Background technology
Glossy ganoderma (Ganoderma lucidum (Leyss ex Fr.) Karst.) is the traditional rare medicinal fungi of China, belongs to Basidiomycetes.For thousands of years, glossy ganoderma is owing to its good curative effect is described as " celestial grass ".The modern science and technology result of study shows that the main active of glossy ganoderma is GL-B and ganoderic acid.GL-B is that a big class is with β-(1,3) and β-(1,4) polysaccharide or the heteropolysaccharide macromolecular substances that are formed by connecting of key, it is a kind of important immunopotentiator, can significantly activate the autoimmunity system, simultaneously it also has antibiotic, antiviral, anti parasitic, antitumor, radioresistance and improves a series of biologically actives such as breeding performonce fo animals.Ganoderic acid is a class triterpene compound, studies show that ganoderic acid can suppress the growth of 7 kinds of pathogenic bacteria (as staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella Pneumoniae, shigella dysenteriae, bacillus ceylonensis A, salmonella typhi), keeps the health of animal intestinal; It and GL-B use simultaneously, and GL-B is had synergistic effect.
Main at present liquid fermentation and the solid state cultivation dual mode of adopting of the production of glossy ganoderma.Adopt the liquid fermentation mode, during the fermentation, a part of polysaccharide and ganoderic acid are directly secreted in zymotic fluid, form the outer ganoderic acid of exocellular polysaccharide and born of the same parents; Then exist ganoderic acid in intracellular polyse and the born of the same parents in the ganoderma lucidum mycelium, need carry out earlier just further extracting after the broken wall treatment.Studies show that born of the same parents' ganoderma active material interior, that born of the same parents are outer that the ganoderma lucidum liquid fermentation obtains all has similar activity.Adopt the solid state cultivation mode, its polysaccharide and ganoderic acid composition all are present in the born of the same parents, need carry out earlier equally just further extracting after the broken wall treatment.
Feed addictive improves animal immunizing power, tachyauxesis in the livestock and poultry cultivation process, improve food conversion ratio, reduce aspects such as feeding cost plays a part indispensable.Along with the raising of people's food security consciousness and the enhancing of health perception, and the further quickening of the international market paces of animal products, develop nuisanceless animal husbandry, produce the green livestock and poultry product, the market competitiveness that strengthens China's animal products has become the top priority of China's herding industry.The green feed additive of research and development Effictive nuisancelless then becomes the task of top priority of this research field.
That green feed additive is meant in broad terms is pollution-free, noresidue, anti-disease, somatotrophic natural additive, sets about the research and development of being correlated with abroad before more than 30 years, and domestic just starting in recent years.At present, the scientific worker has has researched and developed green feed additives such as Chinese herbal and crude drugs preparations, enzyme preparation, microorganism formulation, acidulant, mould inhibitor, prebiotics or chemical probiotic.But these additives all exist following one or more weak points, as poor heat stability, and easy inactivation, adding proportion is bigger than normal, cost height, resource-constrained etc.
Because that the contained active material of glossy ganoderma has is efficient, nontoxic, improve characteristics such as immunity, anti-bacteria and anti-virus, noresidue, meets the requirement of green feed and green cultivation, is a kind of brand-new natural green feed addictive.At present, the existing report for preparing feed addictive on a small quantity about glossy ganoderma, there is the scholar that the ganoderma lucidum fruitbody of solid state cultivation is pulverized the preparation feed addictive or/and mycelium is directly dried, also have the scholar with the ganoderma lucidum fruitbody of solid state cultivation or/and mycelium by further extracting GL-B, prepares feed addictive with GL-B.It is long that but the mode that adopts solid state cultivation exists biocycle, thereby owing to the inhomogeneity of culture medium causes the content fluctuation of its bioactive ingredients to influence shortcomings such as feed addictive quality; And as ganoderma lucidum fruitbody is pulverized the preparation feed addictive or/and mycelium is directly dried, its active ingredient GL-B and ganoderic acid still in born of the same parents, are difficult to fully be assimilated by animal, fall flat; And further prepare feed addictive from ganoderma lucidum fruitbody or/and obtain GL-B the mycelium, and needing to extract, concentrate, more complicated extraction step such as alcohol precipitation, Separation of Solid and Liquid, production cost improves greatly.
