pet feed, technical side Case is: schlempe dry, pulverize, and content of starch is added, and adjust moisture content, expanded molding.The invention can be in banana lees The minerals such as crude fibre, protein and K, Na, Ca, Mg are recycled, and are turned waste into wealth；Extrusion processing can reach simultaneously Processing and molding effect, are degraded and are modified to crude fibre, the protein in banana lees, improved it and digest and absorb Rate；Eliminate or reduce the pollution to ambient enviroment.The invention produces extrusion feed using masa parasdisiac fruit wine slag, has reached secondary Using the effect of protein, starch and dietary fiber, the nutritional ingredient of feed is improved, production technology simplifies, and the process-cycle is short, Business efficiency is high.

The research report efficiently utilized about the extraction purification of the active polysaccharide in agriculture waste residue, lignin and cellulose It is less, there is good economic benefit using agriculture waste residue preparation pet grain, but lignin and hemicellulose in agriculture waste residue It is the main reason for hindering microbial degradation cellulose that element, which is connected into firm binder course by covalent bond, is that raising grain is agreeable to the taste Property emphasis problem to be solved.

Summary of the invention

Big for agriculture waste residue degradation difficulty, the problem of the pet grain ration palatability difference of preparation, the present invention provides a kind of agriculture The method of industry produced by fermenting waste residues preparation pet grain ration.

Pet grain ration prepared by the present invention is suitable for the pets such as cat, dog, fish, hamster, bird.

The present invention is achieved by the following technical solutions:

A kind of pet grain ration, including component 1, component 2 and component 3, the component 1 is by following weight fraction ratio substance by dividing Solution bacterium and lactobacillus-fermented after be prepared: 20 ~ 40 parts of agriculture waste residue, 1 ~ 3 part of promotor, 5 ~ 10 parts of dregs of beans, 5 ~ 10 parts of wheat bran, 10 ~ 15 parts of Siraitia grosvenorii root tuber powder, 2 ~ 5 parts of salt, 2 ~ 5 parts of edible oil；The component 2 includes following weight fraction ratio substance: 2 ~ 5 parts of corn flour, 5 ~ 8 parts of wheat flour, 2 ~ 5 parts of mealy potato, 1 ~ 3 part of salt；The component 3 is by following weight fraction ratio substance Composition: 20 ~ 50 parts of meat, 5 ~ 10 parts of vitamin, 5 ~ 10 parts of minerals.

Preferably, the agriculture waste residue is one of bagasse, manioc waste.

Preferably, the promotor is one of Tween 80, oleic acid, and volume fraction is 0.1 ~ 0.5%.

Preferably, the edible oil is one of peanut oil or olive oil.

Preferably, the meat is one of pork, beef, mutton, the flesh of fish, chicken, duck or a variety of.

Preferably, the vitamin be vitamin B, it is vitamin E, vitamin B12, one or more in vitamin C.

Preferably, the minerals are one or more in K, Na, Ca, Zn, Fe.

The present invention using black fungus, Inonotus obliquus and grifola frondosus and adds organic acid and table using agriculture waste residue as main material After face activating agent promotes solid state fermentation agricultural waste residue, using lactobacillus-fermented, made of being finally mixed with dregs of beans, wheat bran Pet grain ration, the lignin and hemicellulose of the agriculture waste residue that can effectively degrade retain cellulose, improve fermentation efficiency and feeding The palatability of material, compared to liquid state fermentation, the degradation rate of lignin is much big for black fungus, Inonotus obliquus and grifola frondosus solid state fermentation In the degradation rate of cellulose, more celluloses can be retained, be more advantageous to feed nutrition demand；Black fungus, Inonotus obliquus and ash Setting flower three classes fungi is dual-purpose of drug and food bacterium, is rich in nutriment, can improve the stomach power of pet, promotes pet feed and inhales It receives, improves immunity of organisms.Lactobacillus-fermented further improves the palatability after agriculture produced by fermenting waste residues and provides sour taste, Lactic acid bacteria as it is main can Direct-Fed Microbials can reduce rumen ecology disease incidence, improve immunity.

The present invention also provides the preparation methods of the grain ration of the pet, comprising the following steps:

(1) the agriculture waste residue of no mildew is taken to remove impurity, it is useless that the decomposition bacterium culture medium after sterilizing is uniformly then sprayed onto agricultural On slag, agriculture waste residue moisture content is made to reach 10% ~ 50%, obtains fermentation bottom material 1；

(2) bacterium solution seed liquor is decomposed according to 5% inoculation of fermentation 1 mass of bottom material, fermented under the conditions of being placed in 25 ~ 32 DEG C, fermentation 1 ~ 3 It when add promotor, bacterium to be decomposed stops fermentation after growing fructification, agriculture waste residue after ferment and decomposes mushroom reality Body, i.e. fermentation bottom material 2；

(3) after being crushed fermentation bottom material 2, lactic acid bacteria culturing medium is added according to the amount of 0.2L/kg fermentation bottom material 2, then connects The lactobacillus solution of kind fermentation 2 mass 5% of bottom material, stirs evenly, and continuous liquid ferments 10 ~ 15 days, obtains base-material；

(4) base-material is uniformly mixed to obtain component 1 with dregs of beans, wheat bran, Siraitia grosvenorii root tuber powder, salt, edible oil；

(5) corn flour, wheat flour, mealy potato, salt are mixed, obtains component 2；

(6) meat, vitamin, minerals are mixed, pickles 30 ~ 60 minutes at normal temperature, obtains component 3；

(7) component 1, component 2 and component 3 are mixed, bakes 8 ~ 10h at 60 ~ 70 DEG C then to get pet grain ration is arrived；

The decomposition bacterium culture medium contains following weight substance: CoCl in terms of 100mL₂•6H₂O 0.002g、CuSO₄•5H₂O 0.002g、ZnSO₄•7H₂O 0.001g、FeSO₄•7H₂O 0.005g、KH₂PO₄ 0.1g、K₂HPO₄•3H₂O 0.05g、MgSO₄ 0.02g、MnCl₂•4H₂O 0.009g、CaCl₂0.05g, corn flour 1g, peptone 0.3g, pH 6.0；

The lactic acid bacteria culturing medium contains following weight substance: sucrose 2g, peptone 1g, NaCl 0.5g in terms of 100mL.

