CN101356952B - Use of golden mushroom in animal production - Google Patents
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- CN101356952B CN101356952B CN2007101198950A CN200710119895A CN101356952B CN 101356952 B CN101356952 B CN 101356952B CN 2007101198950 A CN2007101198950 A CN 2007101198950A CN 200710119895 A CN200710119895 A CN 200710119895A CN 101356952 B CN101356952 B CN 101356952B
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Abstract
The invention relates to an application of flammulina velutipe powders in animal production, which belongs to the field of microbe making. The flammulina velutiper powders adopt the following fermentation medium formula with Flammulina velutipes (Fr.)Sing.As5.632 as the production strains to carry out the liquid submerged fermentation: the formula of the medium per liter is as follows: 20g of bran, 5g of corns, 10g of sucrose, 50g of soybean powders, 1g of monopotassium phosphate and 0.5g of magnesium sulfate are added with water to the constant volume of 1 liter; the inoculum concentration is 5 to 10 percent, the medium volume accounts for 60 to 70 percent of the volume of a fermentation tank, the fermentation temperature is 22 to 25 DEG C, the ventilation volume is 1:0.04 to 1.0, the medium is scaled up by passing through a shake flask, a liquid seeding tank and a liquid fermentation tank, and the fermentation time of each stage is 72 to 120 hours; the fermented fermentation liquid is filtered and concentrated, then the concentrated liquid and the filtering wet mycelium are dried and crashed. The flammulina velutiper powders which are produced by the invention can be widely applied to the feed industry, and can markedly improve the immunologic functions of the domestic animals and reduce the amount of antibiotic without any side effect, while improving the growth of the animals, and reduces the death rate.
Description
Technical field
The present invention relates to the application of golden mushroom in animal produces, refer to the fermentative production and the application in animal produces thereof of the golden mushroom that a kind of industrialization liquid fermentation process is produced especially, microorganism belonging to genus is made the field.
Background technology
Along with global economic integration, food safety has become the important public health problem of a global challenge and the whole world.The safety of animal-derived food (meat, egg, milk, aquatic products and goods thereof) is the focus that the whole world is paid close attention to, and wherein microbiotic and chemicals residue problem are one of important factors that influences animal-derived food safety.The life-time service microbiotic can produce problems such as resistance and environmental pollution, animal cultivation is too dependent on antibiotic etc, many problems have been produced, as causing drug residue behind the animal feeding antibiotic, cause the Resistant strain diffusion, animal, people and ecotope are caused serious harm, cause the animal flora imbalance, the immunizing power that suppresses animal, the secondary superinfection.The high amount of drug of Shi Yonging is absorbed by the body by food chain simultaneously, has produced bad consequence.Residue problem not only influences Economic development and social stability, also has a strong impact on product trade war internationally.Developed country attaches great importance to feed hygiene safety, the residual harm of medicine.Just begin the restricted part microbiotic as fodder additives as far back as Switzerland in 1986, European Union adds microbiotic with completely forbidding in 2006 in feed.State such as the U.S., Japan also constantly strengthens the restriction to feeding antibiotic.
Another subject matter that present China breeding production faces is that Animal diseases are not effectively prevented and controlled, so that can not bring into play the production potential of animal varieties and feed to greatest extent.According to statistics, the mortality ratio of China livestock and poultry surpasses the world average level far away 8～20%.Wherein bacterium and virus disease are the formidable enemies of harm aquaculture.The immunosuppressive virus infection of popular pig, chicken makes that this problem is more complicated in recent years.With the Swine Production is example, and multisystemic wasting syndrome makes insecondary infection rise one after another after the wean that immunosuppressive virus infections such as pig breeding and respiratory complication (PRRS) virus, pig II type PCV-II cause.Horse Garrick (MD), infectious bursa of Fabricius (IBD), generally infecting of chicken infectious anemia immunosuppressive virus such as (CIA) have become the important diseases that influence is raised chickens and produced.For the immunosuppressive virus disease, how to select suitable pharmacological agent is the problem that dealer's headache is cultured in order always, and the pharmacological agent of virus disease also is difficult to prove effective, and must carry out immunization and be prevented.But in process of production, the immuning failure phenomenon of animal takes place again often, how to guarantee that immunity successfully is the problem that must consider in producing.All the time, people generally add microbiotic in feed takes place with control disease, promote growth of animal, but antibiotic abuse has caused resistance and residue problem.Therefore, the development of microbiotic substitute will become one of main flow direction of feedstuff industry development from now on.And enhance immunity power is the key of enantiopathy viral disease and basic.Polysaccharide is now just paid close attention to by people as a kind of biological non-specific immunity promotor.
Summary of the invention
The primary technical problem that the present invention will solve provides a kind of golden mushroom of producing with the industrialization liquid fermentation process.