And patent ZL200410042719.8 provides a kind of preparation method of Polysaccharide-Peptide complex immunologic facilitation feed addictive, its main component is that glossy ganoderma is or/and the zymotic fluid of rainbow conk, through the feed addictive that obtains four kinds of formulations after further the back processes respectively is crystal refining product, absorption pulvis, liquor, mycelium pulvis, wherein first three kind is to prepare by liquid fermentation liquid, and the 4th kind is that mode by the liquid spawn solid state cultivation obtains behind the mycelium further that processing and preparing obtains.This patent has overcome the deficiency that ganoderma lucidum fruitbody that the solid state cultivation mode obtains or mycelium prepare feed addictive to a certain extent.But this method needs more step (as preparation crystal refining product, absorption pulvis process) in the processing of ganoderma lucidum fermented liquid, perhaps can not utilize active ingredient all in the ganoderma lucidum fermented liquid (as preparation crystal refining product and liquor process) cmpletely.
Summary of the invention
The objective of the invention is to some shortcomings, a kind of cost is low, step is few, the ganoderma lucidum fermented liquid effective ingredient is utilized cmpletely the feed that contains active components of glossy ganoderma and the method for feed addictive are provided at above-mentioned green feed additive.
Realization the objective of the invention is to realize by following steps:
Step 1: the Separation of Solid and Liquid of ganoderma lucidum fermented liquid
Adopt the liquid fermentation mode,, obtain glossy ganoderma fermentation supernatant and glossy ganoderma bacterium slag ganoderma lucidum fermented liquid centrifugation under 5000 rpms condition of fermentation ends.
Step 2: contain the preparation of active components of glossy ganoderma feed
Adopt the back spraying coating process that the glossy ganoderma fermentation supernatant that step 1 obtained is mixed with feed, preparation contains the feed of active components of glossy ganoderma, and adding proportion is added 0.5~1.5 milliliter of glossy ganoderma fermentation supernatant for the per kilogram feed.Described feed is mainly the poultry feed, and described back spraying coating process can adopt air spraying technology, and its operating condition is atomisation pressure: 0.2~0.5MPa, 20~40 ℃ of temperature of charge, charging rate: 10mL/s; Also can adopt airless spraying technology, its operating condition is vacuum: 0.02~0.04MPa, rushes down pressure time: 80~120s, fluid temperature: 30~50 ℃.
Step 3: the preparation of glossy ganoderma bacterium slag secondary fermentation contains the feed addictive of active components of glossy ganoderma
The culture medium of the glossy ganoderma bacterium slag that step 1 obtained being formed secondary fermentation for the basis, insert lactobacillus inoculation, inoculum concentration is 5%~8% (volume mass than), 30~40 ℃ of condition bottom fermentations 24~48 hours, can carry out drying by this area conventional method after the fermentation ends.
Secondary fermentation culture medium of the present invention consist of glossy ganoderma bacterium slag, corn flour, wheat bran, wherein corn flour and wheat bran adding proportion are 1: 1, after glossy ganoderma bacterium slag and a certain amount of corn flour, wheat bran mixed, make that institute's water content is controlled at 60%~65% (mass percent) in the secondary fermentation culture medium; Culture medium is heated to 100 ℃, and keeps 40min; Be cooled to 60 ℃, the amount of adding 100 international units by every gram corn flour in the culture medium adds carbohydrase and mixes in culture medium; Continue to be cooled to insert in the lactic acid bacteria below 40 ℃ and ferment.
Lactobacillus inoculation of the present invention for example can be the CGMCC No.1.1854 available from Chinese common micro-organisms culture presevation administrative center (CGMCC) for lactobacillus acidophilus commonly used.This area technical method commonly used is adopted in the preparation of lactic acid seed.