As a further improvement of the present invention, in the preparation method of the pet grain ration, the step of (2) fermentation be solid-state Fermentation.

As a further improvement of the present invention, the decomposition bacterium solution seed liquor is black fungus bacterial seed liquor, Inonotus obliquus Two kinds of combinations in seed liquor and grifola frondosus seed liquor, preferably black fungus bacterial seed liquor, the combination of grifola frondosus seed liquor or black wood One of ear bacterium seed liquor, the combination of Inonotus obliquus seed liquor.

The Inonotus obliquus seed liquor the preparation method is as follows:

(1) aseptically, original Inonotus obliquus is inoculated on the plate of Mea culture medium with loop-carrier, then will be put down Plate is put into constant incubator, and 10d is cultivated at relative humidity 90%, 25 DEG C of temperature, and the mycelia of Inonotus obliquus is discontented with plate, i.e., The Inonotus obliquus of activation；

(2) fungus block for taking out 4 ~ 5 block sizes about 1 ~ 2cm from the Inonotus obliquus of activation with inoculation shovel, is inoculated into fluid nutrient medium 1 In, then inoculated fluid nutrient medium is placed in constant temperature and humidity incubator, cultivates 3 at relative humidity 80%, 25 DEG C of temperature It after ~ 4 days, is transferred into constant-temperature table, cultivates 4 ~ 5 days at 28 DEG C of temperature, revolving speed 150r/min to get Inonotus obliquus is arrived Seed liquor；

The Mea culture medium is prepared in terms of 100mL by following weight substance: peptone 0.3g, fructus hordei germinatus leaching powder 3g, fine jade Rouge 1.5g, pH are natural；

The fluid nutrient medium 1 is prepared: KH in terms of 100mL by following weight substance₂PO₄ 0.1g、MgSO₄ 0.15g、 CaCl₂0.01g, yeast extract 0.1g, peptone 0.3g, glucose 2g, pH are natural.

The black fungus bacterial seed liquor the preparation method is as follows:

(1) the mycelia block that semen viciae fabae size is cut out in black fungus bacterial parent species test tube, is inoculated in PDA slant medium 1, at 25 DEG C Lower culture 7 days, the black fungus mycelia activated；

(2) the black fungus mycelia of activation is cut into small pieces, is inoculated into PDB liquid seed culture medium, liquid amount 150mL/ 500mL triangular flask, one bottle of the black fungus mycelium inoculation of a slant activation cultivate 4 under the conditions of 25 DEG C, 200r/min shaking table It, obtains black fungus bacterial seed liquor；

The PDA slant medium 1 is prepared in terms of 1000mL by following weight substance: potato 200g, glucose 20g, agar 20g, pH are natural；The preparation method of the PDA slant medium 1, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor 1；

(2) mixed liquor 1 is taken into filtrate with eight layers of filtered through gauze, the glucose and agar of above-mentioned quality is added in filtrate, obtains Mixed liquor 2；

(3) mixed liquor 2 is heated and is stirred continuously with small fire, after agar is completely dissolved, benefit is filled with water to 1000mL to get arriving PDA slant medium 1；

The PDB liquid seed culture medium is prepared in terms of 1000mL by following weight substance: potato 200g, grape Sugared 20g, pH are natural；The preparation method of the PDB liquid seed culture medium, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor A；

(2) mixed liquor A is taken into filtrate with eight layers of filtered through gauze, the glucose of above-mentioned quality is added in filtrate, obtains mixed liquor B；

(3) mixed liquid B benefit is filled with water to 1000mL to get PDB liquid seed culture medium is arrived.

The grifola frondosus seed liquor the preparation method is as follows:

(1) 1 piece of blade is taken in " grifola frondosus " fructification obtained from field acquisition, with alcohol wipe 2 ~ 3 times, is then cut into the grain of rice Big small bacteria block is inoculated at 2,25 DEG C of PDA slant medium and cultivates 8 ~ 12 days, after rejuvenation culture 1 ~ 2 time, culture 5 ~ 7 days, takes shape The uniform bacterium colony of state to get arrive grifola frondosus parent species；

(2) fungus block for being obtained diameter 1cm from grifola frondosus parent species using metal punch, is inoculated in fluid nutrient medium 2,25 DEG C, 5 ~ 7 days are cultivated in 150r/min shaking table to get to grifola frondosus seed liquor；

The PDA slant medium 2 is prepared in terms of 1000mL by following weight substance: potato 200g, glucose 20g, agar 18g, pH are natural；The preparation method of the PDA slant medium 2, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor a；

(2) mixed liquor a is taken into filtrate with eight layers of filtered through gauze, the glucose and agar of above-mentioned quality is added in filtrate, obtains Mixed liquor b；

(3) mixed liquor b small fire is heated and is stirred continuously, after agar is completely dissolved, benefit is filled with water to 1000mL to get arriving PDA slant medium 2；

The fluid nutrient medium 2 is prepared in terms of 1000mL by following weight substance: glucose 30g, peptone 2g, MgSO₄·7H₂O 1g、KH₂PO₄2g, corn pulp 15g.

The lactobacillus solution the preparation method is as follows:

By lactobacillus inoculum in MRS fluid nutrient medium, 4 ~ 5 days are cultivated at 37 DEG C to get lactobacillus solution is arrived；

The MRS fluid nutrient medium is prepared in terms of 1000mL by following weight substance: peptone 10g, beef extract 10g, Yeast extract 5g, K₂HPO₄2g, dibasic ammonium citrate 2g, sodium acetate 5g, glucose 20g, MgSO₄ 0.5g、MnSO₄ 0.25g、 Tween 80 1mL, pH are 6 ~ 7.