Second technical problem that the present invention will solve provides the manufacturing golden mushroom liquid fermentation producing technology that is suitable for large-scale industrial production.
The 3rd technical problem that the present invention will solve provides the application of golden mushroom aspect the raising breeding performonce fo animals.
For achieving the above object, the present invention is by the following technical solutions:
A kind of golden mushroom, adopt the preparation of following method: with needle mushroom Flammulina velutipes (Fr.) Sing.As5.632 (purchasing) in Institute of Microorganism, Academia Sinica as the production bacterial classification, adopt following fermentative medium formula, carry out liquid submerged fermentation:
Every liter of culture medium prescription is as follows: wheat bran 20g, corn 5g, sucrose 10g, bean powder 50g, potassium primary phosphate 1g, sal epsom 0.5g), add water and be settled to 1 liter, (without adjust pH);
Inoculum size is 5-10%, and culture volume accounts for the 60-70% of fermentor tank volume, and leavening temperature is 22-25 ℃, and air flow is 1:0.04-1.0, amplifies step by step through shaking bottle, liquid seeds jar, liquid fermentation tank, and every grade of fermentation time is 72-120 hours;
Fermentation secondary fermentation liquid after filtration, filtered liquid is concentrated, concentrated solution with filter wet mycelium and carry out drying, pulverize.
A kind of product that contains golden mushroom adopts method for preparing, after filtered liquid concentrates, adds carrier in concentrated solution and filtration wet mycelium, carries out drying, mixes powder after the pulverizing.
Described carrier is the corn cob fine powder, and the 80-100 order is pressed 1:1 (per 1 liter of concentrated solution adds 1 kilogram of carrier) with concentrated solution and added.
A kind of manufacturing golden mushroom liquid fermentation producing technology, with needle mushroom Flammulina velutipes (Fr.) Sing.As5.632 (purchasing) in Institute of Microorganism, Academia Sinica as producing bacterial classification, adopt following fermentative medium formula, carry out liquid submerged fermentation:
Every liter of culture medium prescription is as follows: wheat bran 20g, corn 5g, sucrose 10g, bean powder 50g, potassium primary phosphate 1g, sal epsom 0.5g without adjust pH, add water and are settled to 1 liter;
Inoculum size is 5-10%, and culture volume accounts for the 60-70% of fermentor tank volume, and leavening temperature is 22-25 ℃, and air flow is 1:0.04-1.0, amplifies step by step through shaking bottle, liquid seeds jar, liquid fermentation tank, and every grade of fermentation time is 72-120 hours; Fermentation secondary fermentation liquid after filtration, filtered liquid is concentrated adds carrier to concentrated solution with filtering in the wet mycelium, carries out drying, pulverizes.
Being filtered into of described fermentation secondary fermentation liquid through Plate Filtration.
Described filtered liquid is concentrated, for the thin film concentration method with 1/8 to 1/10 of filtered liquid simmer down to original volume.
The application of golden mushroom aspect the raising breeding performonce fo animals.Described animal is livestock and poultry and aquatic animal.The present invention proves first that by a large amount of experimentation on animalies golden mushroom can effectively promote animal to produce, thereby can be as novel fodder additive.
The invention has the beneficial effects as follows: (1) method of the present invention can significantly improve the content of polysaccharide in the golden mushroom; (2) carrier that is added in the production technique is a corn cob, is of value to the application in animal produces; (3) golden mushroom is in improving breeding performonce fo animals and have outstanding effect aspect Immune System for Animal; (4) golden mushroom of Sheng Chaning, but widespread use feedstuff industry, golden mushroom can significantly improve immunologic function of livestock and birds as the novel green fodder additives, improve disease resistance, can reduce antibiotic dosage, without any side effects, simultaneously can promote growth of animal, reduce mortality ratio.
Being described further below in conjunction with the drawings and specific embodiments, so that the public fully understands novelty of the present invention and creative place, is not limitation of the invention.All any technical characterictics of implementing according to disclosure of the present invention be equal to replacement, all belong within protection scope of the present invention.
Description of drawings
Fig. 1 is technological process of production figure of the present invention.
Embodiment
Embodiment 1: preparation contains the product of golden mushroom
Needle mushroom Flammulina velutipes (Fr.) Sing.As5.632 (purchasing in Institute of Microorganism, Academia Sinica) adopts the substratum of following proportioning as producing bacterial classification, carries out liquid submerged fermentation.