The preparation of ganoderma lucidum fermented liquid of the present invention can be adopted this area preparation method commonly used.As the glossy ganoderma bacterium can be CGMCC No.5.535 available from Chinese common micro-organisms culture presevation administrative center (CGMCC); The preparation of ganoderma lucidum fermented liquid can adopt following process: a, inclined-plane to cultivate: lucidum strain is inoculated on the slant medium, is cultured to mycelium and covers with the inclined-plane under 25 ℃.The concrete preparation method of described slant medium is: will cut shape or little bulk behind the peeling potatoes into pieces, take by weighing in 200 gram addings, 500 ml waters and boil after 40 minutes with 4 layers of filtered through gauze, in filtered juice, add 20 gram glucose, add water then and be settled to 1000 milliliters, add 20 gram agar and heating again, treat that agar dissolves the back packing fully, sterilization, standby.B, seed culture: good bacterial classification is grown on the inclined-plane inserted in the seed culture medium, 150 rpms of shaking speed were cultivated 7 days for 25 ℃.Described seed culture medium is (unit is every liter of gram): corn flour 20, and sucrose 20, wheat bran 15, potassium dihydrogen phosphate 0.75, epsom salt 0.5, initial pH are nature pH.C, liquid fermentation and culture: the liquid seeds that obtains transferred by the inoculum concentration of 10% (percent by volume) be equipped with in the fermentation tank of fermentation medium, 28 ℃ of cultivations, fermentation tank pressure 0.05Mpa, 120 rpms of stir speed (S.S.)s, ventilation 1: 0.5v/v/m cultivated 3 days.Described fermentation medium is (unit is every liter of gram): corn flour 22, and sucrose 18, wheat bran 10, potassium dihydrogen phosphate 1.5, magnesium sulfate 1.0, initial pH are nature pH.
The active components of glossy ganoderma feed that contains provided by the present invention mainly can be used as the use of poultry feed, uses this feed can reach the speed of growth that promotes animal, increases immunity and resistance against diseases, reduces the effect of feedstuff-meat ratio.
Provided by the present inventionly contain the additive that the active components of glossy ganoderma feed addictive mainly can be used as the poultry feed and use, press dry weight basis, the per kilogram feed adds this feed addictive in the ratio of 0.5 ‰~2 ‰ (mass percents), can promote the speed of growth of animal, stimulating immune system is grown, increase immunity and resistance against diseases, reduce the effect of feedstuff-meat ratio.
The invention has the advantages that:
1) feed and the feed addictive that contains active components of glossy ganoderma provided by the invention contains abundant GL-B and ganoderic acid, has balance immune system, antibiotic, antiviral, anti parasitic, antitumor, radioresistance etc. be physiologically active widely, can improve the immune disease-resistance ability greatly, keep the health of animal intestinal, improve production performance and the market efficiency of animal.
2) the present invention adopts the back spraying coating process that ganoderma lucidum fermented liquid is mixed with feed, can increase the uniformity that feed mixes greatly, significantly reduces because of expanded, expansion, granulation supervisor cause loss of active ingredients, fully guarantees the quality of feed.
3) the present invention carries out secondary fermentation with glossy ganoderma bacterium slag, its benefit is by the processing of " 100 ℃; keep 40min " and follow-up lactic acid bacteria fermentation process, make the ganoderma lucidum mycelium self-dissolving, GL-B and ganoderic acid in the born of the same parents fully are released in the feed addictive, and the total losses of active components of glossy ganoderma can well fully be assimilated by animal only below 3% in the secondary fermentation process;
4) simultaneously, in the present invention glossy ganoderma bacterium slag is carried out in the process of secondary fermentation, make the content that increases lactic acid and peptide class in the feed addictive by lactobacillus-fermented, lactic acid bacterium number reaches (by dry) more than 200,000,000 in every gram feed addictive, make it contain a large amount of probios on basis of active components of glossy ganoderma again possessing, thereby improved the overall efficacy of this additive greatly.
5) method provided by the present invention all makes full use of ganoderma lucidum fermented liquid, and no waste water and dregs is got rid of, and step is simple, with low cost, is easy to extensive industrialization.
The specific embodiment
In described below all specific embodiment, the mensuration of GL-B adopts phenolsulfuric acid to measure, (Zhang Weijie writes this method in " saccharide complex Biochemical Research technology (second edition) ", publishing house of Zhejiang University, 1999) and other numerous works and both at home and abroad in the academic journal paper detailed statement is arranged all, here no longer describe in detail.The assay method of ganoderic acid according to described determined by ultraviolet spectrophotometry such as clock Kien Giang (Effect of initial pH on production of ganoderic acid and polysaccharide bysubmerged fermentation of Ganoderma lucidum.Process Biochemistry.2002,37:769-774.).