Technical principle of the invention:

In lactic acid bacteria liquid fermentation process, the exocellular polysaccharide that lactic acid bacteria generates can stimulate black fungus, grifola frondosus and Inonotus obliquus It is metabolized, improves three kinds of fungies to the degradation rate of agriculture waste residue；On the other hand, the polysaccharide that black fungus liquid state fermentation generates can promote Into grifola frondosus, the yield of Inonotus obliquus extracellular polyphenol substance and enzyme, facilitate the cellulose and half fiber in fungi decomposition waste residue Dimension element, improves the nutritional ingredient in fermented material；Meanwhile Blackfungus polyhexose can improve grifola frondosus and be suitable for agriculture produced by fermenting waste residues Property.

Tween 80, oleic acid can largely promote the production of the mycelium and exo polysaccharides intracellular of black fungus and Inonotus obliquus Amount, tween can antagonism fatty acid synthesis, Tween 80 have longer C18 chain, can be resolved by microbial enzyme (such as lipase) The presence of oleic acid, Tween 80 also induces lipase active, and the oleic acid from Tween 80 decomposition can promote mycelial life Long, Tween 80 can also promote black fungus bacterial and Inonotus obliquus to absorb nutriment more effectively from fermentation liquid to promote biology Growth.And fatty acid provides raw material in film forming process for triglycerides synthesis, and influences rouge acyl chain to a certain extent Saturation degree, arrange evacuation degree by changing phospholipid bilayer to increase the mobility of cell membrane, as a result lead to cell The permeability of film is changed, while promoting black fungus bacterial and Inonotus obliquus that can make full use of the nutriment in fermentation liquid More polysaccharide are discharged to extracellularly, result in the increase of intraor extracellular polysaccharide.

Tween 80 and oleic acid are added in different fermentation times, will affect thallus number and thallus secretion exocellular polysaccharide, It is at maximum up to balanced in order to secrete thallus number maximum and exocellular polysaccharide, by verification experimental verification, Tween 80 adds the time bottom of as The 1st ~ 2 day of 1 fermentation of material, oleic acid add best when the 2nd ~ 3 day that the time is the fermentation of bottom material 1.

Beneficial effects of the present invention:

1, the present invention is prepared into pet by fungi and lactobacillus-fermented lignin degrading using agriculture waste residue bagasse, manioc waste Grain ration improves the utilization rate of agriculture waste residue, increases the processing approach of agriculture waste residue, new preparation is provided for feed for pet Approach substantially increases the utilization rate of raw material, has good economic benefit.

2, the present invention is improved using black fungus, Inonotus obliquus and the agriculture waste residue of grifola frondosus three classes fungi solid state fermentation degradation The degradation rate of lignin and hemicellulose, remains more celluloses, improves fermentation efficiency, shorten fermentation time； The crude protein and polysaccharide that black fungus, Inonotus obliquus and grifola frondosus three classes mushroom entity and fermentation generate provide more for pet grain ration More nutrition recycles lactic acid bacteria to carry out the fragrance that fermentation improves grain ration, substantially improves the pet of agriculture produced by fermenting waste residues preparation The palatability of grain ration；

3, the present invention promotes black fungus, Inonotus obliquus and grifola frondosus thalli growth and cell inside/outside using Tween 80 and oleic acid The content of polysaccharide makes thallus secrete more lignin decomposition enzymes and polysaccharide, improves the degradation rate of lignin and mention for feed For more nutriments, fermentation efficiency is further improved.Lactic acid bacteria, black fungus, grifola frondosus and Inonotus obliquus are utilized simultaneously In the mutual utilization of fermentation process metabolite, mutual promoting action, further improves the degradation efficiency of agriculture waste residue and dote on The nutriment of object grain ration, so that pet grain ration prepared by the present invention has very big nutritive value.

Specific embodiment

Below with reference to embodiment, the invention will be further described.

Embodiment 1

(1) it takes the bagasse of no mildew to remove impurity, the decomposition bacterium culture medium after sterilizing is uniformly then sprayed onto agriculture waste residue On, so that agriculture waste residue moisture content is reached 10%, obtains fermentation bottom material 1；

(2) bacterium solution seed liquor is decomposed according to 5% inoculation of fermentation 1 mass of bottom material, solid state fermentation is carried out under the conditions of being placed in 32 DEG C, is sent out 3 parts of Tween 80 that volume fraction is 0.1% are added at ferment 3 days, bacterium to be decomposed stops fermentation after growing fructification, after obtaining fermentation Agriculture waste residue and decomposer fructification, i.e., fermentation bottom material 2；

The decomposer is black fungus, grifola frondosus combination, and inoculum concentration is respectively the 2.5% of fermentation 1 mass of bottom material.

(3) after being crushed fermentation bottom material 2, lactic acid bacteria culturing medium is added according to the amount of 0.2L/kg fermentation bottom material 2, so The lactobacillus solution of 2 mass 5% of inoculation fermentation bottom material afterwards, stirs evenly, and continuous liquid ferments 15 days, obtains base-material；

The lactic acid bacteria be Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus helveticus, lactobacillus plantarum mixed in equal amounts and obtain, Every gram contains viable count > 4 × 10⁸It is a.

(4) base-material is uniformly mixed with 5 parts of dregs of beans, 5 parts of wheat bran, 10 parts of Siraitia grosvenorii root tuber powder, 2 parts of salt, 2 parts of peanut oil Obtain component 1；

(5) by 2 parts of corn flour, 5 parts of wheat flour, 2 parts of mealy potato, 4 parts of yellow meal worm powder, 5 parts of bone meal, 1 part of salt is mixed to get group Divide 2；

(6) by 20 parts of meat, 10 parts of vitamin, 10 parts of minerals mixing, room temperature is 20 minutes marinated, obtains component 3；

(7) it is that 3:4:3 is mixed according to component 1, component 2,3 mass ratio of component, bakes 10h at 60 DEG C to get pet is arrived Grain ration；

The meat is pork, beef, chicken mixed in equal amounts are prepared；

The vitamin be vitamin B, vitamin B12, vitamin C mixed in equal amounts and obtain；

The minerals obtain for K, Ca, Fe mixed in equal amounts.

