CN102919542B - Multistage-controlled-release probiotics microecological preparation composite feed for Silurus meridionalis and preparation method thereof - Google Patents
Multistage-controlled-release probiotics microecological preparation composite feed for Silurus meridionalis and preparation method thereof Download PDFInfo
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Abstract
The invention discloses multistage-controlled-release probiotics microecological preparation composite feed for Silurus meridionalis, which comprises lactobacillus acidophilus, lactobacillus plantarum and streptococcus faecalis which are cultured in large scale by culture mediums; and the multistage-controlled-release probiotics microecological preparation composite feed for Silurus meridionalis is prepared by the low-temperature spray drying technology and the technology for preparing microcapsules by carbon dioxide ice freezing and vacuum drying technology. Experimental results show that the composite feed can notably promote growth of fishes such as Silurus meridionalis; and after raising juvenile fish for 45 days, compared with a control group, the quantity of the Silurus meridionalis is increased by 12.4%, the protein efficiency is improved by 32.5% and the incidence rate is reduced by 30-40%.
Description
Technical field
The present invention relates to field of feed, in particular probiotics micro-ecological formulation mixed feed of the feeding multistage controlled release of a kind of Silurus meridionalis Chen and preparation method thereof.
Background technology
Silurus meridionalis Chen, also claims Silurus Soldatovi Meridionalis fish, and this kind of fish common disease in feeding process has: white skin disease, stigmatosis, canker, hemorrhage, tail, fin rot, enteritis disease etc.In feeding process, these common diseases have greatly affected the production performance of southern catfish and the quality of fishery-ies product.On the other hand, due to discharge of industrial wastewater, sanitary sewage etc., breeding environment further worsens, and causes fish diseases to take place frequently, and current culture fishery, the main antibiotics of wide spectrum that adopts is controlled disease, like this, not only causes the germ resistance to the action of a drug to strengthen, disturb the growth and breeding of probio flora in fish intestines, cause microecological balance imbalance, also can cause secondary pollution, have a strong impact on aquaculture and the aquatic products quality of fish.
Probiotics is under micro-ecological theory instructs, and the living microbe formulation that utilizes the material of normal microorganism or accelerate growth of microorganism to make, has function supplementary, that adjust and maintain microecological balance in animal intestinal.Thereby reach the effect of disease preventing and treating, promotion health and raising production performance.It is a kind of green, environmental protection, pure biologic product, nontoxic, have no side effect, and noresidue is polluted, and does not produce resistance, and water body is not produced to secondary pollution,
Probiotics abroad due to start-up time early, in fast development and the stage of ripeness, and at home, the research of the probiotics of prebiotic function bacterium originates in the end of the year 70 of last century, at present at home or a new industry, the starting stage at an early stage of development.In city's pin product, conventional animal probio has at present: bacillus, lactobacillus, saccharomycete, streptococcus fecalis etc.The probio pulvis of domestic use as lactobacillus preparation be all single probiotics substantially, the function singleness having, can not meet the demand in market.At present, for the common disease of Silurus meridionalis Chen, can be few for the feeding multistage controlled release probio compound micro-ecological preparation of Silurus meridionalis Chen.In addition, probiotic composition in the market, majority is the trophosome cell of non-embedding, prepared by the cryodesiccated method that individual other probiotics adopts, not only cost is high, and survival rate is lower, go back simultaneously ubiquity keeping quality poor, extend the shortcoming that number of viable reduction etc. is difficult to overcome with the holding time, the application of product is restricted, to industry development, brings many negative effects.Therefore, be necessary, for aforesaid defect, the feeding multistage controlled release of the fish such as Silurus meridionalis Chen probiotics micro-ecological formulation mixed feed and preparation method thereof is made to further research and improvement.
Summary of the invention
Technical problem to be solved by this invention is probiotics micro-ecological formulation mixed feed providing for the deficiencies in the prior art the feeding multistage controlled release of a kind of Silurus meridionalis Chen and preparation method thereof.