In the following specific embodiment, the water content of raw materials used corn flour and wheat bran is 12% (mass percent); Carbohydrase is provided by Wuxi Saden Bioisystech Co., Ltd, the vigor of carbohydrase is every milliliter of 100,000 international units, with pure water it is diluted to the level of every milliliter of 1000 international units before use, preserve under 4 ℃ of conditions and use, its purpose is to make carbohydrase and culture medium material fully to mix.According to fermentating culturing process of the present invention, can be in conjunction with the ABC of this area further according to producing needs expansion fermentation scale.In the process that enlarges the fermentation scale, can determine according to actual needs carbohydrase directly to be used or dilute the back and use.
The centrifugation of preparation of embodiment 1 ganoderma lucidum fermented liquid and zymotic fluid
(1) inclined-plane is cultivated: lucidum strain CGMCC No.5.535 is inoculated on the slant medium, under 25 ℃, be cultured to mycelium and cover with the inclined-plane, the concrete preparation method of described slant medium is: will cut shape or little bulk behind the peeling potatoes into pieces, take by weighing in 200 gram addings, 500 ml waters and boil after 40 minutes with 4 layers of filtered through gauze, in filtered juice, add 20 gram glucose, add water then and be settled to 1000 milliliters, add 20 gram agar and heating again, treat that agar dissolves the back packing fully, sterilization, standby.
(2) seed culture: good bacterial classification is grown on the inclined-plane inserted in the seed culture medium, the volume of seed culture medium is 100 milliliters, in 500 milliliters the triangular flask of packing into, and totally 21 bottles.Seed culture medium is (unit is every liter of gram): corn flour 20, and sucrose 20, wheat bran 15, potassium dihydrogen phosphate 0.75, epsom salt 0.5, initial pH are nature pH; Condition of culture is: 150 rpms of shaking speed, cultivated 7 days for 25 ℃.
(3) liquid fermentation and culture: cultured seed is inserted in 30 liters of fermentation tanks, and inoculum concentration is 2.1 liters, 21 liters of zymotic fluid cumulative volumes.Fermentation medium is (unit is every liter of gram): corn flour 22, and sucrose 18, wheat bran 10, potassium dihydrogen phosphate 1.5, magnesium sulfate 1.0, initial pH are nature pH.28 ℃ of cultivations, fermentation tank pressure 0.05Mpa, 120 rpms of stir speed (S.S.)s, ventilation are 10.5 liters of per minutes, cultivate 3 days.Finally obtain 20 liters of zymotic fluids.
(4) with ganoderma lucidum fermented liquid centrifugation under 5000 rpms condition, finally obtain 16.5 liters of glossy ganoderma fermentation supernatants, wherein contain GL-B 3.0 grams, 352.22 milligrams of ganoderic acids in every liter of fermented supernatant fluid; Obtain 3.6 kilograms of glossy ganoderma bacterium slags, water content 77.2%, wherein (by dry weight basis) contains GL-B 50.4 grams in the per kilogram glossy ganoderma bacterium slag, ganoderic acid 13.66 grams.
Embodiment 2 contains the preparation of ganoderma active composition feed
In the ratio that the per kilogram feed adds 1.0 milliliters of glossy ganoderma fermentation supernatants, adopt air spraying technology that the glossy ganoderma fermentation supernatant is sprayed at the feed granules surface, its operating condition is atomisation pressure: 0.2MPa, 20 ℃ of temperature of charge, charging rate: 10mL/s.The spraying index: feed mixture homogeneity 0.06%, pellet hardness reduces by 5%, active components of glossy ganoderma loss late 5.2%.Described feed is typical dregs of beans-corn type chicken feed, its nutritional labeling (mass percent): thick protein 19%, crude fat 3.0%, crude fibre 6.0%, coarse ash 8.0%, calcium 1.20%, total phosphorus 0.60%, salt 0.6%; Metabolizable energy is 3.0 kilocalories of every grams.
Embodiment 3 contains the preparation of ganoderma active composition feed
In the ratio that the per kilogram feed adds 0.8 milliliter of glossy ganoderma fermentation supernatant, adopt airless spraying technology that the glossy ganoderma fermentation supernatant is sprayed at the feed granules surface, its operating condition is vacuum: 0.02MPa, rushes down pressure time: 80s, fluid temperature: 30 ℃.The spraying index: feed mixture homogeneity 0.03%, pellet hardness reduces by 3%, active components of glossy ganoderma loss late 3.8%.Described feed is typical corn-dregs of beans-assorted dregs of rice type duck feed, its nutritional labeling (mass percent): thick protein 15%, crude fat 3.0%, crude fibre 6.0%, coarse ash 8.0%, calcium 0.91%, total phosphorus 0.68%, salt 0.3%; Metabolizable energy is 2.8 kilocalories of every grams.