The decomposition bacterium culture medium contains following weight substance: CoCl in terms of 100mL₂•6H₂O 0.002g、CuSO₄• 5H₂O 0.002g、ZnSO₄•7H₂O 0.001g、FeSO₄•7H₂O 0.005g、KH₂PO₄ 0.1g、K₂HPO₄•3H₂O 0.05g、 MgSO₄ 0.02g、MnCl₂•4H₂O 0.009g、CaCl₂0.05g, corn flour 1g, peptone 0.3g, pH 6.0；

The lactic acid bacteria culturing medium contains following weight substance: sucrose 2g, peptone 1g, NaCl 0.5g in terms of 100mL.

The black fungus bacterial seed liquor the preparation method is as follows:

(1) the mycelia block that semen viciae fabae size is cut out in black fungus bacterial parent species test tube, is inoculated in PDA slant medium 1, at 25 DEG C Lower culture 7 days, the black fungus mycelia activated；

(2) the black fungus mycelia of activation is cut into small pieces, is inoculated into PDB liquid seed culture medium, liquid amount 150mL/ 500mL triangular flask, one bottle of the black fungus mycelium inoculation of a slant activation cultivate 4 under the conditions of 25 DEG C, 200r/min shaking table It, obtains black fungus bacterial seed liquor；

The PDA slant medium 1 is prepared in terms of 1000mL by following weight substance: potato 200g, glucose 20g, agar 20g, pH are natural；The preparation method of the PDA slant medium 1, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor 1；

(2) mixed liquor 1 is taken into filtrate with eight layers of filtered through gauze, the glucose and agar of above-mentioned quality is added in filtrate, obtains Mixed liquor 2；

(3) mixed liquor 2 is heated and is stirred continuously with small fire, after agar is completely dissolved, benefit is filled with water to 1000mL to get arriving PDA slant medium 1；

The PDB liquid seed culture medium is prepared in terms of 1000mL by following weight substance: potato 200g, grape Sugared 20g, pH are natural；The preparation method of the PDB liquid seed culture medium, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor A；

(2) mixed liquor A is taken into filtrate with eight layers of filtered through gauze, the glucose of above-mentioned quality is added in filtrate, obtained you and mix Liquid B；

(3) mixed liquid B benefit is filled with water to 1000mL to get PDB liquid seed culture medium is arrived.

The grifola frondosus seed liquor the preparation method is as follows:

(1) 1 piece of blade is taken in " grifola frondosus " fructification obtained from field acquisition, with alcohol wipe 2 ~ 3 times, is then cut into the grain of rice Big small bacteria block is inoculated at 2,25 DEG C of PDA slant medium and cultivates 8 days, after rejuvenation culture 1 time, culture 5 days, takes form uniform Bacterium colony to get arrive grifola frondosus parent species；

(2) fungus block for being obtained diameter 1cm from grifola frondosus parent species using metal punch, is inoculated in fluid nutrient medium 2,25 DEG C, 5 days are cultivated in 150r/min shaking table to get to grifola frondosus seed liquor；

The PDA slant medium 2 is prepared in terms of 1000mL by following weight substance: potato 200g, glucose 20g, agar 18g, pH are natural；The preparation method of the PDA slant medium 2, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor a；

(2) mixed liquor a is taken into filtrate with eight layers of filtered through gauze, the glucose and agar of above-mentioned quality is added in filtrate, obtains Mixed liquor b；

(3) mixed liquor b small fire is heated and is stirred continuously, after agar is completely dissolved, benefit is filled with water to 1000mL to get arriving PDA slant medium 2；

The fluid nutrient medium 2 is prepared in terms of 1000mL by following weight substance: glucose 30g, peptone 2g, MgSO₄·7H₂O 1g、KH₂PO₄2g, corn pulp 15g.

The lactobacillus solution the preparation method is as follows:

By lactobacillus inoculum in MRS fluid nutrient medium, 4 days are cultivated at 37 DEG C to get lactobacillus solution is arrived；

The MRS fluid nutrient medium is prepared in terms of 1000mL by following weight substance: peptone 10g, beef extract 10g, Yeast extract 5g, K₂HPO₄2g, dibasic ammonium citrate 2g, sodium acetate 5g, glucose 20g, MgSO₄ 0.5g、MnSO₄ 0.25g、 Tween 80 1mL, pH 6.

Embodiment 2

(1) it takes the manioc waste of no mildew to remove impurity, the decomposition bacterium culture medium after sterilizing is uniformly then sprayed onto agriculture waste residue On, so that agriculture waste residue moisture content is reached 50%, obtains fermentation bottom material 1；

(2) bacterium solution seed liquor is decomposed according to 5% inoculation of fermentation 1 mass of bottom material, solid state fermentation is carried out under the conditions of being placed in 25 DEG C, is sent out 1 part of oleic acid that volume fraction is 0.5% is added at ferment 1 day, bacterium to be decomposed stops fermentation after growing fructification, after being fermented Agriculture waste residue and decomposer fructification, i.e. fermentation bottom material 2；

The decomposer is black fungus, Inonotus obliquus combination, and inoculum concentration is respectively the 2.5% of fermentation 1 mass of bottom material.

(3) after being crushed fermentation bottom material 2, lactic acid bacteria culturing medium is added according to the amount of 0.2L/kg fermentation bottom material 2, so The lactobacillus solution of 2 mass 5% of inoculation fermentation bottom material afterwards, stirs evenly, and continuous liquid ferments 10 days, obtains base-material；

The lactic acid bacteria be Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus helveticus, lactobacillus plantarum mixed in equal amounts and obtain, Every gram contains viable count > 4 × 10⁸It is a.