Every liter of culture medium prescription is as follows: wheat bran 20g, corn 5g, sucrose 10g, bean powder 50g, potassium primary phosphate 1g, sal epsom 0.5g without adjust pH, add water and are settled to 1 liter;
Inoculum size is 5-10%, and culture volume accounts for the 60-70% of fermentor tank volume, and leavening temperature is 22-25 ℃, and air flow is 1:0.04-1.0, and fermentation time is 96-120 hours, is specially:
(1) from inclined-plane picking bacterial classification inoculation in shaking bottle, 23-25 ℃, 96-120 hour;
(2) shake bottle and be amplified to 100L liquid seeds jar, 23-25 ℃, 72-96 hour;
(3) 100L liquid seeds jar is amplified to the 1000L liquid fermentation tank, and 23-25 ℃, 72-96 hour;
(4) the 1000L liquid fermentation tank is amplified to the 5000L liquid fermentation tank, and 23-25 ℃, 72-96 hour;
Final fermentor tank volume is 5000L, maximum loading amount is 70% of a fermentor tank volume, tunning through Plate Filtration, filtered liquid carried out the film under vacuum vaporizer concentrate, the concentrated solution volume is 1/10 of a filtered liquid volume, and concentrated solution is with the wet solid of filtration and add carrier (carrier: concentrated solution=1:1, per 1 liter of concentrated solution adds 1 kilogram of carrier), after the mixing, carry out heat-wind circulate drying, in three-dimensional meal mixer, mix powder after being crushed to the 60-80 order, and pack.
Embodiment 2: the acute toxicity of golden mushroom and genetoxic survey report
One. experiment material
1. golden mushroom: be light brown powder, adopt the method preparation of embodiment 1.
2. animal: cleaning level ICR mouse is available from dimension tonneau China laboratory animal technology company limited, and conformity certification number is SCXK (capital) 2002-0003.Chemical reagent 2-aminofluorene (2-AF), 2,4,7. trinitro--9-Fluorenone, sodium azide (NaN3), ametycin (MMC), 1,8 dihydroxyanthraquinone, Yihong is available from Beijing chemical reagents corporation.Endoxan: SHANXI POWERDONE PHARMACEUTICAL.,LTD's product, lot number: 20050604.
Two. method
1. chmice acute toxicity test: adopt once the maximum method of limiting the quantity of, 20 of the healthy ICR mouse of body weight 18～22g cleaning level, male and female half and half.Behind the fasting 16h, tried thing, observed for 2 weeks continuously by 10g/kg body weight per os.Record toxicity symptom and death condition.
2.Ames test: select for use through identifying satisfactory mouse typhus Salmonella histidine defect type TA97, TA98, TA100, TA102 four strain experimental strains, adopt flat board to mix method, divide to add and do not add two kinds of S9 mixed solutions.After being tried the thing heat sterilization, establish 5 dosage groups altogether, be respectively 200,500,1000,2500,5000 μ g/ wares (every ware is given and tried the thing amount is 0.1ml).Establish blank, positive control (9 one Fluorenones, 2-AF, NaN3, MMC, 1.8 1 istizin) simultaneously.All make 3 parallel wares for every group.Cultivate 48h down at 37 ℃, count every ware and return the change colony number, obtain every group of average colony number.Returned and become colony number and surpass from what beam back change and can be judged to be the positive more than 2 times and when dosage one reaction relation is arranged when trying thing.
3. mouse marrow cell micro nuclear test: select 50 of body weight 26～30gICR mouse for use, be divided into 5 groups at random, every group 10 and male and female half and half, dosage is divided into 1250mg/kg, 2500mg/kg, the 5000mg/kg body weight is established solvent control group (water) and positive controls (endoxan 50mg/kg body weight) simultaneously.Each group all adopts 2 per os to be tried thing, and midfeather 24h puts to death animal in the 2nd time to 6h after being tried thing, gets a side femur and prepares bone marrow smear.Microkernel incidence in the microscopy counting polychromatic erythrocyte of Gimsa dyeing back.Adopt Poinsson distribution u-test to carry out statistical study.
4. mouse sperm deformity test: select 25 of the male sexual maturity mouse of body weight 25～29g, be divided into 5 groups at random, 5 every group, if 1250mg/kg, 2500mg/kg, 3 dosage groups of 5000mg/kg body weight are established solvent control group (water) and positive controls (endoxan 50mg/kg body weight) simultaneously.The 5d per os is tried thing, every day 1 time continuously.All, get the both sides epididymis in giving the 35th day after being tried thing to put to death animal first, preparation sperm sample, Yihong dyeing, high power lens is checked the sperm morphology of every animal down, 1000 of counting structural integrity sperms, data are carried out statistical study through X2.
Three. the result
1. acute toxicity test: in the observation period of 14d. activity and the body weight gain of animal are normal, none animal dead.It is equal to male and female its mouse oral medium lethal dose (LD50) to show that this is tried thing〉the 10g/kg body weight.