Embodiment 4 contains the preparation of ganoderma active composition feed
In the ratio that the per kilogram feed adds 1.5 milliliters of glossy ganoderma fermentation supernatants, adopt air spraying technology that the glossy ganoderma fermentation supernatant is sprayed at the feed granules surface, its operating condition is atomisation pressure: 0.5MPa, 40 ℃ of temperature of charge, charging rate: 10mL/s.The spraying index: feed mixture homogeneity 0.07%, pellet hardness reduces by 4%, active components of glossy ganoderma loss late 5.8%.Described feed is with embodiment 3.
Embodiment 5 contains the preparation of ganoderma active composition feed
In the ratio that the per kilogram feed adds 1.5 milliliters of glossy ganoderma fermentation supernatants, adopt airless spraying technology that ganoderma lucidum fermented liquid is sprayed at the feed granules surface, its operating condition is vacuum: 0.04MPa, rushes down pressure time: 120s, fluid temperature: 50 ℃.The spraying index: feed mixture homogeneity 0.04%, pellet hardness reduces by 3%, active components of glossy ganoderma loss late 3.5%.Described feed is with embodiment 2.
Embodiment 6 contains the preparation of ganoderma active composition feed
In the ratio that the per kilogram feed adds 0.5 milliliter of glossy ganoderma fermentation supernatant, adopt air spraying technology that the glossy ganoderma fermentation supernatant is sprayed at the feed granules surface, its operating condition is atomisation pressure: 0.35MPa, 30 ℃ of temperature of charge, charging rate: 10ml/s.Spraying index: spraying index: feed mixture homogeneity 0.06%, pellet hardness reduces by 5%, active components of glossy ganoderma loss late 4.6%.Described feed is with embodiment 2.
Embodiment 7 contains the preparation of ganoderma active composition feed
In the ratio that the per kilogram feed adds 0.5 milliliter of glossy ganoderma fermentation supernatant, adopt airless spraying technology that ganoderma lucidum fermented liquid is sprayed at the feed granules surface, its operating condition is vacuum: 0.03MPa, rushes down pressure time: 100s, fluid temperature: 40 ℃.Spraying index: spraying index: feed mixture homogeneity 0.03%, pellet hardness reduces by 4%, active components of glossy ganoderma loss late 2.9%.Described feed is with embodiment 3.
Embodiment 8 contains the preparation of ganoderma active composition feed addictive
Add 179.16 gram corn flour and 179.16 gram wheat brans in 1 kilogram of glossy ganoderma bacterium slag, and mix, the water content in the culture medium is 60% (mass percent); Culture medium is heated to 100 ℃, and keeps 40min; Be cooled to 60 ℃, add 17.9 milliliters of carbohydrase dilutions, and mix; When temperature is reduced to 30 ℃, insert 108.7 milliliters of lactobacillus inoculations, finished in 48 hours at 30 ℃ of condition bottom fermentations.
Wherein, being prepared as follows of lactobacillus solution:
(1) inclined-plane is cultivated: lactobacillus inoculation CGMCC No.1.1854 is inoculated on the slant medium, be cultured to thalline and cover with the inclined-plane under 37 ℃, described slant medium is (unit is every liter of gram): yeast extract 5, peptone 7.5, glucose 10, potassium dihydrogen phosphate 4,, Tween 80 0.5, epsom salt 1, agar powder 15, pH6.5 sterilized 20 minutes for 121 ℃.
(2) seed culture: good bacterial classification is grown on the inclined-plane inserted in the seed culture medium, the volume of seed culture medium is 100 milliliters, in 250 milliliters the triangular flask of packing into, and totally two bottles.37 ℃ of constant temperature leave standstill to be cultivated 18 hours, and seed culture medium is (unit is every liter of gram): glucose 10, peptone 5, epsom salt 0.5, potassium dihydrogen phosphate 1, sodium chloride 5,5.5,121 ℃ of sterilizations of pH 20 minutes.