Technical scheme of the present invention is as follows:
The probiotics micro-ecological formulation mixed feed of the feeding multistage controlled release of a kind of Silurus meridionalis Chen, probiotics micro-ecological formulation mixed feed comprises lactobacillus acidophilus (the Lactobacillus acidophilus that expands cultivation through culture medium, be purchased from Chinese industrial microorganism fungus kind preservation administrative center, numbering: CICC 6075), Lactobacillus plantarum (Lactobacillus plantarum, be purchased from Chinese industrial microorganism fungus kind preservation administrative center, numbering: CICC 6009), enterococcus faecalis (Streptococcus faecalis, be purchased from Chinese industrial microorganism fungus kind preservation administrative center, numbering: CICC23658), adopt respectively low temperature spray drying technology, dry carbon dioxide shaft ice frozen and vacuum drying technique are prepared microcapsules technology, make the probiotics micro-ecological formulation mixed feed of the feeding multistage controlled release of Silurus meridionalis Chen.
The preparation method of described probiotics micro-ecological formulation mixed feed, comprises the following steps:
Step 1, actication of culture and expansion are cultivated;
After being activated respectively, expand cultivation, the lactobacillus acidophilus bacterial classification of refrigeration, Lactobacillus plantarum bacterial classification and enterococcus faecalis obtain lactobacillus acidophilus bacteria suspension, Lactobacillus plantarum bacteria suspension and enterococcus faecalis bacteria suspension; Respectively above-mentioned lactobacillus acidophilus bacteria suspension, Lactobacillus plantarum bacteria suspension and enterococcus faecalis bacteria suspension are filtered, and by the bacteria suspension of collecting after filtering by lactobacillus acidophilus bacterium: Lactobacillus plantarum bacterium: the part by weight of enterococcus faecalis bacteria suspension=1:1:2-2:2:1 mixes to obtain compound bacteria suspension, standby;
Step 2, the preparation of quick-release bacterium powder
The compound bacteria suspension of step 1 gained is filtered, in suspension after filtration, add account for filter after 8% to 12% beta-schardinger dextrin-of compound bacteria suspension gross weight, 5% to 8% porous-starch, soybean protein isolates 5%-8%, 13% to 16% sterilized water, adopts 900 revs/min of electric mixers to stir after 20-40 minute, adopt vacuum desiccator at vacuum condition to be-0.085MPa, temperature is dry 0.5h-1h at 25 ℃, mixes, and obtains quick-release bacterium powder 1;
The preparation of step 3, middling speed release type bacterium powder
Under aseptic technique, take the porous-starch of 6g-10g, the shitosan of 6g-10g is in beaker, the sterilized water that adds 100mL, take that this 100mL sterilized water is follow-up interpolation material with reference to amount, the acetic acid that adds 0.5wt%-0.8 wt %, adopt 1200 revs/min of electric mixers to stir after 20-40 minute, shitosan and porous-starch are dissolved completely, add the compound bacteria suspension 30g-40g being prepared by step 1, at room temperature adopt again supersonic generator concussion absorption 30-50 minute, add 2 wt % sodium alginate solns to obtain emulsion, with syringe, emulsion is clamp-oned to the Cacl that the concentration of 15-25mL is 2%
2in solution, solidify 30min film forming, after filtering, by the concentration of 50-60mL, be 3.5%Cacl
2solution soaks 10min and hardens, and refilters and obtains sample, with clear water, floats surperficial Cacl
2raffinate, then wash surface of microcapsule thalline with the physiological saline of sterilizing.Adopt the dry carbon dioxide shaft ice frozen 5-8 minute of-78 ℃ again, adopt vacuum desiccator at vacuum condition to be-0.085MPa, temperature is dry 0.5h-1h at 25 ℃, obtains middling speed release type bacterium powder 2;
Step 4, the preparation of release type bacterium powder at a slow speed
Under aseptic technique, take 7g-11g zeins matter in beaker, the food-grade anhydrous ethanol that adds 100mL, middling speed release type bacterium powder 25-35g prepared by step 3 is scattered in ethanol, adopts 1200 revs/min of electric mixers to stir after 20-40 minute, through low temperature spray drying, 90 ℃～105 ℃ of inlet temperatures, 60 ℃～65 ℃ of outlet temperatures, after spraying is dry, collects and obtain release type bacterium powder 3 at a slow speed;
The probiotics micro-ecological formulation mixed feed preparation of step 5, multistage controlled release
Respectively by quick-release bacterium powder 1, middling speed release type bacterium powder 2 and at a slow speed release type bacterium powder 3 mix by 2:1:1 to 1:1:2 part by weight, get Mixed Microbes powder, then add 7% to 9% bean dregs, 5% to 7% groundnut kernel powder, 3% to 5% WPC matter, 8 to 10% soybean protein isolates, dusty yeast 3% to 5%, fish oil 1.1% to 3.5%, corn flour 8.3 % to 10.5%, amino acid 2%, premix 1.3 % to 2.8%, starch 13 % to 15%, soy meal 10% to 13%, adopt 900 revs/min of electric mixers to stir after 20-40 minute, mix, obtain the probiotics micro-ecological formulation mixed feed of multistage controlled release.