2.Ames test: the results are shown in Table 1.This tried thing to TA97, TA98, TA100, TA102 four strain test strains no matter directly effect (S9) or through the metabolism activation effect (+S9) all do not cause from beaming back parameter and increase, do not have dose-effect relationship yet, illustrate that being tried the agent amount does not have mutagenicity when 200-5000 μ g/ ware.
Table 1: golden mushroom Salmonella reversion test result
Annotate: (1) 9-Fluorenone (0.5 μ g/ ware), (2) 2-aminofluorene (20 μ g/ ware), (3) sodium azide (1.5 μ g/ ware), (4) ametycin (4.0 μ g/ ware), (5) 1,8 dihydroxyanthraquinones (50 μ g/ ware)
3. mouse marrow cell micro nuclear test: by table 2 visible golden mushroom 1250mg/kg, 2500mg/kg, three dosage groups of 5000mg/kg microkernel incidence all in normal range, learn to handle by statistics and negative control group there was no significant difference (P〉0.05) relatively, positive controls and negative control group comparing difference remarkable (P＜0.01).
Table 2: golden mushroom is to the influence (n=5) of mouse Bone marrow cells micronucleus incidence
Annotate: compare with negative control:
*P＜0.01
4. mouse sperm deformity test: as seen by table 3, the rate of teratosperm of 3 dosage groups of golden mushroom is all in normal range, learn to handle by statistics with negative control group and relatively do not have significance difference (P〉0.05), positive controls and negative control group comparing difference remarkable (P＜0.01).
Table 3: golden mushroom is to the influence of mouse sperm deformity incidence
Annotate: compare with negative control:
*P＜0.05
Four. conclusion: select once the maximum method of limiting the quantity of, Salmonella reversion test, mouse marrow cell micro nuclear test, sperm malformation test to observe the acute and genetoxic that is tried thing.The result shows that to be tried the thing golden mushroom equal to male and female its mouse oral medium lethal dose (LD50)〉the 10g/kg body weight.Press the true border of acute toxicity dose grading standard nontoxic level material.No matter add and do not add metabolism activation thing S9, golden mushroom becomes the bacterium colony number average in normal range to TA97, TA98, TA100, returning of TA102 bacterial strain, there is no dosage one reaction relation; Each dosage group micronuclear rates of golden mushroom, rate of teratosperm and normal control group relatively do not have significant difference.Point out in vivo and in vitro under the condition, golden mushroom does not all have mutagenesis.Therefore, golden mushroom is safe aspect genetoxic.
Embodiment 3: golden mushroom is to the research of piglet growth performance and feed conversion rate influence
One. test materials
1. golden mushroom: embodiment 1 method manufacturing, the polysaccharide net content is 10%.
2. olaquindox: the big new poplar veterinary drug company in Hunan, Shanghai provides, and content is 50%.
3. laboratory animal: the wean three way cross piglet (hybridized pig of Ningxiang pig and the big York of Du Luo Borneo camphor) of selecting 65 body weight close (mean body weight is 13.91kg) for use, be divided into 4 processing at random according to body weight, each handles female, male ratio unanimity, each handles 3 repetitions (wherein the microbiotic group is 2 repetitions), and each repeats 6 pigs.Group is:
(1) the golden mushroom daily ration (3,4, No. 8 hurdles) of 250ppm is added in processing 1
(2) the golden mushroom daily ration (2,6, No. 9 hurdles) of 500ppm is added in processing 2
(3) handle 3 and add microbiotic olaquindox daily ration (10, No. 11 hurdles)
(4) handle 4 control group daily rations (control group, 1,5, No. 7 hurdle).
Two. test method
1. feeding and management
Measure the conventional nutritive ingredient of all feeds raw material before the test earlier, according to nutritional needs and the physilogical characteristics of weanling pig, and with reference to NRC (1998) nutritional need recommended amounts standard preparation daily ration.The control group pig is fed and is not added the basal diet of plain mushroom polysaccharide or medicine, raises by the weanling pig Managed Solution, and each feed addition is freely drunk water with clout degree of being slightly in the pit of having enough.Before on-test and duration of test, by the epidemic prevention of pig farm normal procedure, expelling parasite.Test period is 30 days.
Table 4: test with basal diet and trophic level
2. growth performance index and mensuration
The body weight of every pig is respectively organized in weighing respectively at on-test, when finishing, and (fracture 12 hours) carries out under the state of weighing in the morning on an empty stomach, calculates average daily gain of each group.Every multiple food consumption managed everywhere in record after on-test, calculates average daily ingestion amount of full phase (ADFI), average daily gain (ADG) and material anharmonic ratio (F/G).