Embodiment 9 contains the preparation of ganoderma active composition feed addictive
Add 115.09 gram corn flour and 115.09 gram wheat brans in 1 kilogram of glossy ganoderma bacterium slag, and mix, the water content in the culture medium is 65% (mass percent); Culture medium is heated to 100 ℃, and keeps 40min; Be cooled to 60 ℃, add 11.5 milliliters of carbohydrase dilutions, and mix; When temperature is reduced to 35 ℃, insert 61.5 milliliters of lactobacillus inoculations, 35 ℃ of condition bottom fermentations 38 hours.
Wherein, the preparation of lactobacillus solution is with embodiment 8.
Embodiment 10 contains the preparation of ganoderma active composition feed addictive
Add 135 gram corn flour and 135 gram wheat brans in 1 kilogram of glossy ganoderma bacterium slag, and mix, the water content in the culture medium is 63.3% (mass percent); Culture medium is heated to 100 ℃, and keeps 40min; Be cooled to 60 ℃, add 13.5 milliliters of carbohydrase dilutions, and mix; When temperature is reduced to 35 ℃, insert 88.9 milliliters of lactobacillus inoculations, 32 ℃ of condition bottom fermentations 42 hours.
Wherein, the preparation of lactobacillus solution is with embodiment 8.
Embodiment 11 contains the preparation of ganoderma active composition feed addictive
Add 150 gram corn flour and 150 gram wheat brans in 1 kilogram of glossy ganoderma bacterium slag, and mix, the water content in the culture medium is 62.1% (mass percent); Culture medium is heated to 100 ℃, and keeps 40min; Be cooled to 60 ℃, add 15 milliliters of carbohydrase dilutions, and mix; When temperature is reduced to 40 ℃, insert 78 milliliters of lactobacillus inoculations, 37 ℃ of condition bottom fermentations 32 hours.
Wherein, the preparation of lactobacillus solution is with embodiment 8.
Embodiment 12 contains the preparation of ganoderma active composition feed addictive
Add 120 gram corn flour and 120 gram wheat brans in 1 kilogram of glossy ganoderma bacterium slag, and mix, the water content in the culture medium is 64.6% (mass percent); Culture medium is heated to 100 ℃, and keeps 40min; Be cooled to 60 ℃, add 12 milliliters of carbohydrase dilutions, and mix; When temperature is reduced to 40 ℃, insert 99.2 milliliters of lactobacillus inoculations, 40 ℃ of condition bottom fermentations 24 hours.
Wherein, the preparation of lactobacillus solution is with embodiment 8.
Embodiment 13 contains the preparation of ganoderma active composition feed addictive
Add 165 gram corn flour and 165 gram wheat brans in 1 kilogram of glossy ganoderma bacterium slag, and mix, the water content in the culture medium is 61% (mass percent); Culture medium is heated to 100 ℃, and keeps 40min; Be cooled to 60 ℃, add 16.5 milliliters of carbohydrase dilutions, and mix; When temperature is reduced to 40 ℃, insert 99.7 milliliters of lactobacillus inoculations, 35 ℃ of condition bottom fermentations 38 hours.
Wherein, the preparation of lactobacillus solution is with embodiment 8.
Embodiment 14 feeding trials
The feed that adopts embodiment 2 to obtain carries out the fowl raising test, establishes two parallel raising groups and 1 control group (feeds utilized for not containing the feed of ganoderma lucidum fermented liquid among the embodiment 2) altogether, raises after 45 days, has than obvious effects (table specific as follows).
Embodiment 15 feeding trials
The feed addictive that embodiment 8 is obtained is added in the duck basal feed, and the duck basal feed is with typical corn-dregs of beans among the embodiment 3-assorted dregs of rice type duck feed.Establish two parallel raising groups and 1 control group (not adding this feed addictive) altogether, wherein raise the stage addition of (10 days) duckling be 0.5% to feed addictive; The addition of raising the stage (20 days) big duck is 2.0 ‰.Raise after 30 days, use this feed addictive after, have than obvious effects (table specific as follows).
Below described embodiment of the present invention in detail, can do a lot of improvement and variation obviously for a person skilled in the art and can not deviate from essence spirit of the present invention.All these changes and improvements are all within protection scope of the present invention.