(4) by base-material and 10 parts of dregs of beans, 10 parts of wheat bran, 15 parts of Siraitia grosvenorii root tuber powder, 5 parts of salt, peanut oil or olive oil 5 Part is uniformly mixed and obtains component 1；

(5) by 5 parts of corn flour, 8 parts of wheat flour, 5 parts of mealy potato, 7 parts of yellow meal worm powder, 10 parts of bone meal, 3 parts of salt are mixed to get group Divide 2；

(6) by 50 parts of meat, 5 parts of vitamin, 5 parts of minerals mixing, room temperature is 40 minutes marinated, obtains component 3；

(7) it is that 4:4:2 is mixed according to component 1, component 2,3 mass ratio of component, bakes 8h at 70 DEG C to get pet mouthful is arrived Grain；

The meat be chicken, duck, flesh of fish mixed in equal amounts and obtain；

The vitamin be vitamin B, vitamin E, vitamin B12, vitamin C mixed in equal amounts and obtain；

The minerals obtain for K, Na, Ca, Zn, Fe mixed in equal amounts.

The decomposition bacterium culture medium contains following weight substance: CoCl in terms of 100mL₂•6H₂O 0.002g、CuSO₄• 5H₂O 0.002g、ZnSO₄•7H₂O 0.001g、FeSO₄•7H₂O 0.005g、KH₂PO₄ 0.1g、K₂HPO₄•3H₂O 0.05g、 MgSO₄ 0.02g、MnCl₂•4H₂O 0.009g、CaCl₂0.05g, corn flour 1g, peptone 0.3g, pH 6.0；

The lactic acid bacteria culturing medium contains following weight substance: sucrose 2g, peptone 1g, NaCl 0.5g in terms of 100mL.

The Inonotus obliquus seed liquor the preparation method is as follows:

(1) aseptically, original Inonotus obliquus is inoculated on the plate of Mea culture medium with loop-carrier, then will be put down Plate is put into constant incubator, and 10d is cultivated at relative humidity 90%, 25 DEG C of temperature, and the mycelia of Inonotus obliquus is discontented with plate, i.e., The Inonotus obliquus of activation；

(2) fungus block for taking out 5 block sizes about 1 ~ 2cm from the Inonotus obliquus of activation with inoculation shovel, is inoculated into fluid nutrient medium 1 In, then inoculated fluid nutrient medium is placed in constant temperature and humidity incubator, cultivates 4 at relative humidity 80%, 25 DEG C of temperature It after it, is transferred into constant-temperature table, cultivates 5 days at 28 DEG C of temperature, revolving speed 150r/min to get Inonotus obliquus seed is arrived Liquid；

The Mea culture medium is prepared in terms of 100mL by following weight substance: peptone 0.3g, fructus hordei germinatus leaching powder 3g, fine jade Rouge 1.5g, pH are natural；

The fluid nutrient medium 1 is prepared: KH in terms of 100mL by following weight substance₂PO₄ 0.1g、MgSO₄ 0.15g、 CaCl₂0.01g, yeast extract 0.1g, peptone 0.3g, glucose 2g, pH are natural.

The black fungus bacterial seed liquor the preparation method is as follows:

(1) the mycelia block that semen viciae fabae size is cut out in black fungus bacterial parent species test tube, is inoculated in PDA slant medium 1, at 25 DEG C Lower culture 7 days, the black fungus mycelia activated；

(2) the black fungus mycelia of activation is cut into small pieces, is inoculated into PDB liquid seed culture medium, liquid amount 150mL/ 500mL triangular flask, one bottle of the black fungus mycelium inoculation of a slant activation cultivate 4 under the conditions of 25 DEG C, 200r/min shaking table It, obtains black fungus bacterial seed liquor；

The PDA slant medium 1 is prepared in terms of 1000mL by following weight substance: potato 200g, glucose 20g, agar 20g, pH are natural；The preparation method of the PDA slant medium 1, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor 1；

(2) mixed liquor 1 is taken into filtrate with eight layers of filtered through gauze, the glucose and agar of above-mentioned quality is added in filtrate, obtains Mixed liquor 2；

(3) mixed liquor 2 is heated and is stirred continuously with small fire, after agar is completely dissolved, benefit is filled with water to 1000mL to get arriving PDA slant medium 1；

The PDB liquid seed culture medium is prepared in terms of 1000mL by following weight substance: potato 200g, grape Sugared 20g, pH are natural；The preparation method of the PDB liquid seed culture medium, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor A；

(2) mixed liquor A is taken into filtrate with eight layers of filtered through gauze, the glucose of above-mentioned quality is added in filtrate, obtains mixed liquor B；

(3) mixed liquid B benefit is filled with water to 1000mL to get PDB liquid seed culture medium is arrived.

The lactobacillus solution the preparation method is as follows:

By lactobacillus inoculum in MRS fluid nutrient medium, 5 days are cultivated at 37 DEG C to get lactobacillus solution is arrived；

The MRS fluid nutrient medium is prepared in terms of 1000mL by following weight substance: peptone 10g, beef extract 10g, Yeast extract 5g, K₂HPO₄2g, dibasic ammonium citrate 2g, sodium acetate 5g, glucose 20g, MgSO₄ 0.5g、MnSO₄ 0.25g、 Tween 80 1mL, pH 7.

Embodiment 3

(1) it takes the bagasse of no mildew to remove impurity, the decomposition bacterium culture medium after sterilizing is uniformly then sprayed onto agriculture waste residue On, so that agriculture waste residue moisture content is reached 30%, obtains fermentation bottom material 1；

(2) bacterium solution seed liquor is decomposed according to 5% inoculation of fermentation 1 mass of bottom material, solid state fermentation is carried out under the conditions of being placed in 30 DEG C, is sent out 2 parts of Tween 80 that volume fraction is 0.4% are added at ferment 2 days, bacterium to be decomposed stops fermentation after growing fructification, after obtaining fermentation Agriculture waste residue and decomposer fructification, i.e., fermentation bottom material 2；

The decomposer is black fungus, Inonotus obliquus combination, and inoculum concentration is respectively the 2.5% of fermentation 1 mass of bottom material.