Three. result and analysis
1. the variation of growth performance index
Table 5: the various dose golden mushroom is to the influence of weanling pig growth performance
| The plain mushroom polysaccharide of 250 gram/tons, not added with antibiotic | The plain mushroom polysaccharide of 500 gram/tons, not added with antibiotic | Do not add plain mushroom polysaccharide, added with antibiotic (olaquindox) | Do not add plain mushroom polysaccharide, not added with antibiotic | |
| First starting weight Initial Weight (Kg) | 14.13 | 15.53 | 12.10 | 14.90 |
| The heavy Finish weight (Kg) in end | 34.47 | 34.50 | 33.20 | 32.00 |
| Net gain (Kg) | 20.33 | 18.97 | 19.9 | 18.3 |
| Average daily ingestion amount ADFI (g) | 1133 | 1100 | 1100 | 1000 |
| Average daily gain ADG (g) | 450 | 420 | 440 | 410 |
| Material anharmonic ratio F/G | 2.49 | 2.58 | 2.30 | 2.64 |
By table 5 as seen, net gain, ADFI and the ADG that adds 250 gram/ton polysaccharide groups is greater than the added with antibiotic group with do not add not added with antibiotic group of polysaccharide; The net gain, ADFI and the ADG that add 500 gram/ton polysaccharide groups are greater than not adding not added with antibiotic group of polysaccharide; The F/G that adds 250 gram/ton polysaccharide groups and add 500 gram/ton polysaccharide groups is less than adding not added with antibiotic group of polysaccharide, but greater than the added with antibiotic group.Simultaneously, the effect of plain mushroom polysaccharide low dosage is better than high dosage, has reached the growth performance of microbiotic olaquindox group, so 250ppm and 500ppm Piao mushroom polysaccharide addition does not have significant difference.
Four. conclusion
This result of study shows, adds the plain mushroom polysaccharide of 250 gram/tons group and can increase the weanling pig day weight gain, improves food conversion ratio, and reduces feed cost.
Embodiment 4: golden mushroom is to the influence of weanling pig periphery blood T lymphocyte activity of conversion
One. material
1. golden mushroom: with embodiment 3.
2. microbiotic olaquindox: with embodiment 3.
3. laboratory animal: select the wean three way cross piglet (Du Luoke * length white * big York) of 72 body weight close (mean body weight is 14.2kg) for use, be divided into 4 experimental group at random, each handles female, male ratio unanimity, and each handles 3 repetitions, and each repeats 6 pigs.Handle 1 for adding the golden mushroom daily ration of 250ppm, handle 2 for adding the golden mushroom daily ration of 500ppm, handle 3 for adding microbiotic olaquindox daily ration, handling 4 is basal diet.Measure the conventional nutritive ingredient of all feeds raw material before the test earlier, according to nutritional needs and the physilogical characteristics of weanling pig, and with reference to recommendation of NRC (1998) nutritional need and feeding standard preparation daily ration.Raise by the weanling pig Managed Solution, each feed addition is freely drunk water with clout degree of being slightly in the pit of having enough.
Two. method:
1. sample collecting
Respectively at after on-test the 14th, 28 and 42d handle 6 examinations of picked at random pig from each, aseptic collection precaval vein blood sample, anticoagulant heparin is used to measure LTA.
2. the periphery blood T lymphocyte activity of conversion is measured
Adopt mtt assay.With 1 times of anti-freezing blood sample Hanks ' liquid dilution, carefully be added in the lymphocyte separation medium upper strata, 3000rmin
-1Centrifugal 20min, cloud buffy coat in the middle of drawing is washed 2 times 1500rmin with serum-free RPMI1640 nutritive medium
-1Centrifugal 15min.Behind the viable count, adjusting cell concn with the RPMI1640 nutrient solution is 2.5 * 10
6Individual mL-1 joins in the 96 porocyte culture plates, and 80 μ l/ holes, every hole add 20 μ L ConA (80 μ gmL again
-1), each sample repeats 4 holes, places 37 ℃, 5%CO2 incubator to cultivate 44h then, and every hole adds MTT20 μ L, and (the PBS liquid with pH7.4 is made into 5mgmL
-1Solution), after continue cultivating 4h, every hole adds lysate DMSO (analytical pure) 100 μ L, cell plate is placed concussion 5min dissolves precipitation fully on the microwell plate oscillator, on the enzyme linked immunological instrument, detect extinction (OD570) value at 570nm place, as the index of expression LTA.
Calculate 4 hole mean value and standard deviations, analyze, relatively the lymphocyte transformation situation with the SPSS software statistics.Test group OD value is significantly higher than control group, shows that Chinese medicine significantly promotes lymphocyte transformation; Otherwise then show remarkable inhibition lymphocyte transformation.
Three. result and analysis
Table 6: the various dose golden mushroom is to the influence (OD570 of weanling pig periphery blood T lymphocyte activity of conversion
Annotate: mark different alphabetical persons with line data and represent significant difference (P＜0.05).