Experimental result shows, this mixed feed can obviously promote the fish growths such as Silurus meridionalis Chen, raise after juvenile fish 45d, south catfish amount increase has improved 12.4% than control group respectively, protein efficiency ratio, PER improves 32.5%, the incidence of disease reduces 30%-40%, this probiotics mixed feed can improve the metabolism of body, supplement the nutritional labeling of body, beneficial bacterium can further be bred after entering alimentary canal, produce necessary nutriment in growth of animal process, improve the conversion ratio of feed, promote the growth of Silurus meridionalis Chen, and make the meat of Silurus meridionalis Chen more delicate, taste is more delicious, there is better edibility, bring up better economic worth.And a kind of probiotics preparation method for Silurus meridionalis Chen provided by the present invention, processing step is simple, and material therefor cost is low, is suitable for suitability for industrialized production, and range of application is wide.
The specific embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
One aspect of the present invention provides the probiotics micro-ecological formulation mixed feed of the feeding multistage controlled release of a kind of Silurus meridionalis Chen, comprise through culture medium and expand lactobacillus acidophilus (the Lactobacillus acidophilus cultivating, be purchased from Chinese industrial microorganism fungus kind preservation administrative center, numbering: CICC 6075), Lactobacillus plantarum (Lactobacillus plantarum, be purchased from Chinese industrial microorganism fungus kind preservation administrative center, numbering: CICC 6009), enterococcus faecalis (Streptococcus faecalis, be purchased from Chinese industrial microorganism fungus kind preservation administrative center, numbering: CICC23658), adopt respectively low temperature spray drying technology, dry carbon dioxide shaft ice frozen and vacuum drying technique are prepared microcapsules technology, make the probiotics micro-ecological formulation mixed feed of the feeding multistage controlled release of Silurus meridionalis Chen.
First adopt low temperature spray drying technology respectively lactobacillus acidophilus, Lactobacillus plantarum, enterococcus faecalis to be made to single wall material probiotics, lactobacillus acidophilus bacterium powder, Lactobacillus plantarum bacterium powder, enterococcus faecalis bacterium powder are mixed by 3: 2: 1 part by weight to 2:2:3, then dry carbon dioxide shaft ice frozen and vacuum drying technique are made composite bacterium powder the probiotics of double wall material again; Finally, by the probiotics of the probiotics of three kinds of bacterium mixtures, solid wall material and double wall material by a certain percentage Silurus meridionalis Chen feed base thing mix, make the probiotics micro-ecological formulation mixed feed of the feeding multistage controlled release of Silurus meridionalis Chen.
The present invention provides a kind of probiotics micro-ecological formulation mixed feed preparation method for the feeding multistage controlled release of Silurus meridionalis Chen on the other hand, it is the main compound micro-ecological preparation of preparing that described method be take lactobacillus acidophilus, Lactobacillus plantarum, enterococcus faecalis, operates in accordance with the following steps:
Step 1, actication of culture and expansion are cultivated
After being activated respectively, expand cultivation, the lactobacillus acidophilus bacterial classification of refrigeration, Lactobacillus plantarum bacterial classification and enterococcus faecalis obtain lactobacillus acidophilus bacteria suspension, Lactobacillus plantarum bacteria suspension and enterococcus faecalis bacteria suspension.
Lactobacillus acidophilus culture medium prescription: peptone 9g, yeast extract 7g, whey powder 8g, beef extract 13 g, glucose 14 g, sodium acetate 4 g, sodium chloride 5g, ammonium citrate 2.4g, Tween-80 0.8ml, MgSO
47H
2o 0.63 g, MnSO
44H
2o 0.07 g, K
2hPO
42 g, agar powder 16g, water 1000 mL.