(3) after 2 row of bottom material that will ferment crushes, lactic acid bacteria culturing medium is added according to the amount of 0.2L/kg fermentation bottom material 2, then The lactobacillus solution of 2 mass 5% of inoculation fermentation bottom material, stirs evenly, and continuous liquid ferments 12 days, obtains base-material；

The lactic acid bacteria be Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus helveticus, lactobacillus plantarum mixed in equal amounts and obtain, Every gram contains viable count > 4 × 10⁸It is a.

(4) base-material is uniformly mixed with 6 parts of dregs of beans, 8 parts of wheat bran, 12 parts of Siraitia grosvenorii root tuber powder, 4 parts of salt, 3 parts of peanut oil Obtain component 1；

(5) by 3 parts of corn flour, 6 parts of wheat flour, 4 parts of mealy potato, 5 parts of yellow meal worm powder, 7 parts of bone meal, 2 parts of salt are mixed to get group Divide 2；

(6) by 30 parts of meat, 8 parts of vitamin, 6 parts of minerals mixing, room temperature is 30 minutes marinated, obtains component 3；

(7) it is that 4:5:1 is mixed according to component 1, component 2,3 mass ratio of component, bakes 10h at 65 DEG C to get pet is arrived Grain ration；

The meat be pork, mutton, the flesh of fish, duck mixed in equal amounts and obtain；

The vitamin be vitamin B, vitamin E, vitamin C mixed in equal amounts and obtain；

The minerals obtain for K, Na, Ca, Zn mixed in equal amounts.

The decomposition bacterium culture medium contains following weight substance: CoCl in terms of 100mL₂•6H₂O 0.002g、CuSO₄• 5H₂O 0.002g、ZnSO₄•7H₂O 0.001g、FeSO₄•7H₂O 0.005g、KH₂PO₄ 0.1g、K₂HPO₄•3H₂O 0.05g、 MgSO₄ 0.02g、MnCl₂•4H₂O 0.009g、CaCl₂0.05g, corn flour 1g, peptone 0.3g, pH 6.0；

The lactic acid bacteria culturing medium contains following weight substance: sucrose 2g, peptone 1g, NaCl 0.5g in terms of 100mL.

The Inonotus obliquus seed liquor the preparation method is as follows:

(1) aseptically, original Inonotus obliquus is inoculated on the plate of Mea culture medium with loop-carrier, then will be put down Plate is put into constant incubator, and 10d is cultivated at relative humidity 90%, 25 DEG C of temperature, and the mycelia of Inonotus obliquus is discontented with plate, i.e., The Inonotus obliquus of activation；

(2) fungus block for taking out 4 ~ 5 block sizes about 1 ~ 2cm from the Inonotus obliquus of activation with inoculation shovel, is inoculated into fluid nutrient medium 1 In, then inoculated fluid nutrient medium is placed in constant temperature and humidity incubator, cultivates 3 at relative humidity 80%, 25 DEG C of temperature It after ~ 4 days, is transferred into constant-temperature table, cultivates 4 ~ 5 days at 28 DEG C of temperature, revolving speed 150r/min to get Inonotus obliquus is arrived Seed liquor；

The Mea culture medium is prepared in terms of 100mL by following weight substance: peptone 0.3g, fructus hordei germinatus leaching powder 3g, fine jade Rouge 1.5g, pH are natural；

The fluid nutrient medium 1 is prepared: KH in terms of 100mL by following weight substance₂PO₄ 0.1g、MgSO₄ 0.15g、 CaCl₂0.01g, yeast extract 0.1g, peptone 0.3g, glucose 2g, pH are natural.

The black fungus bacterial seed liquor the preparation method is as follows:

(1) the mycelia block that semen viciae fabae size is cut out in black fungus bacterial parent species test tube, is inoculated in PDA slant medium 1, at 25 DEG C Lower culture 7 days, the black fungus mycelia activated；

(2) the black fungus mycelia of activation is cut into small pieces, is inoculated into PDB liquid seed culture medium, liquid amount 150mL/ 500mL triangular flask, one bottle of the black fungus mycelium inoculation of a slant activation cultivate 4 under the conditions of 25 DEG C, 200r/min shaking table It, obtains black fungus bacterial seed liquor；

The PDA slant medium 1 is prepared in terms of 1000mL by following weight substance: potato 200g, glucose 20g, agar 20g, pH are natural；The preparation method of the PDA slant medium 1, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor 1；

(2) mixed liquor 1 is taken into filtrate with eight layers of filtered through gauze, the glucose and agar of above-mentioned quality is added in filtrate, obtains Mixed liquor 2；

(3) mixed liquor 3 is heated and is stirred continuously with small fire, after agar is completely dissolved, benefit is filled with water to 1000mL to get arriving PDA slant medium 1；

The PDB liquid seed culture medium is prepared in terms of 1000mL by following weight substance: potato 200g, grape Sugared 20g, pH are natural；The preparation method of the PDB liquid seeds culture, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor A；

(2) mixed liquor A is taken into filtrate with eight layers of filtered through gauze, the glucose of above-mentioned quality is added in filtrate, obtains mixed liquor B；

(3) mixed liquid B benefit is filled with water to 1000mL to get PDB liquid seed culture medium is arrived.

The lactobacillus solution the preparation method is as follows:

By lactobacillus inoculum in MRS fluid nutrient medium, 4 ~ 5 days are cultivated at 37 DEG C to get lactobacillus solution is arrived；

The MRS fluid nutrient medium is prepared in terms of 1000mL by following weight substance: peptone 10g, beef extract 10g, Yeast extract 5g, K₂HPO₄2g, dibasic ammonium citrate 2g, sodium acetate 5g, glucose 20g, MgSO₄ 0.5g、MnSO₄ 0.25g、 Tween 80 1mL, pH are 6 ~ 7.