By table 6 as seen, when D14 and D28, the OD570 of 250ppm and 500ppm golden mushroom group all is higher than olaquindox group and basal diet group (P〉0.05); During D42, the OD570 of 250ppm golden mushroom group is significantly higher than olaquindox group (P＜0.05), is higher than 500ppm golden mushroom group (P〉0.05), and the OD570 of 500ppm golden mushroom group also is higher than olaquindox group and basal diet group (P〉0.05).Above-mentioned test-results shows that golden mushroom can strengthen the cellular immune function of weanling pig, and the low dosage anaphase effect is more obvious.
Four. conclusion
This result of study shows that the low dosage golden mushroom can strengthen weanling pig immunizing power and disease resistance by increasing the peripheral blood lymphocyte activity of conversion, thereby reduces the harm of ablactation stress to body.
Embodiment 5: golden mushroom is to the influence of immunoglobulin (Ig) in the weanling pig serum
One. material
1. golden mushroom: with embodiment 3.
2. microbiotic olaquindox: with embodiment 3.
3. laboratory animal: the wean three way cross piglet (Du Luoke * length white * big York) of selecting 72 body weight close (mean body weight is 14.2kg) for use, be divided into 4 processing at random according to body weight, each handles female, male ratio unanimity, and each handles 3 repetitions, and each repeats 6 pigs.Handle 1 for adding the golden mushroom daily ration of 250ppm, handle 2 for adding the golden mushroom daily ration of 500ppm, handle 3 for adding microbiotic olaquindox daily ration, handling 4 is basal diet.Measure the conventional nutritive ingredient of all feeds raw material before the test earlier, according to nutritional needs and the physilogical characteristics of weanling pig, and with reference to recommendation of NRC (1998) nutritional need and feeding standard preparation daily ration.Raise by the weanling pig Managed Solution, each feed addition is freely drunk water with clout degree of being slightly in the pit of having enough.
Two. method:
Respectively at after on-test the 14th, 28 and 42d handle 6 examinations of picked at random pig, precaval vein blood sampling, 3000rmin from each
-1Centrifugal 10min separation of serum is used for the mensuration of immunoglobulin (Ig).
2. immunoglobulin content is measured in the blood plasma
Utilize CX4 type fully automatic blood Biochemical Analyzer (Beckman company product),, measure the content of immunoglobulin IgG, IgM and IgA (Beijing Leadman Biochemistry Technology Co., Ltd.'s product) in the blood plasma according to the related kit explanation; Utilizing the ELISA method to measure hog cholera antibody tires.
Three. result and analysis
1.D14 the time various dose golden mushroom to the weanling pig serum immune globulin content or the influence of tiring
By table 7 as seen, during D14, the IgG content of 250ppm golden mushroom group is higher than 250ppm golden mushroom group and basal diet group (P〉0.05), be significantly higher than olaquindox group (P＜0.05), the IgG content of 500ppm golden mushroom group also is higher than olaquindox group and basal diet group (P〉0.05); The IgA content of 250ppm and 500ppm golden mushroom group all is higher than basal diet group (P〉0.05); The IgM content of 250ppm golden mushroom group be higher than other each handle (P〉0.05); The hog cholera antibody of 250ppm golden mushroom group is tired and is higher than 500ppm golden mushroom group and olaquindox group (P〉0.05).
Table during 7:D14 the various dose golden mushroom to the weanling pig serum immune globulin content or the influence of tiring
Annotate: after serum sample 1:20 doubly dilutes, adopt ELISA kit measurement hog cholera antibody to tire, measurement result is with the positive numerical value of the sample ratio value representation divided by negative control, is higher than 2.1 just positively, and the high more expression protectiveness of ratio is good more.
2.D28 the time various dose golden mushroom to the weanling pig serum immune globulin content or the shadow of tiring
By table 8 as seen, during D28, the IgG of 250ppm and 500ppm golden mushroom group and IgM content all are higher than olaquindox group and basal diet group (P〉0.05); The IgA content of 250ppm and 500ppm golden mushroom group all is higher than olaquindox group and basal diet group (P〉0.05); The hog cholera antibody of 250ppm golden mushroom group is tired and is higher than 500ppm golden mushroom group and basal diet group (P〉0.05), is significantly higher than olaquindox group (P＜0.05).
Table 8:D28 various dose golden mushroom is to the weanling pig serum immune globulin content or the influence of tiring
3.D42 the time various dose golden mushroom to the weanling pig serum immune globulin content or the influence of tiring
By table 9 as seen, during D42, the IgG content of 250ppm golden mushroom group be higher than other each handle (P〉0.05), the IgG content of 500ppm golden mushroom group is higher than olaquindox group (P〉0.05); The IgA of 250ppm and 500ppm golden mushroom group and IgM content all are higher than olaquindox group and basal diet group (P〉0.05); The hog cholera antibody of 250ppm and 500ppm golden mushroom group tire and all be significantly higher than olaquindox group (P＜0.05), be higher than the basal diet group (P〉0.05).