Lactobacillus plantarum culture medium prescription: yeast extract 7.5g, peptone 7.5g, whey powder 6g, beef extract 11g, glucose 10g, sodium chloride 5g, the KH2PO4 2g， 100ml of Tomato juice, Tween-80 0.5ml, distilled water 1000ml, pH 7.0.
Enterococcus faecalis culture medium prescription: peptone 9g, maltose 18g, yeast extract 5g, sodium glycero-phosphate 8g, sodium chloride 5g, lactose 0.8g, bromocresol purple 0.01g, Sodium azide 0.35g, TTC 0.1g, agar 15g
Distilled water 1000ml, pH value 7.2.
Respectively above-mentioned lactobacillus acidophilus bacteria suspension, Lactobacillus plantarum bacteria suspension and enterococcus faecalis bacteria suspension are filtered, and by the bacteria suspension of collecting after filtering by lactobacillus acidophilus bacterium: Lactobacillus plantarum bacterium: the part by weight of enterococcus faecalis bacteria suspension=1:1:2-2:2:1 mixes to obtain compound bacteria suspension, standby.
Step 2, the preparation of quick-release bacterium powder
The compound bacteria suspension of step 1 gained is filtered, in suspension after filtration, add account for filter after 8% to 12% beta-schardinger dextrin-of compound bacteria suspension gross weight, 5% to 8% porous-starch, soybean protein isolates 5%-8%, 13% to 16% sterilized water, adopts 900 revs/min of electric mixers to stir after 20-40 minute, adopt vacuum desiccator at vacuum condition to be-0.085MPa, temperature is dry 0.5h-1h at 25 ℃, mixes, and obtains quick-release bacterium powder 1.
The preparation of step 3, middling speed release type bacterium powder (the bacterium powder of solid wall material)
Under aseptic technique, take the porous-starch of 6g-10g, the shitosan of 6g-10g is in beaker, the sterilized water (take this 100mL sterilized water is follow-up interpolation material with reference to amount) that adds 100mL, the acetic acid (take 100mL sterilized water as reference) that adds 0.5wt%-0.8 wt %, adopt 1200 revs/min of electric mixers to stir after 20-40 minute, shitosan and porous-starch are dissolved completely, add the compound bacteria suspension 30g-40g being prepared by step 1, at room temperature adopt again supersonic generator concussion absorption 30-50 minute, add 2 wt % sodium alginate solns to obtain emulsion, with syringe, emulsion is clamp-oned to the Cacl that the concentration of 15-25mL is 2%
2in solution (aseptic), solidify 30min film forming.After filtering, by the concentration of 50-60mL, be 3.5%Cacl
2solution soaks 10min and hardens, and refilters and obtains sample, with clear water, floats surperficial Cacl
2raffinate, then use the physiological saline (0.85wt%) of sterilizing to wash surface of microcapsule thalline.Adopt the dry carbon dioxide shaft ice frozen 5-8 minute of-78 ℃ again, adopt vacuum desiccator at vacuum condition to be-0.085MPa, temperature is dry 0.5h-1h at 25 ℃, obtains middling speed release type bacterium powder (the bacterium powder of solid wall material) 2.
Step 4, the preparation of release type bacterium powder (the bacterium powder of double wall material) at a slow speed
Under aseptic technique, take 7g-11g zeins matter in beaker, add 100mL food-grade anhydrous ethanol (as follow-up additive with reference to amount), the bacterium powder of middling speed release type bacterium powder 25-35g(solid wall material prepared by step 3) be scattered in ethanol, adopt 1200 revs/min of electric mixers to stir after 20-40 minute, through low temperature spray drying, 90 ℃～105 ℃ of inlet temperatures, 60 ℃～65 ℃ of outlet temperatures, after spraying is dry, collects and obtain release type bacterium powder (the bacterium powder of double wall material) 3 at a slow speed.