Test example 1

Pet grain ration prepared by the embodiment of the present invention 1 is fed into 30 pet cats with common commercially available pet cat food respectively, it is continuous to feed It supports 3 months, sick to pet cat total number of elements, monthly weight gain value (average value)

It is counted, the results are shown in Table 1.

1 pet cat growing state table of table

Test example 2

Pet grain ration prepared by the embodiment of the present invention 2 is fed into 50 hamsters with common commercially available hamster grain ration respectively, it is continuous to feed It supports 3 months, the total amount of feed weekly (average value) of hamster, weight gain value (average value), sick total degree is counted, unite Meter the results are shown in Table 2.

2 hamster growing state table of table

Test example 3

Pet grain ration prepared by the embodiment of the present invention 3 is fed into 20 pet dogs with common commercially available dog food respectively, continuously feeds 6 A month, sick total degree is caused to the infection of pet dog germ and monthly body weight increase numerical value (average value) counts, as a result It is shown in Table 3 and table 4.

The sick number of 3 pet dog of table

4 pet dog of table monthly weight gain value

By 1 ~ 3 all data of test example the results show that compared to commercially available normal diet, pet grain ration prepared by the present invention can have Effect improves the immunity of organisms of pet, promotes pet growth.

Above embodiments are only exemplary embodiment of the present invention, are not used in the limitation present invention, protection scope of the present invention It is defined by the claims.Those skilled in the art can make various repair to the present invention within the spirit and scope of the present invention Change or equivalent replacement, this modification or equivalent replacement also should be regarded as being within the scope of the present invention.

Claims (10)

1. a kind of pet grain ration, which is characterized in that including component 1, component 2 and component 3；The component 1 is by following parts by weight Number is than substance by being prepared after decomposer and lactobacillus-fermented: 20 ~ 40 parts of agriculture waste residue, 1 ~ 3 part of promotor, dregs of beans 5 ~ 10 Part, 5 ~ 10 parts of wheat bran, 10 ~ 15 parts of Siraitia grosvenorii root tuber powder, 2 ~ 5 parts of salt, 2 ~ 5 parts of edible oil；The component 2 includes following heavy Measure portion rate substance: 2 ~ 5 parts of corn flour, 5 ~ 8 parts of wheat flour, 2 ~ 5 parts of mealy potato, 1 ~ 3 part of salt；The component 3 is by following Weight fraction ratio material composition: 20 ~ 50 parts of meat, 5 ~ 10 parts of vitamin, 5 ~ 10 parts of minerals；

The agriculture waste residue is one of bagasse, manioc waste；

The promotor is one of Tween 80, oleic acid, and volume fraction is 0.1 ~ 0.5%；

The edible oil is one of peanut oil, olive oil；

The meat is one of pork, beef, mutton, the flesh of fish, chicken, duck or a variety of；

The vitamin is vitamin B, vitamin E, vitamin B12, one or more in vitamin C；

The minerals are one or more in K, Na, Ca, Zn, Fe.

2. pet grain ration according to claim 1, which is characterized in that the component 1, component 2,3 mass ratio of component are (3~4):(4~5):(1~3)。

3. the pet grain ration according to claim 1, which is characterized in that the lactic acid bacteria is rhamnose cream bar Bacterium, Lactobacillus casei, Lactobacillus helveticus, lactobacillus plantarum mixed in equal amounts and obtain, every gram contain viable count > 4 × 10⁸It is a.

4. such as the preparation method of any pet grain ration of claim 1 ~ 3, which comprises the following steps:

(1) the agriculture waste residue of no mildew is taken to remove impurity, it is useless that the decomposition bacterium culture medium after sterilizing is uniformly then sprayed onto agricultural On slag, agriculture waste residue moisture content is made to reach 10% ~ 50%, obtains fermentation bottom material 1；

(2) bacterium solution seed liquor is decomposed according to 5% inoculation of fermentation 1 mass of bottom material, fermented under the conditions of being placed in 25 ~ 32 DEG C, fermentation 1 ~ 3 It when add promotor, bacterium to be decomposed stops fermentation after growing fructification, agriculture waste residue after ferment and decomposes mushroom reality Body, i.e. fermentation bottom material 2；

(3) after being crushed fermentation bottom material 2, lactic acid bacteria culturing medium is added according to the amount of 0.2L/kg fermentation bottom material 2, then connects The lactobacillus solution of kind fermentation 2 mass 5% of bottom material, stirs evenly, and continuous liquid ferments 10 ~ 15 days, obtains base-material；

(4) base-material is uniformly mixed to obtain component 1 with dregs of beans, wheat bran, Siraitia grosvenorii root tuber powder, salt, edible oil；

(5) corn flour, wheat flour, mealy potato, salt are mixed, obtains component 2；

(6) meat, vitamin, minerals are mixed, pickles 30 ~ 60 minutes at normal temperature, obtains component 3；

(7) component 1, component 2 and component 3 are mixed, bakes 8 ~ 10h at 60 ~ 70 DEG C then to get pet grain ration is arrived；

The decomposition bacterium culture medium contains following weight substance: CoCl in terms of 100mL₂•6H₂O 0.002g、CuSO₄•5H₂O 0.002g、ZnSO₄•7H₂O 0.001g、FeSO₄•7H₂O 0.005g、KH₂PO₄ 0.1g、K₂HPO₄•3H₂O 0.05g、MgSO₄ 0.02g、MnCl₂•4H₂O 0.009g、CaCl₂0.05g, corn flour 1g, peptone 0.3g, pH 6.0；

The lactic acid bacteria culturing medium contains following weight substance: sucrose 2g, peptone 1g, NaCl 0.5g in terms of 100mL.

5. the preparation method of pet grain ration according to claim 4, which is characterized in that the fermentation of the step (2) is Solid state fermentation.