Table 9:D42 various dose golden mushroom is to the weanling pig serum immune globulin content or the influence of tiring
Four. conclusion
This result of study shows that golden mushroom can strengthen weanling pig immunizing power and disease resistance by increasing the approach such as generation of piglet immunological sphaeroprotein, thereby reduces the harm of ablactation stress to body, and the middle and later periods effect obviously, and low dosage is better than high dosage.
Embodiment 6: plain mushroom polysaccharide is to the influence of meat chicken production performance
One. experimental animal and test design
This test selects for use 1 age in days to like to dial (AA) public young 540 increasingly.Test is divided into 5 processing, and each handles 6 repetitions, and each repeats 18 chickens.Handling 1 is the negative control group of added with antibiotic not, handling 2 is the positive control group of added with antibiotic, handle 3,4,5 test group of adding plain mushroom glycan feed additive for added with antibiotic not, three stages of fryer Piao mushroom polysaccharide addition is respectively 100ppm, 200ppm, 300ppm.The trophic level of each treatment group meets 2004 editions " fowl raising standard " of China.Test design sees Table 10, table 11.
Table 10: test design (1)
Table 11: test design (2)
Two. method
Test is carried out at China Agricultural University's poultry respiratory chamber, and test chicken adopts three layers and raises in cages.Feed free choice feeding and drinking-water to dry mash.Temperature is controlled between 20～26 ℃, relative humidity 45%～55%.According to normal immune programme for children immunity, observe chicken group health condition and mental status every day.
1. test daily ration
Test daily ration and trophic level see Table 12.Test each treatment group trophic level and be homenergic, etc. albumen.
2. detection index
Weigh and write down feed consumption rate at 1 age in days, 21 ages in days, 42 ages in days, 49 ages in days respectively, calculate daily ingestion amount (ADFI), day weight gain (ADG), feed efficiency (FCR), the chicken number of total chicken number and 21 ages in days, 42 ages in days, 49 ages in days when record advances chicken calculates mortality ratio.
3. statistical study
Testing data compares with Duncan with One-way ANOVA routine processes in the SPSS12.0 software.
Three. result and discussion
Test-results sees Table 13.
1. food consumption
From table 13, learn, 1～21 age in days and 1～49 age in days each handle between daily ingestion amount difference not significantly (P〉0.05); 22～42 ages in days processing 3 and processing 4 significant differences (P＜0.05), and it is minimum to handle 3 groups of food consumptions; Handle during 43～49 ages in days 4 groups food consumption with handle 1 group and handle 2 groups of differences extremely significantly (P＜0.01), handle 3 groups and handle 5 groups the daily ingestion amount utmost points and be markedly inferior to and handle 1 group (P＜0.01), and it is minimum to handle 4 groups daily ingestion amount in each treatment group.
2. day weight gain
From table 13, learn day weight gain difference not significantly (P〉0.05) between 0～21 age in days and each treatment group of 0～49 age in days; 22～42 ages in days are handled 4 groups the day weight gain utmost point and are higher than significantly and handle 1, handle 3 and handle 5 groups (P＜0.01), handle 5 groups the day weight gain utmost point and be markedly inferior to and handle 2 and handle 4 groups (P＜0.01), and it is the highest in the reason group throughout to handle 4 groups day weight gain; Handle 4 groups day weight gain during 43～49 ages in days and be significantly higher than and handle 2 groups (P＜0.05), and it is the highest in the reason group throughout to handle 4 groups day weight gain.
3. feed efficiency
From table 13, learn feed efficiency difference not significantly (P〉0.05) between each treatment group when 0～21 age in days and 0～49 age in days; Handle during 22～42 ages in days 2 and handle 4 groups feed efficiency with handle 1 and handle 5 groups of differences extremely significantly (P＜0.01), the feed efficiency of wherein handling 4 groups is the highest in the reason group throughout.Handle 4 feed efficiency during 43～49 ages in days and handle 2 groups of differences extremely significantly (P＜0.01), the feed efficiency of handling 4 groups is the highest in the reason group throughout.
4. mortality ratio
The death condition of chicken shows in the process of the test, and the equal difference of chicken mortality ratio of 1～21 age in days, 22～42 ages in days, 43～49 ages in days and 1～45 age in days is not remarkable.
Four. conclusion
This test-results shows that the production performance of handling 4 groups is best, and is not remarkable with positive control group (handling 2 groups) difference, and secondly for handling 3 groups, the poorest is negative control group (handling 1 group) and handles 5 groups.Show that interpolation 200ppm Piao mushroom polysaccharide effect is better in the daily ration of broiler.