The probiotics micro-ecological formulation mixed feed preparation of step 5, multistage controlled release
Respectively by quick-release bacterium powder 1, middling speed release type bacterium powder 2 and at a slow speed release type bacterium powder (the bacterium powder of double wall material) 3 mixes by 2:1:1 to 1:1:2 part by weight, get 100g Mixed Microbes powder (as follow-up additive with reference to amount), then add 7% to 9% bean dregs, 5% to 7% groundnut kernel powder, 3% to 5% WPC matter, 8 to 10% soybean protein isolates, dusty yeast 3% to 5%, fish oil 1.1% to 3.5%, corn flour 8.3 % to 10.5%, amino acid 2%, premix 1.3 % to 2.8%, starch 13 % to 15%, soy meal 10% to 13%, adopt 900 revs/min of electric mixers to stir after 20-40 minute, mix, obtain the probiotics micro-ecological formulation mixed feed of multistage controlled release.
The probiotics micro-ecological formulation mixed feed of the feeding multistage controlled release of this Silurus meridionalis Chen, when the fish such as cultivation Silurus meridionalis Chen, by 100 fishes, juvenile fish is raised 3-5 time every day, adult fish is raised 5-7 time every day, each 4kg-6kg, experimental result shows, this mixed feed can obviously promote the fish growths such as Silurus meridionalis Chen, raise after juvenile fish 45d, south catfish amount increase has improved 12.4% than control group respectively, protein efficiency ratio, PER improves 32.5%, the incidence of disease reduces 30%-40%, this probiotics mixed feed can improve the metabolism of body, supplement the nutritional labeling of body, beneficial bacterium can further be bred after entering alimentary canal, produce necessary nutriment in growth of animal process, improve the conversion ratio of feed, promote the growth of Silurus meridionalis Chen, and make the meat of Silurus meridionalis Chen more delicate, taste is more delicious, there is better edibility, bring up better economic worth.And a kind of probiotics preparation method for Silurus meridionalis Chen provided by the present invention, processing step is simple, and material therefor cost is low, is suitable for suitability for industrialized production, and range of application is wide.
Should be understood that, for those of ordinary skills, can be improved according to the above description or convert, and all these improvement and conversion all should belong to the protection domain of claims of the present invention.
Claims (1)
1. a preparation method for the probiotics micro-ecological formulation mixed feed of the feeding multistage controlled release of Silurus meridionalis Chen, is characterized in that, comprises the following steps:
Step 1, actication of culture and expansion are cultivated
After being activated respectively, expand cultivation, the lactobacillus acidophilus bacterial classification of refrigeration, Lactobacillus plantarum bacterial classification and enterococcus faecalis obtain lactobacillus acidophilus bacteria suspension, Lactobacillus plantarum bacteria suspension and enterococcus faecalis bacteria suspension; Respectively above-mentioned lactobacillus acidophilus bacteria suspension, Lactobacillus plantarum bacteria suspension and enterococcus faecalis bacteria suspension are filtered, and by the bacteria suspension of collecting after filtering by lactobacillus acidophilus bacterium: Lactobacillus plantarum bacterium: the part by weight of enterococcus faecalis bacteria suspension=1:1:2-2:2:1 mixes to obtain compound bacteria suspension, standby;
Step 2, the preparation of quick-release bacterium powder
The compound bacteria suspension of step 1 gained is filtered, in suspension after filtration, add account for filter after 8% to 12% beta-schardinger dextrin-of compound bacteria suspension gross weight, 5% to 8% porous-starch, soybean protein isolates 5%-8%, 13% to 16% sterilized water, adopts 900 revs/min of electric mixers to stir after 20-40 minute, adopt vacuum desiccator at vacuum condition to be-0.085MPa, temperature is dry 0.5h-1h at 25 ℃, mixes, and obtains quick-release bacterium powder 1;
The preparation of step 3, middling speed release type bacterium powder