6. the preparation method of pet grain ration according to claim 4, which is characterized in that the lactobacillus solution preparation Method is as follows:

By lactobacillus inoculum in MRS fluid nutrient medium, 4 ~ 5 days are cultivated at 37 DEG C to get lactobacillus solution is arrived；

The MRS fluid nutrient medium is prepared in terms of 1000mL by following weight substance: peptone 10g, beef extract 10g, Yeast extract 5g, K₂HPO₄2g, dibasic ammonium citrate 2g, sodium acetate 5g, glucose 20g, MgSO₄ 0.5g、MnSO₄ 0.25g、 Tween 80 1mL, pH are 6 ~ 7.

7. the preparation method of pet grain ration according to claim 4, which is characterized in that the decomposition bacterium solution seed liquor is Two kinds of combinations in black fungus bacterial seed liquor, Inonotus obliquus seed liquor and grifola frondosus seed liquor, preferably black fungus bacterial seed liquor, The combination of grifola frondosus seed liquor or one of black fungus bacterial seed liquor, the combination of Inonotus obliquus seed liquor.

8. the preparation method of pet grain ration according to claim 7, which is characterized in that the Inonotus obliquus seed liquor system Preparation Method is as follows:

(1) aseptically, original Inonotus obliquus is inoculated on the plate of Mea culture medium with loop-carrier, then will be put down Plate is put into constant incubator, and 10d is cultivated at relative humidity 90%, 25 DEG C of temperature, and the mycelia of Inonotus obliquus is discontented with plate, i.e., The Inonotus obliquus of activation；

(2) fungus block for taking out 4 ~ 5 block sizes about 1 ~ 2cm from the Inonotus obliquus of activation with inoculation shovel, is inoculated into fluid nutrient medium 1 In, then inoculated fluid nutrient medium is placed in constant temperature and humidity incubator, cultivates 3 at relative humidity 80%, 25 DEG C of temperature It after ~ 4 days, is transferred into constant-temperature table, cultivates 4 ~ 5 days at 28 DEG C of temperature, revolving speed 150r/min to get Inonotus obliquus is arrived Seed liquor；

The Mea culture medium is prepared in terms of 100mL by following weight substance: peptone 0.3g, fructus hordei germinatus leaching powder 3g, fine jade Rouge 1.5g, pH are natural；

The fluid nutrient medium 1 is prepared: KH in terms of 100mL by following weight substance₂PO₄ 0.1g、MgSO₄ 0.15g、 CaCl₂0.01g, yeast extract 0.1g, peptone 0.3g, glucose 2g, pH are natural.

9. the preparation method of pet grain ration according to claim 7, which is characterized in that the black fungus bacterial seed liquor system Preparation Method is as follows:

(1) the mycelia block that semen viciae fabae size is cut out in black fungus bacterial parent species test tube, is inoculated in PDA slant medium 1, at 25 DEG C Lower culture 7 days, the black fungus mycelia activated；

(2) the black fungus mycelia of activation is cut into small pieces, is inoculated into PDB liquid seed culture medium, liquid amount 150mL/ 500mL triangular flask, one bottle of the black fungus mycelium inoculation of a slant activation cultivate 4 under the conditions of 25 DEG C, 200r/min shaking table It, obtains black fungus bacterial seed liquor；

The PDA slant medium 1 is prepared in terms of 1000mL by following weight substance: potato 200g, glucose 20g, agar 20g, pH are natural；The preparation method of the PDA slant medium 1, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor 1；

(2) mixed liquor 1 is taken into filtrate with eight layers of filtered through gauze, the glucose and agar of above-mentioned quality is added in filtrate, obtains Mixed liquor 2；

(3) mixed liquor 2 is heated and is stirred continuously with small fire, after agar is completely dissolved, benefit is filled with water to 1000mL to get arriving PDA slant medium 1；

The PDB liquid seed culture medium is prepared in terms of 1000mL by following weight substance: potato 200g, grape Sugared 20g, pH are natural；The preparation method of the PDB liquid seed culture medium, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor A；

(2) mixed liquor A is taken into filtrate with eight layers of filtered through gauze, the glucose of above-mentioned quality is added in filtrate, obtains mixed liquor B；

(3) mixed liquid B benefit is filled with water to 1000mL to get PDB liquid seed culture medium is arrived.

10. the preparation method of pet grain ration according to claim 7, which is characterized in that the grifola frondosus seed liquor system Preparation Method is as follows:

(1) 1 piece of blade is taken in " grifola frondosus " fructification obtained from field acquisition, with alcohol wipe 2 ~ 3 times, is then cut into the grain of rice Big small bacteria block is inoculated on PDA slant medium 2, is cultivated 8 ~ 12 days at 25 DEG C, after rejuvenation culture 1 ~ 2 time, culture 5 ~ 7 days, is taken The uniform bacterium colony of form to get arrive grifola frondosus parent species；

(2) fungus block for being obtained diameter 1cm from grifola frondosus parent species using metal punch, is inoculated in fluid nutrient medium 2,25 DEG C, 5 ~ 7 days are cultivated in 150r/min shaking table to get to grifola frondosus seed liquor；

The PDA slant medium 2 is prepared in terms of 1000mL by following weight substance: potato 200g, glucose 20g, agar 18g, pH are natural；The preparation method of the PDA slant medium 2, comprising the following steps:

(1) it weighs after 200g potato first cleans peeling and is cut into small pieces, add 1000mL boiling rotten, obtain mixed liquor a；

(2) mixed liquor a is taken into filtrate with eight layers of filtered through gauze, the glucose and agar of above-mentioned quality is added in filtrate, obtains Mixed liquor b；

(3) mixed liquor b small fire is heated and is stirred continuously, after agar is completely dissolved, benefit is filled with water to 1000mL to get arriving PDA slant medium 2；

The fluid nutrient medium 2 is prepared in terms of 1000mL by following weight substance: glucose 30g, peptone 2g, MgSO₄·7H₂O 1g、KH₂PO₄2g, corn pulp 15g.