This test is found, the effect of glycan feed additive in fryer show as the mid-term of test and later stage significantly better than test early stage, this may be since the pre-stage test chicken food consumption of test and the addition of polysaccharide all less due to, along with the increase of test chicken food consumption and glycan feed additive scale of feeding, storage effect occurred and caused production performance to be clearly better.
Claims (6)
1. golden mushroom is characterized in that: adopt following method preparation: as producing bacterial classification, adopt following fermentative medium formula with needle mushroom Flammulina velutipes (Fr.) Sing.As5.632, carry out liquid submerged fermentation:
Every liter of culture medium prescription is as follows: wheat bran 20g, corn 5g, sucrose 10g, bean powder 50g, potassium primary phosphate 1g, sal epsom 0.5g add water and are settled to 1 liter;
Inoculum size is 5-10%, and culture volume accounts for the 60-70% of fermentor tank volume, and leavening temperature is 22-25 ℃, and air flow is 1: 0.04-1.0, amplify step by step through shaking bottle, liquid seeds jar, liquid fermentation tank, and every grade of fermentation time is 72-120 hour;
Fermentation secondary fermentation liquid after filtration, filtered liquid is concentrated, concentrated solution with filter wet mycelium and carry out drying, pulverize.
2. product that contains golden mushroom is characterized in that: adopt following method preparation: as producing bacterial classification, adopt following fermentative medium formula with needle mushroom Flammulina velutipes (Fr.) Sing.As5.632, carry out liquid submerged fermentation:
Every liter of culture medium prescription is as follows: wheat bran 20g, corn 5g, sucrose 10g, bean powder 50g, potassium primary phosphate 1g, sal epsom 0.5g add water and are settled to 1 liter;
Inoculum size is 5-10%, and culture volume accounts for the 60-70% of fermentor tank volume, and leavening temperature is 22-25 ℃, and air flow is 1: 0.04-1.0, amplify step by step through shaking bottle, liquid seeds jar, liquid fermentation tank, and every grade of fermentation time is 72-120 hour;
Fermentation secondary fermentation liquid after filtration, filtered liquid concentrated after, in concentrated solution and filtration wet mycelium, add carrier, carry out drying, mix powder after the pulverizing.
3. a kind of product that contains golden mushroom according to claim 2 is characterized in that: described carrier is the corn cob fine powder, and the 80-100 order adds by 1: 1 with concentrated solution.
4. make the golden mushroom liquid fermentation producing technology for one kind, it is characterized in that: as producing bacterial classification, adopt following fermentative medium formula with needle mushroom Flammulina velutipes (Fr.) Sing.As5.632, carry out liquid submerged fermentation:
Every liter of culture medium prescription is as follows: wheat bran 20g, corn 5g, sucrose 10g, bean powder 50g, potassium primary phosphate 1g, sal epsom 0.5g without adjust pH, add water and are settled to 1 liter;
Inoculum size is 5-10%, and culture volume accounts for the 60-70% of fermentor tank volume, and leavening temperature is 22-25 ℃, and air flow is 1: 0.04-1.0, amplify step by step through shaking bottle, liquid seeds jar, liquid fermentation tank, and every grade of fermentation time is 72-120 hour; Fermentation secondary fermentation liquid after filtration, filtered liquid is concentrated adds carrier to concentrated solution with filtering in the wet mycelium, carries out drying, pulverizes.
5. a kind of manufacturing golden mushroom liquid fermentation producing technology according to claim 4 is characterized in that: being filtered into through Plate Filtration of described fermentation secondary fermentation liquid.
6. a kind of manufacturing golden mushroom liquid fermentation producing technology according to claim 4 is characterized in that: described filtered liquid is concentrated, for the thin film concentration method with 1/8 to 1/10 of filtered liquid simmer down to original volume.
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| CN107488597A (en) * | 2017-09-01 | 2017-12-19 | 山东省科创食用菌产业技术研究院 | A kind of method of liquid state fermentation production Needle mushroom strain |
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| CN1513879A (en) * | 2003-08-14 | 2004-07-21 | 中国药科大学 | Germanium enriced gold needle mushroom polysaccharide and its making metehod and use |
| CN1711885A (en) * | 2004-06-24 | 2005-12-28 | 王冬 | Non-pollution nuisance composite mycelium active forage and production thereof |
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| CN1513879A (en) * | 2003-08-14 | 2004-07-21 | 中国药科大学 | Germanium enriced gold needle mushroom polysaccharide and its making metehod and use |
| CN1711885A (en) * | 2004-06-24 | 2005-12-28 | 王冬 | Non-pollution nuisance composite mycelium active forage and production thereof |
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