Under aseptic technique, take the porous-starch of 6g-10g, the shitosan of 6g-10g is in beaker, the sterilized water that adds 100mL, take that this 100mL sterilized water is follow-up interpolation material with reference to amount, the acetic acid that adds 0.5wt%-0.8wt%, adopt 1200 revs/min of electric mixers to stir after 20-40 minute, shitosan and porous-starch are dissolved completely, add the compound bacteria suspension 30g-40g being prepared by step 1, at room temperature adopt again supersonic generator concussion absorption 30-50 minute, add 2wt% sodium alginate soln to obtain emulsion, with syringe, emulsion is clamp-oned to the CaCl that the concentration of 15-25mL is 2%
2in solution, solidify 30min film forming, after filtering, by the concentration of 50-60mL, be 3.5%Cacl
2solution soaks 10min and hardens, and refilters and obtains sample, with clear water, floats surperficial Cacl
2raffinate, then wash surface of microcapsule thalline with the physiological saline of sterilizing, then adopt the dry carbon dioxide shaft ice frozen 5-8 minute of-78 ℃, adopt vacuum desiccator at vacuum condition to be-0.085MPa, temperature is dry 0.5h-1h at 25 ℃, obtains middling speed release type bacterium powder 2,
Step 4, the preparation of release type bacterium powder at a slow speed
Under aseptic technique, take 7g-11g zeins matter in beaker, the food-grade anhydrous ethanol that adds 100mL, middling speed release type bacterium powder 25-35g prepared by step 3 is scattered in ethanol, adopts 1200 revs/min of electric mixers to stir after 20-40 minute, through low temperature spray drying, 90 ℃～105 ℃ of inlet temperatures, 60 ℃～65 ℃ of outlet temperatures, after spraying is dry, collects and obtain release type bacterium powder 3 at a slow speed;
The probiotics micro-ecological formulation mixed feed preparation of step 5, multistage controlled release
Respectively by quick-release bacterium powder 1, middling speed release type bacterium powder 2 and at a slow speed release type bacterium powder 3 mix by 2:1:1 to 1:1:2 part by weight, get Mixed Microbes powder, then add 7% to 9% bean dregs, 5% to 7% groundnut kernel powder, 3% to 5% WPC matter, 8% to 10% soybean protein isolates, dusty yeast 3% to 5%, fish oil 1.1% to 3.5%, corn flour 8.3% to 10.5%, amino acid 2%, premix 1.3% to 2.8%, starch 13% to 15%, soy meal 10% to 13%, adopt 900 revs/min of electric mixers to stir after 20-40 minute, mix, obtain the probiotics micro-ecological formulation mixed feed of multistage controlled release.
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| CN103283975A (en) * | 2013-03-27 | 2013-09-11 | 广州格拉姆生物科技有限公司 | A method of using spray drying for preparing probiotics micro-capsules |
| CN103525731A (en) * | 2013-10-15 | 2014-01-22 | 南京市水产科学研究所 | Slow-release microecologics and preparation method thereof |
| CN103981116A (en) * | 2013-12-24 | 2014-08-13 | 北京伟嘉人生物技术有限公司 | Culture method and application of enterococcus faecium |
| CN103981115B (en) * | 2013-12-24 | 2018-10-26 | 北京大伟嘉生物技术股份有限公司 | One plant height resistance enterococcus faecium and its application |
| ITUA20162345A1 (en) * | 2016-04-06 | 2017-10-06 | Nobile Matteo Alessandro Del | FISH PRODUCTS ENRICHED WITH PROBIOTICS AND ITS PRODUCTION PROCEDURE |
| CN113694086A (en) * | 2020-05-20 | 2021-11-26 | 香港理工大学深圳研究院 | Probiotics capsule and preparation method thereof |
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| CN1620892A (en) * | 2004-12-10 | 2005-06-01 | 中国海洋大学 | Compound microecological preparation for prawn |
| CN100581378C (en) * | 2006-08-01 | 2010-01-20 | 济南亿民动物药业有限公司 | Method for preparing fermented Chinese herbal medicine preparations for feed |
| CN100537759C (en) * | 2006-12-04 | 2009-09-09 | 济南赛拜斯生物工程有限公司 | Enteric-coated multilayer encapsulated probiotic microcapsule and preparation method thereof |
| CN101496555B (en) * | 2008-01-31 | 2011-09-21 | 北京大北农科技集团股份有限公司 | Lactobacillus micro-capsule as well as preparation method and use |
| CN101642189B (en) * | 2008-08-07 | 2012-05-30 | 北京大北农科技集团股份有限公司 | Microcapsule compound feed additive and preparation method thereof |
| CN101530163B (en) * | 2009-04-24 | 2011-08-24 | 孙瑞椒 | Method for keeping activity of probiotic |
| CN101991006A (en) * | 2009-08-13 | 2011-03-30 | 河南牧鹤(集团)饲料有限公司 | Silurus meridionalis feed and processing method thereof |
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